Ziconotide nasal-brain delivery preparation

Abstract

The invention provides a ziconotide nasal-brain delivery preparation which is high in brain targeting, safe and noninvasive and directly enters the brain through nasal administration. The nasal-brain delivery preparation comprises an active component ziconotide and an absorption enhancer, wherein the absorption enhancer is selected from one or more of dodecyl-beta-D-maltoside (DDM), menthol, hydroxypropyl-beta-cyclodextrin (HP-beta-CD), sodium glycocholate (SGC) and related derivatives thereof. The absorption enhancer with a specific type and content and the ziconotide cooperate and act together, so that the ziconotide bypasses a blood brain barrier, enters the brain through a nasal olfactory mucous membrane, forms a medicine storage bank in an olfactory bulb, is slowly released to the brain and keeps a long-term medicine effect, and therefore, a traditional ziconotide intrathecal administration mode is abandoned; the ziconotide nasal-brain administration mode is created, and the ziconotide nasal-brain administration method is a new-generation therapy for pain diseases related to the central nervous system.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to a pharmaceutical composition, particularly to a composition with high safety and high brain targeting, especially to a nasal-delivered formulation of ziconopeptide with high safety and high brain targeting. Background Technology

[0002] Pain is defined as an unpleasant sensory and emotional experience. Millions of people suffer from pain, ranging from a mild headache to debilitating chronic pain. Acute pain may be associated with medical procedures or injuries. Chronic pain may be associated with conditions such as cancer, HIV, or diabetes.

[0003] Ziconotide is an N-type calcium channel (NCC) blocker, a peptide toxin compound containing 25 amino acids and three disulfide bonds, extracted from the venom secreted by the South Pacific cone snail (Conus magus). Ziconotide is a polycationic peptide composed of 25 amino acids, including four lysines and two arginines as basic amino acids; its isoelectric point (pI) is 11.215, and its relative molecular mass is 2639.1. Ziconotide can relieve pain unresponsive to other treatments, including intrathecal morphine, and long-term use does not lead to tolerance or addiction. Its indications include the treatment of chronic pain associated with trauma, tumors, and neuropathic pain, with particular advantages in treating opioid-resistant pain or patients who cannot tolerate opioids.

[0004] However, ziconopeptide cannot cross the blood-brain barrier, requiring surgical implantation of a catheter inside the body for long-term intrathecal administration. This carries risks of infection, is expensive, and has other drawbacks. Clinically, there have been cases of infection at the microinjection site, uncontrollable bleeding, and spinal canal obstruction leading to impaired cerebrospinal fluid circulation. Long-term intrathecal administration may also cause severe psychotic symptoms and neurological damage. This method of administration significantly limits the clinical application of the drug's inherent advantages.

[0005] The blood-brain barrier (BBB) ​​is a complex cellular system located between brain tissue and blood. It controls the transport of substances across the blood-brain boundary, thus ensuring the stability of the central nervous system's internal environment. Useful substances in the blood interact with various receptors on the endothelial cell membranes of microvessels and are transported to the brain as needed to exert their effects. Conversely, toxic and harmful substances are shielded from brain damage by this barrier. This unique protective function of the BBB makes it difficult for most drugs to reach the brain, posing a challenge to the treatment and administration of drugs for central nervous system diseases.

[0006] Nasal administration is a non-invasive and convenient method. After absorption through the nasal mucosa, drugs can exert both local and systemic therapeutic effects. Unlike invasive procedures such as intravenous, intramuscular, and subcutaneous administration, patients can self-administer medication, resulting in better compliance. Drug absorption through the nasal mucosa occurs via the olfactory pathway, the trigeminal nerve pathway, and the bloodstream. The olfactory and trigeminal nerve pathways allow drugs to bypass the blood-brain barrier and directly enter the brain. In the olfactory pathway, drugs can be transported from olfactory nerve neuron axons to the olfactory bulb and then distributed to the brain parenchyma. Drugs can also be transported into the cerebrospinal fluid through the inter-cycle intervals of the olfactory nerve bundles. The bloodstream pathway allows drugs to bypass the first-pass effect of the liver, entering the bloodstream and then indirectly entering the brain via the blood-brain barrier. Therefore, nasal administration not only improves drug bioavailability but also increases drug release into the brain, giving it an advantage in treating brain diseases.

[0007] To facilitate ziconotide's entry into the brain, numerous studies have been conducted to improve it. Methods to promote the absorption of protein-peptide drugs through the nasal mucosa include: ① forming gels, such as a nasal gel composition disclosed in patent application 201310730052.X, which prolongs the drug's residence time in the nasal cavity by increasing the gel matrix, allowing ziconotide to enter the bloodstream. The drawback of this approach is that it does not directly enter the brain; ② chemically modifying peptide drugs, such as the method disclosed in patent application 201811086213.5, which involves linking a membrane-penetrating peptide to ziconotide to allow it to cross the blood-brain barrier. However, this substance is unstable and difficult to implement industrially; ③ Literature reports that delivery of ziconotide to cerebrospinal fluid via the intranasal pathway for the treatment of chronic pain involves adding chitosan to the ziconotide gel to prolong its action time in the nasal cavity, increasing the amount of drug entering the cerebrospinal fluid. However, this method does not achieve the slow release of ziconotide into the brain.

[0008] Therefore, obtaining a ziconovide nasobrain delivery drug formulation that can cross the blood-brain barrier, overcome the shortcomings of intrathecal administration, allow ziconovide to enter the brain directly via the nasal olfactory mucosa, and can be slowly released in the brain for large-scale clinical use is an urgent problem to be solved. Summary of the Invention

[0009] Through extensive experiments, the inventors discovered that by co-administering the specific absorption enhancer DDM with ziconovide to the olfactory region of the nasal cavity, DDM and ziconovide work synergistically to penetrate the olfactory mucosa and enter the olfactory bulb, forming a drug reservoir within the bulb and slowly releasing it to the brain. This can treat pain symptoms related to the central nervous system. Researchers found that by controlling the appropriate percentage content of DDM and ziconovide, a ziconovide nasobrain delivery formulation can be obtained that is non-ciliary toxic, can be administered non-invasively, can rapidly enter the brain, forms a drug reservoir in the olfactory bulb, and can be slowly released into the brain.

[0010] The objective of this invention is achieved through the following technical solutions: In one aspect, the present invention provides a ziconopeptide nasal-brain delivery formulation comprising the active ingredient ziconopeptide and its pharmaceutically acceptable salts and absorption enhancers, and water as a solvent.

[0011] In some embodiments, the absorption enhancer is selected from alkyl glycosides, such as decaalkyl-β-D-maltodextrin, dodecyl-β-D-maltodextrin, tetradecyl-β-D-maltodextrin, and dodecyl glucoside; or fatty acid salts, such as sodium glycocholate, sodium heptanoate, sodium taurocholate, sodium caprylate, sodium dodecyl sulfate, sodium 8-(2-hydroxybenzamido)caprylate, disodium edetate, and sodium docusate; or cyclodextrin derivatives, such as hydroxypropyl-α-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, methyl-beta-cyclodextrin, and cationic-β-cyclodextrin. Sulfobutyl-β-cyclodextrin; or nonionic surfactants, such as polyethylene glycol 1000 vitamin E succinate, poloxamer, polysorbate 80, polyoxyethylene hydrogenated castor oil (RH40), sucrose dodecanoate, polyethylene glycol 15-hydroxystearate, polyethylene glycol glycerol caprylate; or at least one of menthol, chitosan, decyl-L-carnitine, dodecyl phosphate choline, and cyclopentadecanol; preferably at least one of menthol, polysorbate 80, dodecyl-β-D-maltodextrin, and sodium glycocholate, most preferably dodecyl-β-D-maltodextrin.

[0012] In some embodiments, the absorption enhancer is present in a content of 0.01% to 4% by weight of the total weight of the nasoencephalometric preparation, preferably 0.05% to 2.0%, and non-limiting embodiments include: 0.10%, 0.20%, 0.30%, 0.40%, 0.50%, 1.0%, 1.5%, 2.0%, or any value between any two of these.

[0013] In some embodiments, the active ingredient, by weight, accounts for 0.01% to 1% of the total weight of the nasoencephalogenic formulation, preferably 0.05% to 0.5%, and non-limiting embodiments include: 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, or any value between any two.

[0014] Furthermore, the ziconopeptide nasal-brain delivery formulation of the present invention may also include an osmotic pressure regulator and an antioxidant.

[0015] In some embodiments, the osmotic pressure regulator is selected from sodium chloride, glucose, potassium chloride, mannitol, sorbitol, sucrose, dextran, trehalose, and preferably sodium chloride.

[0016] In some embodiments, the osmotic pressure regulator, by weight, accounts for 0.01-10% of the total weight of the nasocerebrospinal release solution, preferably 0.1%-5%, and non-limiting embodiments include 0.1%, 0.5%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.5% or any value between any two, particularly preferably 0.9%.

[0017] In some embodiments, the antioxidant is selected from methionine, glutathione, tyrosine, tryptophan, valine, lipoic acid, butylated hydroxyanisole, 2,6-di-tert-butyl-p-cresol, vitamin C, sodium bisulfite, butylated hydroxyanisole, and preferably, methionine.

[0018] In some embodiments, the antioxidant, by weight, accounts for 0.01%-5% of the total weight of nasoencephalogenic release, preferably 0.05%-2%, and non-limiting embodiments include 0.05%, 0.10%, 0.15%, 0.16%, 0.2%, 0.3%, 0.4%, 0.5%, 1.0%, 1.5%, 2.0%, or any value between any two, particularly preferably 0.16%.

[0019] In some embodiments, the ziconopeptide nasal-brain delivery formulation of the present invention further includes an antibacterial agent selected from benzalkonium chloride, potassium sorbate, phenoxyethanol, chlorobutanol, benzyl alcohol, methylparaben, and propylparaben, preferably benzalkonium chloride.

[0020] In some embodiments, the antibacterial agent, by weight, accounts for 0.0001%-0.5% of the total weight of the nasoencephalogenic formulation, preferably 0.001%-0.1%, and non-limiting embodiments include 0.005%, 0.010%, 0.015%, 0.02%, 0.025%, 0.03%, 0.04%, 0.05%, or any value between any two, particularly preferably 0.01%.

[0021] The ziconopeptide nasal-brain delivery formulation of the present invention is in solution form.

[0022] In some specific embodiments, the pH value of the ziconopeptide nasal-brain delivery formulation is 4.0 to 5.0.

[0023] In some specific embodiments, the ziconovide nasobrain delivery formulation comprises 0.05% to 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.05% to 2.0% dodecyl-β-D-maltodextrin, 0.1% to 5% sodium chloride, and 0.05% to 2% methionine; optionally, it further comprises 0.001% to 0.1% benzalkonium chloride, and the pH value of the ziconovide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconovide nasobrain delivery formulation comprises 0.05% to 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconopeptide nasobrain delivery formulation comprises 0.01% ziconopeptide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconopeptide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconovide nasobrain delivery formulation comprises 0.05% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconovide nasobrain delivery formulation comprises 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.0.

[0024] In some embodiments of the present invention, the ziconopeptide nasobrain delivery formulation contains the above-mentioned ziconopeptide, the above-mentioned absorption enhancer, the above-mentioned osmotic pressure regulator, the above-mentioned antioxidant, and water as a solvent, and optionally, also contains the above-mentioned antibacterial agent.

[0025] In certain specific embodiments of the present invention, a high concentration of the ziconopeptide nasobrain delivery formulation of the present invention in the olfactory region can serve as a drug reservoir, maintaining a relatively stable concentration in the brain for a long time, thereby exerting a longer-lasting therapeutic effect.

[0026] According to a particularly preferred embodiment, the ziconopeptide nasal-brain delivery formulation of the present invention comprises the above-mentioned ziconopeptide, the above-mentioned absorption enhancer, the above-mentioned osmotic pressure regulator, the above-mentioned antioxidant, and water as a solvent.

[0027] According to another particularly preferred embodiment, the ziconopeptide nasobrain delivery formulation of the present invention is delivered via the small animal intelligent integrated nasal drug delivery system (RA-IDDS) developed by the applicant company. ® Dosage, as disclosed in CN202021777601.0 and CN202010148279.3.

[0028] A second aspect of the present invention provides a method (I) for preparing a ziconopeptide nasal-brain delivery formulation, the main steps of which include: Ziconopeptide, the above-mentioned osmotic pressure regulator, and the above-mentioned antioxidant are dissolved in water to prepare a solution without absorption promoter; Adjust the pH value to 4.0~5.0; Add the absorption enhancer in the prescribed proportion and stir to dissolve; In one specific embodiment, a method (II) for preparing a ziconopeptide nasal-brain delivery formulation is provided, the main steps of which include: 1) Weigh N times the prescribed amount of ziconopeptide, the prescribed amount of osmotic pressure regulator, antioxidant, and the prescribed amount of water to prepare a high-concentration ziconopeptide solution; 2) Weigh the prescribed amount of osmotic pressure regulator, antioxidant, and water, and prepare a blank solution without active ingredients; 3) Take a certain proportion of high-concentration ziconopeptide solution and blank solution without active ingredients, and mix them to form the ziconopeptide nasal and brain delivery preparation in the prescription ratio; Optionally, the preparation methods (a) and (b) further include the step of dissolving the antibacterial agent into the ziconopeptide nasal-brain delivery formulation.

[0029] A fourth aspect of the present invention provides the use of the above-mentioned ziconopeptide naso-brain delivery formulation in the preparation of a pain medication.

[0030] In one specific embodiment, the pain is selected from the following types: severe chronic and intractable pain such as postherpetic neuralgia, trigeminal neuralgia, diabetic peripheral neuropathy, HIV infection pain, headache, limb pain, and joint pain caused by multiple sclerosis; traumatic mechanical injury pain such as compression neuropathy, nerve transection injury, burns, and spinal cord injury; metabolic or nutritional pain such as injury pain, postoperative pain, alcoholic neuropathy pain, and phantom limb pain; cancer pain and pain caused by radiotherapy and chemotherapy; pain caused by heavy metal poisoning such as thallium and arsenic; and pain caused by neurotransmitter disorders such as thalamic syndrome, stroke, and neurodegenerative diseases.

[0031] A fifth aspect of the present invention provides the application of the above-mentioned ziconopeptide naso-brain delivery formulation in the preparation of analgesics via nasal delivery to the brain, wherein the nasal delivery to the brain is a micro-delivery to a precise nasal olfactory region at a constant rate or in a pulsed manner; or the nasal delivery to the brain is a delivery to the nasal olfactory region via targeted spraying.

[0032] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0033] Compared with the prior art, the present invention has the following beneficial effects: 1. The nasobrain delivery formulation of this patent application can form a drug reservoir in the olfactory bulb, allowing ziconotide to be slowly released in the brain over a period of up to 2 hours, thus reducing the frequency of administration.

[0034] 2. The naso-brain delivery formulation of this patent application, after being administered via the nasal olfactory region, exhibits high selectivity for brain entry, with the Cmax ratio of ziconopeptide entering the olfactory bulb to entering the bloodstream reaching as high as 11.3.

[0035] 3. The nasobrain delivery formulation of this patent application has a significant analgesic effect 10 minutes after administration through the nasal olfactory region, and no special behaviors related to central adverse reactions of ziconopeptide, such as leg retraction, tail tremor, or whole-body tremor, were observed after administration. In other words, it has better safety compared with intrathecal administration.

[0036] 4. The nasobrain delivery formulation of this patent application can be administered via the nasal olfactory region, resulting in better subject compliance compared to intrathecal injection. Attached Figure Description

[0037] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings: Figure 1 This is the pharmacokinetic diagram of the ziconopeptide nasal-brain delivery formulation of pharmacological example 1 after nasal administration; Figure 2This is a graph showing the therapeutic effects of different doses of the ziconopeptide nasal-brain delivery formulation administered via nasal route in Pharmacological Example 2 on pain. Figure 3 This is pharmacological example 3, the test results of the analgesic effect of the drug on the SD rat foot incision pain model (postoperative analgesia) (pain threshold change value). Figure 4 This is the analgesic effect test result (pain threshold increase rate) of the drug on the SD rat foot incision pain model (postoperative analgesia) in pharmacological example 3. Figure 5 It is the structure of impurity A, impurity B, impurity C, and impurity D. Detailed Implementation

[0038] The measurement method of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0039] Terminology Explanation: DDM refers to dodecyl-β-D-maltodextrin; P188 refers to Polosham 188; SGC refers to sodium glycocholate. M-β-CD refers to methyl-β-cyclodextrin; HP-β-CD refers to hydroxypropyl-β-cyclodextrin; BHT refers to 2,6-di-tert-butyl-p-cresol; API refers to Ziconopeptide.

[0040] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0041] In this patent application, the absorption rate constant refers to the absorption rate constant of rat nasal perfusion. The determination method employs methods known to those skilled in the art. The method for determining rat nasal perfusion in this invention is as follows: a rat respiratory pathway is established through tracheal intubation; then, an esophageal cannula is inserted, and a peristaltic pump provides the driving force for drug circulation, establishing an in vivo nasal circulation pathway. The drug flows through the esophageal cannula across the nasal mucosa, returns to a constant-temperature collection bottle, and then enters the next cycle via the peristaltic pump. In the rat nasal circulation pathway, the drug is partially absorbed through the nasal mucosa. The remaining drug content in the collection bottle of the circulation system is determined by HPLC or other detection methods, and the amount of drug absorbed by the rat nasal mucosa is calculated. In this patent application, the absorption rate constant is determined using three rats, and the average value is calculated. (Reference: 1) Huang CH, Kimura R, Nassar RB, Hussain A. Mechanism of nasal absorption of drugs I:Physicochemical parameters influencing the rate of in situ nasal absorption of drugs in rats. J Pharm Sci. 1985 Jun;74(6):608-11.

[0042] In this invention, the local irritation test of the nasal cavity is determined using a ciliary toxicity model, which is a method known to those skilled in the art: the palatal cilia of toads are widely used as a commonly used model to simulate human nasal cilia [1]. After the test drug is dropped onto the mucosal surface for 60 min, the palatal mucosa is washed and separated. The continuous movement of the mucosal cilia is observed under an optical microscope, and its effect on the cilia is analyzed by comparing it with physiological saline as a control [2]. The movement of the cilia is observed, and the time required from the start of drug administration to the complete cessation of cilia movement is recorded, i.e., the continuous movement time of the cilia. The continuous movement time of the palatal mucosal cilia of the drug administration group is divided by the continuous movement time of the palatal mucosal cilia of the negative control group to obtain the relative percentage of the continuous movement time of the cilia. The higher this percentage, the less toxic the preparation is to the cilia, and the higher the safety of the preparation. The relative percentage of the continuous movement time of the cilia is greater than 85% as the threshold for the safety of the preparation. ([1] Jiang Xinguo, Cui Jingbin. Nasal mucociliary toxicity of drugs and evaluation methods[J]. Acta Pharmaceutica Sinica, 1995, 30:848-53. [2] Li Xinfang, Li Xiangui, Ma Zhiqiang, et al. Preparation and safety study of menthol nasal in situ gel[J]. Journal of Pharmaceutical Practice, 2017, 3(4):321-4.) Example 1: The transnasal absorption of teicoplanin was investigated using different types and amounts of absorption enhancers. The formulations are shown in Table 1 below. Table 1

[0043] Preparation method: Dissolve water, methionine, sodium chloride and ziconopeptide in the proportions of each prescription in Table 1 by stirring. Adjust the pH value to about 4.3 by adding a certain concentration of hydrochloric acid solution to prepare a solution without absorption promoter. Then add the absorption promoter in the proportion of the prescription and stir to dissolve.

[0044] The pH value, osmotic pressure, and transnasal mucosal absorption rate constant of each prescription sample were tested, and the results are shown in Table 2.

[0045] Table 2

[0046] Experimental conclusion: The absorption-enhancing effect of 0.2% DDM was excellent, and even when the DDM concentration was reduced from 0.2% to 0.1%, the absorption-enhancing effect remained excellent. 2% HP-β-CD and menthol showed moderate effects on the transmembrane absorption of ziconopeptide, while 1% P188 showed even worse absorption-enhancing effects. The nasal mucosal absorption rate constants of 2% Tween 80 and 2% menthol were lower than the detectable lower limit of ziconopeptide absorption in a rat nasal perfusion model. ND indicates absorption below the lower limit of detection. When SGC was used as the absorption enhancer, 2% SGC showed good absorption-enhancing effects, but adverse reactions occurred in rats during perfusion, releasing excessive protein into the perfusion fluid, making it unsuitable as an absorption enhancer for nasoencephalogenic formulations.

[0047] Example 2: Nasal mucosa irritation test The formulations listed in Table 3 below are prepared using methods conventional to those skilled in the art, or according to the following method. Dissolve water, methionine, sodium chloride, 10 times the amount of ziconovide, and 0.2% DDM (or without DDM) in the formulation proportions shown in Table 3 by stirring. Adjust the pH to approximately 4.3 by adding a certain concentration of hydrochloric acid solution dropwise to prepare a high-concentration ziconovide solution. Weigh out the methionine, sodium chloride, and DDM (or without DDM) in the formulation proportions and dissolve them in the formulation proportions of water to form a blank solution without ziconovide. Take an appropriate proportion of the high-concentration ziconovide and the blank solution, and mix them to obtain the ziconovide nasal-brain delivery formulation at the formulation concentration. For example, taking the preparation method of prescription 2-4 as an example, weigh 0.5 mg of ziconovide, 0.16 mg of methionine, 0.85 mg of sodium chloride, and 0.05 mg of DDM and dissolve them in 97.44 mg of water to form a high-concentration ziconovide solution; then weigh 0.16 mg of methionine, 0.85 mg of sodium chloride, and 0.05 mg of DDM and dissolve them in 98.94 mg of water to form a blank solution without ziconovide; mix 1 part by weight of the concentrated ziconovide solution and 9 parts by weight of the blank solution evenly to obtain the nasoencephalogenic formulation of ziconovide of prescription 2-4.

[0048] Table 3

[0049] The pH value, osmotic pressure, and nasal mucosa and ciliary toxicity of each prescription sample were tested, and the results are shown in Table 4.

[0050] Table 4

[0051] Experimental conclusion: Ziconovide nasal-brain delivery solution containing a high concentration of DDM is toxic to the nasal mucosa and cilia and is not suitable as a nasal-brain delivery solution; when the DDM content is reduced to 0.1%, the nasal mucosa and cilia toxicity of the prescriptions with ziconovide concentrations of 0.5 mg / g and 5 mg / g both meet the standards.

[0052] Example 3: Determination and Selection of pH Value Solutions with different pH values ​​were prepared as shown in Table 5 below. The preparation methods were similar to those in Example 1 or other known methods, and are not specifically limited herein. The specific structures of the impurities are shown in the attached table. Figure 5 As shown, impurity A and impurity B are isomers of each other. Although they have the same structural formula, they have different peak positions in the analysis and detection.

[0053] Table 5

[0054] The preparation process was carried out without nitrogen purging, and the solution was stored at 40℃±2℃ in the dark. Samples were taken at 7 and 14 days to test the product's properties (clarity and color of the solution, visible foreign matter), pH value, and related substances. The effect of pH value on product quality was investigated. The results are shown in Table 6 below. Table 6

[0055] Note: Only impurities ≥0.05 are statistically calculated. Experimental conclusion: When a sample with a pH of 3.0 was placed at 40°C for 14 days, visible foreign matter was found, and the related substance impurity C increased significantly to 1.1%, which did not meet the quality standard. When a sample with a pH of 6.0 was placed at 40°C for 14 days, the levels of related substance impurities (D), unknown impurities, and total impurities increased significantly, while the sample content decreased significantly; thus, it did not meet the quality standards. Therefore, the pH range of the product is determined to be 4.0~5.0.

[0056] Example 4: Determination of Methionine Dosage The effect of different amounts of methionine on product quality was investigated. Solutions as shown in Table 7 were prepared according to a method similar to that described in Example 1 or according to methods known to those skilled in the art; no limitation is made herein. Table 7

[0057] Nitrogen was not added during the preparation process and after the preparation was completed.

[0058] Prepare each formulation according to the table above, store at 40℃±2℃ in the dark, and take samples at 7 and 14 days to test its properties (clarity and color of the solution, visible foreign matter), pH value, and related substances. The results are shown in Table 8 below: Table 8

[0059] Experimental conclusion: For samples with methionine concentrations between 0.005% and 0.16%, when placed at 40℃, the 0.005% methionine solution (Formula 8) began to show foreign matter after 7 days, proving that the sample was unqualified under the condition of 0.005% methionine. This also proves that a methionine content range of 0.08% to 0.16% contributes to product stability.

[0060] Example 5: Investigation of residual oxygen content The effect of different residual oxygen levels on product quality was investigated. The preparation process can follow a method similar to that in Example 1 or a method conventional to those skilled in the art. The formulations shown in Table 9 below were prepared. Table 9

[0061] Note: Residual oxygen volume (ml) = Residual oxygen % * Packaging material headspace volume. 1ml ampoule headspace volume is 1.33ml, 2ml vial headspace volume is 3.21ml.

[0062] The sample with a residual oxygen content of 21% was unpurified with nitrogen, while the sample with a residual oxygen content of 1.07% was prepared by filling vials into lyophilized bottles, cyclically vacuuming and purging nitrogen in a freeze dryer, and then sealing and capping. Each formulation was prepared according to the table above, with formulations 5-4 tested in both nitrogen-purified and unpurified forms. Samples were stored at 40℃±2℃ in the dark, and taken at 7 and 14 days to analyze properties (clarity and color of the solution, visible foreign matter), pH value, and related substances (at 14 days). The experimental results are shown in Table 10 below. Table 10

[0063]

[0064] Experimental conclusion: It was unexpectedly discovered that, under this formulation, nitrogen purging can be omitted to maintain the stability of the formulation; while ziconopeptide intrathecal injection and PRIALT intrathecal injection require nitrogen purging to maintain the stability of the drug solution.

[0065] Pharmacological Example 1: The ziconopeptide nasal-brain delivery solution prepared according to prescriptions 2-4 is placed in a nasal delivery device (such as the RA-IDDS developed by the company that filed this patent application). ® The small animal intelligent integrated nasal drug delivery system (the drug delivery method and equipment can refer to the technology disclosed in patent applications CN202021777601.0 and CN202010148279.3) administers a prescribed dose of drug to rats via the nose. The nasal delivery method involves a constant rate of administration over 2 minutes or a pulsed micro-delivery to the precise olfactory region. Plasma samples were collected at 5, 10, 30, 60, and 120 minutes after administration to evaluate the pharmacokinetics and distribution of the nasal delivery formulation. Olfactory bulb and cerebral cortex samples were collected from the administration side at 2, 5, 10, 30, and 60 minutes after administration. The sampling methods for plasma, olfactory bulb, and cerebral cortex were employed using methods commonly used by those skilled in the art. Experimental results are shown in Table 11 below. Figure 1 As shown, Table 11

[0066] Experimental conclusion: The naso-brain delivery formulation of this patent application exhibits high brain targeting. The Cmax ratio of ziconopeptide entering the olfactory bulb to entering the bloodstream is as high as 11.3. Figure 1It is understood that the ziconovide naso-brain delivery solution of this patent application can form a drug reservoir in the olfactory bulb and enable ziconovide to be slowly released in the brain to exert its therapeutic effect, with a release time of up to 2 hours. Therefore, it can reduce the frequency of administration and improve patient compliance.

[0067] Pharmacological Example 2: Mouse Acetic Acid Writhing Test The analgesic effect of the ziconovide naso-brain delivery formulation was evaluated using the acetic acid writhing test; in this embodiment, IN refers to the nasal administration method.

[0068] The model group refers to the blank group, which refers to the ziconopeptide nasal-brain delivery formulation of this patent application that was not administered to mice before or after the injection of acetic acid. After administering the nasocerebrospinal release solution of prescriptions 2-4 of this application via the nasal route for 10 minutes, acetic acid was injected. The experimental results are shown in Table 12 below. Figure 2 As shown: Table 12

[0069] Where writhing inhibition rate % = (mean writhing in model group - mean writhing in treatment group) / mean writhing in model group * 100%; *p < 0.05, **p < 0.01, ***p < 0.001, compared with the model group.

[0070] Experimental conclusion: The ziconopeptide nasocephalic release solution of this patent application exhibits a good dose-response relationship within the dosage range of 0.0125~0.2 mg / kg. At a dose of 0.05~0.2 mg / kg, mice showed significant analgesic effects 10 minutes after administration, and no specific behaviors associated with central nervous system adverse reactions of ziconopeptide, such as leg retraction, tail tremor, or generalized tremor, were observed after administration, indicating superior safety compared to intrathecal administration.

[0071] The intrathecal reference drug PRIALT has a rat efficacy dose of 0.0012 mg / kg, while in rat safety pharmacology experiments, intrathecal injection of 0.0004 mg / kg resulted in central nervous system adverse reactions such as leg retraction, tail tremor, and generalized tremor.

[0072] This demonstrates that the formulation in this patent application, which uses naso-cerebrospinal administration, has a larger dose safety window and better efficacy.

[0073] Pharmacological Example 3: Evaluation of the analgesic effect of ziconopeptide nasal-brain delivery formulation on postoperative pain Experimental methods: The plantar incision model was used to evaluate the analgesic effect of ziconopeptide nasal-brain delivery on postoperative pain.

[0074] Pain threshold improvement rate % = (average mechanical foot retraction threshold in the treatment group - average mechanical foot retraction threshold in the model group) / average mechanical foot retraction threshold in the model group * 100%. Foot retraction threshold and other data are obtained through observation.

[0075] In this embodiment, the positive control drug refers to pregabalin, ip refers to intraperitoneal injection, and IN refers to nasoencephalogenic administration; the model group refers to the saline group; and the experimental group is the formulation of this patent application, for example, the nasoencephalogenic formulation of formulations 2-4 can be selected.

[0076] The experimental results are shown in Tables 13 and 14 below. Figure 3 and Figure 4 As shown: Table 13

[0077] Table 14

[0078] Experimental conclusion: In rats, intranasal administration of 0.01–0.1 mg / kg showed analgesic effects, peaking at 4 hours, with a good dose-response relationship. The high-dose group (0.1 mg / kg) demonstrated superior analgesic efficacy and duration of action over 4–8 hours compared to the positive control group (pregabalin 30 mg / kg), and maintained significant postoperative analgesia even after 8 hours. The medium-dose group (0.05 mg / kg) showed comparable analgesic efficacy and duration of action over 1–8 hours compared to the positive control group (pregabalin 15 mg / kg), and maintained significant postoperative analgesia even after 8 hours. Theoretically, the lower limb pain pathway ascends to the central nervous system, branching extensively; blocking pain requires all relevant brain regions to reach an effective concentration. However, the rat tissue distribution results in Pharmacological Example 1 showed that the ziconopeptide nasal-brain delivery formulation of this patent application distributed slowly along the olfactory nerve pathway to the cerebral cortex, with a stable concentration in the cerebral cortex, remaining in a plateau phase at 120 minutes. The PK-PD results showed good correlation.

[0079] The above-described embodiments are merely examples of several implementation methods of this disclosure. They are described in a relatively specific and detailed manner, but they should not be construed as limiting the scope of this patent disclosure. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of this disclosure, and these all fall within the protection scope of this disclosure.

Claims

1. A ziconopeptide nasal-brain delivery formulation, comprising: The active ingredient, ziconopeptide or its pharmaceutically acceptable salt, absorption enhancer, osmotic pressure regulator, antioxidant, and The solvent is water, and the pH value of the ziconopeptide nasal-brain delivery formulation is 4.0-5.0; The absorption enhancer mentioned above is dodecyl-β-D-maltodextrin; The content of the active ingredient, by weight, is 0.01% to 1% of the total weight of the nasocerebrospinal delivery formulation; The content of the absorption enhancer, by weight, is 0.01% to 4% of the total weight of the nasocerebrospinal delivery formulation; The osmotic pressure regulator is selected from one of sodium chloride, glucose, potassium chloride, mannitol, sorbitol, sucrose, dextran, and trehalose; The antioxidant is selected from one of the following: methionine, glutathione, tyrosine, tryptophan, valine, lipoic acid, butylated hydroxyanisole, 2,6-di-tert-butyl-p-cresol, vitamin C, sodium bisulfite, and butylated hydroxyanisole.

2. The ziconopeptide nasal-brain delivery formulation according to claim 1, characterized in that, The content of the osmotic pressure regulator, by weight, accounts for 0.01-10% of the total weight of the nasocerebrospinal delivery formulation; The antioxidant content, by weight, accounts for 0.01%-5% of the total weight of nasoencephalogenic release.

3. The ziconopeptide nasal-brain delivery formulation according to claim 1, characterized in that, The content of the active ingredient, by weight, is 0.05% to 0.5% of the total weight of the nasocerebrospinal delivery formulation; and / or, The absorption enhancer, by weight, accounts for 0.05% to 2.0% of the total weight of the nasocerebrospinal delivery formulation; and / or, The content of the osmotic pressure regulator, by weight, is 0.1%-5% of the total weight of the nasocerebrospinal delivery formulation; and / or, The antioxidant content, by weight, accounts for 0.05%-2% of the total weight of nasoencephalogenic release.

4. The ziconopeptide nasal-brain delivery formulation according to claim 1, characterized in that, The content of the active ingredient, by weight, is 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50% or any value between any two of the total weight of the naso-cerebrospinal delivery formulation; and / or, The absorption enhancer, by weight, comprises 0.10%, 0.20%, 0.30%, 0.40%, 0.50%, 1.0%, 1.5%, 2.0% of the total weight of the nasocerebrospinal delivery formulation, or any value between any two of these values; and / or, The content of the osmotic pressure regulator, by weight, is 0.1%, 0.5%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.5%, or any value between any two of the total weight of the nasocerebrospinal delivery formulation; and / or, The antioxidant content, by weight, is 0.05%, 0.10%, 0.15%, 0.16%, 0.2%, 0.3%, 0.4%, 0.5%, 1.0%, 1.5%, 2.0% or any value between any two of the total weight of the nasoencephalogenic release.

5. The ziconopeptide nasal-brain delivery formulation according to claim 1, characterized in that, The osmotic pressure regulator is sodium chloride, and the content of the osmotic pressure regulator, by weight, accounts for 0.9% of the total weight of the nasocerebrospinal delivery formulation; and / or, The antioxidant is methionine, and the content of the antioxidant, by weight, accounts for 0.16% of the total weight of the nasoencephalometric preparation.

6. The ziconopeptide nasal-brain delivery formulation according to claim 1, characterized in that, The ziconopeptide nasal-brain delivery formulation optionally further includes an antibacterial agent selected from one of benzalkonium chloride, potassium sorbate, phenoxyethanol, chlorobutanol, benzyl alcohol, methylparaben, and propylparaben; preferably, it is benzalkonium chloride. And / or, the content of the antibacterial agent, by weight, is 0.0001%-0.5% of the total weight of the ziconopeptide nasal-brain delivery formulation, preferably 0.001%-0.1%, for example 0.005%, 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.04%, 0.05% or any value between any two, particularly preferably 0.01%.

7. The ziconopeptide nasal-brain delivery formulation according to claim 6, characterized in that, The ziconovide naso-brain delivery formulation comprises 0.05% to 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.05% to 2.0% dodecyl-β-D-maltodextrin, 0.1% to 5% sodium chloride, and 0.05% to 2% methionine; optionally, it further comprises 0.001% to 0.1% benzalkonium chloride, and the pH of the ziconovide naso-brain delivery formulation is 4.0 to 5.

0. Alternatively, the ziconovide nasobrain delivery formulation comprises 0.05% to 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconopeptide nasobrain delivery formulation comprises 0.01% ziconopeptide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconopeptide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconovide nasobrain delivery formulation comprises 0.05% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.0; Alternatively, the ziconovide nasobrain delivery formulation comprises 0.5% ziconovide or a pharmaceutically acceptable salt thereof, 0.1% dodecyl-β-D-maltodextrin, 0.9% sodium chloride, 0.16% methionine, and optionally, 0.01% benzalkonium chloride, wherein the pH of the ziconovide nasobrain delivery formulation is 4.0 to 5.

0.

8. A method for preparing a ziconopeptide nasal-brain delivery formulation, comprising preparation method (I) or preparation method (II), characterized in that, The main steps of the preparation method (I) include: 1) Dissolve the ziconopeptide, the osmotic pressure regulator, and the antioxidant in the ziconopeptide nasal and brain-delivery formulation according to any one of claims 1 to 5 in water to prepare a solution without an absorption promoter. 2) Adjust the pH value to 4.0~5.0; 3) Add the absorption enhancer in the ziconopeptide nasal-brain delivery formulation according to any one of claims 1 to 5 in the prescribed proportion, and stir to dissolve; Alternatively, the main steps of the preparation method (II) include: 1) Weigh N times the amount of ziconovide in the ziconovide nasal-brain delivery formulation according to any one of claims 1 to 5, the amount of osmotic pressure regulator in the ziconovide nasal-brain delivery formulation according to any one of claims 1 to 5, the amount of antioxidant in the ziconovide nasal-brain delivery formulation according to any one of claims 1 to 5, and the amount of water according to the prescription, and prepare a high-concentration ziconovide solution. 2) Weigh the osmotic pressure regulator, the antioxidant, and the water in the ziconopeptide nasal and brain-delivery formulation according to any one of claims 1 to 5, and prepare a blank solution without active ingredients. 3) Take a certain proportion of high-concentration ziconopeptide solution and blank solution without active ingredients, and mix them to form the ziconopeptide nasal and brain delivery preparation in the prescription ratio; Optionally, the preparation method (a) or preparation method (b) further includes the step of dissolving the antibacterial agent in the ziconopeptide nasal-brain delivery formulation according to claim 6 into the ziconopeptide nasal-brain delivery formulation.

9. A ziconopeptide nasal-brain delivery formulation prepared by the method for preparing a ziconopeptide nasal-brain delivery formulation according to claim 8; preferably, the ziconopeptide nasal-brain delivery formulation does not require nitrogen filling.

10. Use of the ziconopeptide nasobrain delivery formulation according to any one of claims 1 to 7, 9 in the preparation of a pain medication; preferably, the pain is selected from the following types: severe chronic and intractable pain such as postherpetic neuralgia, trigeminal neuralgia, diabetic peripheral neuropathy, HIV infection pain, headache, limb pain, and joint pain caused by multiple sclerosis; traumatic mechanical injury pain such as compression neuropathy, nerve transection injury, burns, and spinal cord injury; metabolic or nutritional pain such as injury pain, postoperative pain, alcoholic neuropathy pain, and phantom limb pain; cancer pain and pain caused by radiotherapy and chemotherapy; pain caused by heavy metal poisoning such as thallium and arsenic; pain caused by neurotransmitter disorders such as thalamic syndrome, stroke, and neurodegenerative diseases; more preferably, the ziconopeptide nasobrain delivery formulation is administered via nasal delivery to the brain, wherein the nasal delivery to the brain is a micro-delivery to the precise olfactory region at a constant rate or in a pulsed manner, or the nasal delivery to the brain is a targeted spray delivery to the olfactory region.

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