A lyophilized dexamethasone sodium phosphate composition for tympanic membrane administration and a method for preparing the same

Abstract

The present application belongs to the field of biological medicine, and relates to a dexamethasone sodium phosphate lyophilized composition suitable for tympanic administration and a preparation method thereof. The present application provides a lyophilized powder injection preparation, which is a dexamethasone lyophilized powder injection, does not contain a lyophilized support agent or an excipient or a support agent, and contains a pharmaceutically acceptable auxiliary material glycerol, wherein the glycerol is added before lyophilization of the lyophilized powder injection preparation. The lyophilized preparation provided by the present application does not contain a lyophilized support agent or an excipient, and only a small amount of glycerol is added before lyophilization of the preparation to achieve the effect of improving the stability of the drug. Meanwhile, the osmotic pressure can be adjusted to be closer to the physiological osmotic pressure during tympanic administration, the local administration time is prolonged, the safety and effectiveness of tympanic administration are increased.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to a dexamethasone sodium phosphate lyophilized composition suitable for tympanic membrane administration and its preparation method. Background Technology

[0002] In recent years, glucocorticoids have been widely used to treat various inner ear diseases, such as deafness, Meniere's disease, and post-meningolabyrinthitis. Sudden deafness is a sudden, non-fluctuating sensorineural hearing loss with rapid onset and progression, causing a sudden decline in hearing within minutes, hours, or three days. Common accompanying symptoms include tinnitus and vertigo. The exact cause of sudden deafness is not fully understood, but the main causes include inner ear microcirculatory disorders, specific infections, immune factors, and psychological factors. Acute deafness has similar but different causes to sudden deafness. Acute deafness is accompanied by tinnitus but not vertigo. Some scholars believe that the main cause of acute deafness is middle ear disease. Both are currently treated with hormones. In the treatment of sudden deafness, hormones help to interrupt the pathological damage process and protect and promote the recovery of various inner ear functions. However, their effectiveness is limited for treating irreversible damage to inner ear tissues and organs. Hormones are particularly effective for patients with severe sudden deafness, especially those with mid- and low-frequency hearing loss. Internationally, glucocorticoids have been adopted as the standard treatment for sudden deafness.

[0003] Dexamethasone, as a synthetic adrenocortical hormone, has a strong anti-inflammatory effect on various organ systems; and compared with hydrocortisone and its derivatives, it has the weakest water and sodium retention side effect and the strongest anti-inflammatory effect. The biological half-life of dexamethasone is 36-54 hours, making it a long-acting glucocorticoid (Chinese Medical Abstracts Otorhinolaryngology, 2014, 29(5): 280). Currently, oral dexamethasone tablets or intratympanic injection of dexamethasone sodium phosphate are commonly used in clinical practice to treat acute deafness. However, oral administration has significant systemic side effects, and the drug is difficult to cross the blood-labyrinth barrier to reach the inner ear and exert its effect. Furthermore, excipients in the preparation are prone to remain in the inner ear and cannot be metabolized, thereby causing inflammation and other inner ear diseases. Many injection solutions are sterilized by high-pressure steam or flowing steam, which easily damages the stability of the drug and strengthens the interaction between raw materials and excipients, making related substances difficult to control. However, studies have shown that dexamethasone can enter the inner ear via the tympanic membrane-tympanic cavity route (Journal of Audiology and Speech Pathology, 2005, 13(4): 260-263), thus providing a new approach to the treatment of inner ear diseases. Compared with intravenous infusion, intratympanic injection can achieve an overall effective rate of 92.5% in the treatment of sudden deafness, which is significantly better than intravenous administration (Jilin Medical Journal, 2013, 34(27): 5607).

[0004] Currently available commercially available dexamethasone sodium phosphate injections have low concentrations, making them unsuitable for the high concentrations required for tympanic cavity administration. Furthermore, simply increasing the concentration of the lyophilized formulation cannot solve the dosage problem. Existing commercially available dexamethasone sodium phosphate lyophilized formulations all contain excipients such as mannitol, lactose, and sorbitol. In addition, to increase product stability, antioxidants and other substances are often added. These substances can easily remain in the ear, potentially causing inner ear inflammation, making them unsuitable for tympanic cavity administration. Summary of the Invention

[0005] In some embodiments, the present invention provides a lyophilized powder injection formulation, the formulation being a dexamethasone-based lyophilized powder injection, which does not contain a lyophilization support, excipient, or scaffold agent, and the lyophilized powder injection formulation contains a pharmaceutically acceptable excipient, glycerol, which is added before the lyophilization of the lyophilized powder injection formulation.

[0006] In some implementations, the excipients also include a pH adjuster.

[0007] In some implementations, the excipients do not include chelating agents or antioxidants.

[0008] In some embodiments, the dexamethasone drug is selected from one or more of dexamethasone, pharmaceutically acceptable derivatives of dexamethasone, or their salts or esters.

[0009] In some embodiments, the dexamethasone-based drugs include one or more of dexamethasone, dexamethasone sodium phosphate, dexamethasone acetate, or their salts.

[0010] In some embodiments, the dexamethasone sodium phosphate has a mass percentage of 60.0% to 99.9%.

[0011] In some embodiments, the dexamethasone sodium phosphate has a mass percentage of 60.0% to 95%.

[0012] In some embodiments, the dexamethasone sodium phosphate has a mass percentage of 60.0% to 90%.

[0013] In some embodiments, the dexamethasone sodium phosphate has a mass percentage of 70.0% to 90%.

[0014] In some embodiments, the mass ratio of dexamethasone to glycerol in the lyophilized powder injection is 8:1 to 2:1.

[0015] In some embodiments, the mass ratio of dexamethasone to glycerol in the lyophilized powder injection is 8:1 to 2.5:1.

[0016] In some embodiments, the mass ratio of dexamethasone to glycerol in the lyophilized powder injection is 6:1 to 6:2.4.

[0017] In some embodiments, the mass ratio of dexamethasone to glycerol in the lyophilized powder injection is 6:1.4 to 6:2.

[0018] In some embodiments, the formulation is prepared by freeze-drying a combination solution of dexamethasone, water, and pharmaceutically acceptable excipients.

[0019] In some embodiments, the dexamethasone drug is selected from one or more of dexamethasone, pharmaceutically acceptable derivatives of dexamethasone, or their salts or esters.

[0020] In some implementations, the excipients also include a pH adjuster.

[0021] In some embodiments, the osmotic pressure of the combined solution is 60 mOsm / kg to 449 mOsm / kg.

[0022] In some embodiments, the osmotic pressure of the combined solution is 150 mOsm / kg to 300 mOsm / kg.

[0023] In some embodiments, the osmotic pressure of the combined solution is 240 mOsm / kg to 300 mOsm / kg.

[0024] In some embodiments, the dexamethasone-based drugs include one or more of dexamethasone, dexamethasone sodium phosphate, dexamethasone acetate, or their salts.

[0025] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 15–80 mg / mL.

[0026] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 25–75 mg / mL.

[0027] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 25–65 mg / mL.

[0028] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 20–55 mg / mL.

[0029] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 20–45 mg / mL.

[0030] In some embodiments, the content of dexamethasone sodium phosphate in the combined solution is 30 mg / mL.

[0031] In some embodiments, the glycerol content is 0.01 to 3% w / v.

[0032] In some embodiments, the glycerol content is 0.01 to 2% w / v.

[0033] In some embodiments, the glycerol content is 0.01 to 1.45% w / v.

[0034] In some embodiments, the glycerol content is 0.01 to 1.2% w / v.

[0035] In some embodiments, the pH adjuster is selected from one or more of sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium hydroxide, hydrochloric acid, citric acid, or potassium dihydrogen phosphate.

[0036] In some embodiments, the pH adjuster added to the combined solution is 0.05–0.2 mm hydrochloric acid.

[0037] In some embodiments, the lyophilized powder injection is used after being reconstituted with a solvent, which includes water, an aqueous glycerol solution, or an aqueous sodium hyaluronate solution.

[0038] In some embodiments, the pH range of the combined solution is 6 to 9.

[0039] In some embodiments, the pH range of the combined solution is 7 to 9.

[0040] In some embodiments, the pH range of the combined solution is 7 to 8.5.

[0041] In some embodiments, the pH range of the combined solution is 7.5 to 8.2.

[0042] In some implementations, the pH range of the reconstituted drug solution is 6 to 9.

[0043] In some implementations, the pH range of the reconstituted drug solution is 7 to 9.

[0044] In some implementations, the pH range of the reconstituted drug solution is 7 to 8.5.

[0045] In some embodiments, the pH value of the reconstituted drug solution is in the range of 7.5 to 8.2. In some embodiments, the osmotic pressure of the combined solution before lyophilization is 60 mOsm / kg to 449 mOsm / kg.

[0046] In some embodiments, the osmotic pressure of the combined solution before lyophilization of the formulation is 150 mOsm / kg to 300 mOsm / kg.

[0047] In some embodiments, the osmotic pressure of the combined solution before lyophilization of the formulation is 240 mOsm / kg to 300 mOsm / kg.

[0048] In some embodiments, the formulation is an ear injection.

[0049] In some implementations, the formulation is an inner ear injection or a tympanic cavity injection. Domestic databases show 89 records of substandard quality for commercially available dexamethasone sodium phosphate injections in recent years, mainly due to visible foreign matter and related substances. For inner ear injections, the low levels of body fluids and enzymes in the inner ear make metabolism difficult, allowing impurities and excipients to remain, potentially affecting hearing. In the absence of other antioxidants or excipients, the injection solution is susceptible to oxidation or hydrolysis, also affecting the stability of the active ingredient.

[0050] As can be seen from the existing technology, obtaining a dexamethasone injection that meets the requirements (high efficacy, good stability, and few impurities or related substances) is not an easy task.

[0051] During the research on dexamethasone injection, the inventors discovered in their early studies that when dexamethasone is lyophilized, glycerol, a preservative commonly used for the freeze-drying of red blood cells, cannot be formed after being added to the lyophilization of the injection solution, thus affecting the lyophilization effect of dexamethasone.

[0052] In some implementation schemes, what is remarkable is that the inventors were not limited by previous research, but instead pursued excellence and obtained a drug that does not contain lyophilized support agents or excipients, and can improve drug stability by adding only a small amount of glycerol. At the same time, it can also adjust the osmotic pressure to make the drug more closely resemble the physiological osmotic pressure when administered into the tympanic cavity and prolong the duration of local drug action, thereby increasing the safety and effectiveness of tympanic cavity administration.

[0053] In some implementations, what is particularly remarkable is that the formulation obtained by the present invention can produce ideal white, loose freeze-dried blocks without the skeleton collapsing due to the addition of glycerol.

[0054] In some embodiments, the present invention provides a pharmaceutical kit comprising: (1) the lyophilized powder injection formulation; and (2) a solvent; wherein the lyophilized powder injection formulation and the solvent are packaged separately in the pharmaceutical kit.

[0055] In some implementations, the drug delivery device of the drug kit includes one or more of the following: needles and syringes, pumps, and microinjection devices.

[0056] In some implementations, the drug delivery method of the kit includes one or more of microsiphon / microchip delivery, microinfusion pump delivery, and reciprocating microfluidic delivery.

[0057] In some embodiments, the solvent includes water, an aqueous solution of glycerol, or an aqueous solution of sodium hyaluronate.

[0058] In some embodiments, the present invention provides a method for preparing the lyophilized powder injection formulation, wherein glycerol is added to water to obtain a glycerol aqueous solution; a dexamethasone-like drug is added to the glycerol aqueous solution, water for injection is added, and then freeze-drying is performed.

[0059] In some implementations, the freeze-drying process used in this invention can solve the problem of product shrinkage caused by the addition of glycerin.

[0060] In some embodiments, the freeze-drying steps include: pre-freezing, followed by a first drying, and then desorption drying; the first drying is performed at a temperature of -50°C to 10°C for a time of 1000 min to 2200 min.

[0061] In some embodiments, the temperature of the primary drying is -30°C to 0°C, and the time is 1000 min to 2000 min.

[0062] In some embodiments, the temperature of the primary drying is -30°C to 0°C, and the time is 1000 min to 1800 min.

[0063] In some embodiments, the temperature of the primary drying is -20°C to 0°C, and the time is 1000 min to 1500 min.

[0064] In some embodiments, the primary drying step includes: a temperature of -30°C to -15°C for 500 min to 2000 min; a temperature increase at a rate of 0.1 to 0.2°C / min to -14°C to -6°C for 300 to 500 min; and a temperature increase at a rate of 0.1 to 0.2°C / min to -5°C to 10°C for 100 min to 200 min.

[0065] In some embodiments, the primary drying step includes: a temperature of -30°C to -18°C for 500 min to 1200 min; a temperature increase at a rate of 0.1 to 0.2°C / min to -14°C to -8°C for 300 to 400 min; and a temperature increase at a rate of 0.1 to 0.2°C / min to -5°C to 5°C for 100 min to 150 min.

[0066] In some implementations, the pre-freezing temperature is -65°C to -20°C, and the time is 50 min to 150 min.

[0067] In some implementations, the pre-freezing temperature is -65°C to -25°C, and the time is 60 min to 500 min.

[0068] In some implementations, the pre-freezing temperature is -65°C to -25°C, and the time is 670 min to 400 min.

[0069] In some implementations, the pre-freezing temperature is -50°C to -35°C, and the time is 60-70 min to 200 min.

[0070] In some implementations, the pre-freezing temperature is -65°C to -25°C, and the time is 60 min to 120 min.

[0071] In some implementations, the pre-freezing temperature is -50°C to -35°C, and the time is 60 min to 120 min.

[0072] In some implementations, the analytical drying temperature is 25°C to 45°C, and the time is 600 min to 700 min.

[0073] In some implementations, the analytical drying temperature is 25°C to 40°C, and the time is 600 min to 700 min.

[0074] In some embodiments, the analytical drying step includes: heating to 25°C to 50°C at a rate of 1 to 6°C / min for a duration of 600 to 800 min.

[0075] In some embodiments, the analytical drying step includes: heating to 25°C to 40°C at a rate of 1 to 3°C / min for a duration of 600 to 700 min.

[0076] In some implementations, the freeze-drying step further includes: evacuating the vacuum after desorption drying.

[0077] In some implementation schemes, a vacuum of 0.1 to 0.2 mbar is used.

[0078] In some embodiments, the freeze-drying time is 1500 min to 5000 min.

[0079] In some embodiments, the freeze-drying time is 1500 min to 4000 min.

[0080] In some embodiments, the freeze-drying time is 1500 min to 3000 min.

[0081] In some embodiments, the present invention provides a lyophilized powder injection formulation obtained by the method described above.

[0082] In some embodiments, the present invention provides the use of the described lyophilized powder injection formulation, the described pharmaceutical kit, or the described preparation method in the preparation of medicaments for the prevention or treatment of ear diseases. Detailed Implementation

[0083] The following specific embodiments further illustrate the technical solution of the present invention. These specific embodiments do not represent a limitation on the scope of protection of the present invention. Non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the scope of protection of the present invention.

[0084] Some definitions

[0085] As used herein, improvement or relief of symptoms of a particular ear disease, condition, or symptom by administration of a particular formulation or medicine kit means any reduction in severity, delay in onset, slowing of progression, or shortening of duration attributable to or caused by the administration of the formulation or medicine kit, whether such effects are permanent or temporary, lasting or transient.

[0086] As used herein and in the appended claims, the singular forms “a / an,” “a / an,” and “the” include plural references unless the context clearly specifies otherwise. Thus, for example, reference to “a method” includes multiple such methods, and reference to “the segment” includes reference to one or more segments and their equivalents known to those skilled in the art, and so on.

[0087] Furthermore, unless otherwise stated, the use of "or" means "and / or". Similarly, "contains" and "includes" are interchangeable and are not intended to be restrictive.

[0088] It should be further understood that, when the term “comprising” is used to describe various embodiments, those skilled in the art will understand that, in certain specific circumstances, the language “substantially consisting of” or “consisting of” may be used instead to describe the embodiments.

[0089] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. While many methods and reagents are similar to or equivalent to those described herein, exemplary methods and materials are disclosed herein.

[0090] It should be understood that this disclosure is not limited to the specific methods, schemes, and reagents described herein, and these methods are subject to change. The terminology used herein is for the purpose of describing specific embodiments or aspects only and is not intended to limit the scope of this disclosure.

[0091] "Antioxidant" is an otomedically acceptable antioxidant and includes, for example, butylated hydroxytoluene (BHT), sodium ascorbate, ascorbic acid, sodium metabisulfite, and tocopherol. In some embodiments, antioxidants enhance chemical stability when needed. Antioxidants are also used to counteract the ototoxic effects of certain therapeutic agents, including those used in combination with the corticosteroids disclosed herein.

[0092] The "inner ear" refers to the inner ear, which includes the cochlea, vestibular labyrinth, and the round window connecting the cochlea to the middle ear.

[0093] The "round window membrane" is the membrane that covers the cochlear window (also known as the circular window, perfect round window, or round window) in humans. In humans, the round window membrane is about 70 micrometers thick.

[0094] "Pharmacodynamics" refers to the factors that determine the biological response observed at the target site within the target ear structure relative to the drug concentration.

[0095] "Pharmacokinetics" refers to the factors that determine the achievement and maintenance of appropriate drug concentrations at the desired sites within the target ear structure.

[0096] As used herein, the term “treatment” includes preventive and / or therapeutic relief, mitigation or improvement of symptoms of a disease or condition, prevention of other symptoms, improvement or prevention of underlying metabolic causes of symptoms, suppression of a disease or condition (e.g., prevention of disease or condition progression), reduction of a disease or condition, resolution of a disease or condition, relief of symptoms caused by a disease or condition, or termination of symptoms of a disease or condition.

[0097] In some embodiments, the present invention provides a pharmaceutical kit for treating tumors, comprising the aforementioned lyophilized powder for injection and a solvent. In the pharmaceutical kit, the lyophilized powder for injection is not necessarily, and usually is not mixed with the solvent, but is typically packaged separately. The separately packaged lyophilized powder for injection and the solvent may also contain their respective adjuvants. The adjuvant refers to a means in pharmacology that can assist in the efficacy of a drug. The pharmaceutical kit may also comprise individually packaged lyophilized powder for injection and individually packaged solvent.

[0098] As used herein, the term “pharmaceutically acceptable” means compounds, materials, compositions, and / or dosage forms that (within the scope of reasonable medical judgment) are suitable for contact with a patient’s tissues without causing additional toxicity, irritation, allergic reactions, or other problems or complications (i.e., with a reasonable risk / benefit ratio).

[0099] The term “prevention” has a commonly accepted meaning in the field, and when used in connection with a condition, disease, syndrome or any other medical condition, it has a meaning well-known in the field, including the administration of a drug in which the frequency of the occurrence of the medical condition is reduced or the onset or symptoms are delayed in patients who are given the drug compared to patients who are not given the drug.

[0100] The term “treatment” as used in this article includes reversing, alleviating or suppressing the symptoms, clinical features and underlying pathology of a disease in a way that improves or stabilizes the patient’s condition.

[0101] The term "prevention or treatment" has the generally accepted meaning in the art, including administering one or more of the present invention's drugs to a recipient. If the treatment is administered before the clinical manifestation of an adverse condition (e.g., to an animal with a disease or other adverse condition), then the treatment is preventative (i.e., to prevent the recipient from developing an adverse condition); conversely, if the treatment is administered after the clinical manifestation of an adverse condition, then the treatment is therapeutic (i.e., to reduce, improve, or stabilize an existing adverse condition or its side effects).

[0102] In the embodiments described herein, the freeze-drying technology is operated according to the freeze dryer operating procedures, and the freeze-drying curve 1 (specific details are based on curve 1 in Table 13 of Experimental Example 3) is set with reference to Table 1 below.

[0103] Table 1

[0104]

[0105]

[0106] Example 1: Formulation and preparation method of dexamethasone sodium phosphate injection

[0107] Table 2

[0108]

[0109] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0110] Example 2: Formulation and preparation method of dexamethasone sodium phosphate injection

[0111] Table 3

[0112]

[0113] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0114] Example 3: Formulation and preparation method of dexamethasone sodium phosphate injection

[0115] Table 4

[0116]

[0117] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0118] Example 4: Formulation and preparation method of dexamethasone sodium phosphate injection

[0119] Table 5

[0120]

[0121] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0122] Example 5: Formulation and preparation method of dexamethasone sodium phosphate injection

[0123] Table 6

[0124]

[0125]

[0126] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0127] Example 6: Formulation and preparation method of dexamethasone sodium phosphate injection

[0128] Table 7

[0129]

[0130] Preparation method: Add glycerin to 70% of the prescribed amount of water according to the above prescription table to obtain a glycerin-water solution; add dexamethasone sodium phosphate to the glycerin-water solution, adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection. After the drug is completely dissolved, sterilely filter and dispense 1ml into each vial, half-stopping. Place in a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and seal with an aluminum cap to obtain the sterile lyophilized preparation.

[0131] Example 7: Formulation and preparation method of dexamethasone sodium phosphate for injection (containing chelating agent EDTA-2NaCa calcium sodium edetate).

[0132] Table 8

[0133]

[0134] Preparation method: Dissolve the prescribed amount of dexamethasone sodium phosphate and excipient EDTA-2NaCa in room temperature water for injection. Adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection as needed. After the drug is completely dissolved, sterilely filter the solution and dispense 1ml into vials, partially stoppering each vial. Place the vials into a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and cap with aluminum to obtain the sterile lyophilized formulation.

[0135] Example 8: Formulation and preparation method of dexamethasone sodium phosphate for injection (containing the antioxidant sodium bisulfite)

[0136] Prescription composition:

[0137] Table 9

[0138]

[0139] Preparation method: Dissolve the prescribed amount of dexamethasone sodium phosphate and excipient sodium bisulfite in 70% water for injection. Adjust the pH of the solution to 8.0 with 0.1M NaOH, and add water for injection as needed. After the drug is completely dissolved, sterilely filter the solution and dispense 1ml into vials, partially stoppering each vial. Place the vials into a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and cap with aluminum to obtain the sterile lyophilized formulation.

[0140] Example 9: Formulation and preparation method of dexamethasone sodium phosphate for injection (containing hyaluronic acid)

[0141] Table 10

[0142]

[0143] Preparation method: Dissolve the prescribed amount of dexamethasone sodium phosphate and excipient hyaluronic acid in 70% water for injection. Adjust the pH of the solution to 8.0 with 0.1M HCl, and add water for injection as needed. After the drug is completely dissolved, sterilely filter the solution and dispense 1ml into vials, partially stoppering each. Place the vials into a lyophilizer for freeze-drying using lyophilization curve 1. After freeze-drying, purge with nitrogen, stopper, and cap with aluminum to obtain the sterile lyophilized formulation.

[0144] Experimental Example 1: Examination of the appearance and osmotic pressure of the freeze-dried product

[0145] The dexamethasone sodium phosphate formulations (lyophilized powder for injection) prepared according to Examples 1 to 9 were tested for moisture content, osmotic pressure, and appearance. The results are shown in Table 11.

[0146] Table 11

[0147]

[0148] As can be seen from the appearance and osmotic pressure results in the table above, glycerol was added in Examples 1 to 6. The greater the amount of glycerol added, the greater the impact on the formulation. When the ratio of dexamethasone sodium phosphate to glycerol was 6:3, the lyophilized block collapsed.

[0149] In Example 9, the addition of hyaluronic acid severely affected the formulation. Furthermore, glycerin is an osmotic pressure regulator; the formulation without glycerin had a significantly lower osmotic pressure, only around 150 mOsm / kg. When the ratio of dexamethasone sodium phosphate to glycerin reached 6:1 to 6:3, the osmotic pressure was 240 to 300 mOsm / kg, close to physiological osmotic pressure.

[0150] Example 2: Investigation of related substances in high-temperature testing of freeze-dried products

[0151] The dexamethasone sodium phosphate formulations prepared according to Examples 1 to 8 were removed from their outer packaging and placed in a high-temperature (60°C) stability test chamber. Samples were taken on days 0, 5, and 10 to examine the content of related substances (including: degradation impurity dexamethasone and total impurities). The results are shown in Table 12.

[0152] Table 12

[0153]

[0154]

[0155] The results of the total impurities and content in the table above show that the product prepared with the addition of glycerin has significantly improved stability compared to the product without glycerin. The total impurity content is significantly lower than that of the formulations with chelating agent EDTA and antioxidants, while the dexamethasone sodium phosphate content is significantly higher than that of the formulations with chelating agent EDTA and antioxidants.

[0156] Comparison of different process examples in Experiment 3 (Examples 4, 10, 11, and 12)

[0157] The formulation used in Experimental Example 3 is shown in Table 13 below (Formulation of Example 4).

[0158] Table 13

[0159]

[0160] Preparation method: Glycerin was added to 70% of the prescribed amount of water to obtain a glycerin-water solution; dexamethasone sodium phosphate was added to the glycerin-water solution, the pH of the solution was adjusted to 8.0 with 0.1M HCl, and water for injection was added. After the drug was completely dissolved, it was sterilely filtered and dispensed into vials at 1ml per vial, half-stopped. The vials were then sent to a lyophilizer for freeze drying, using the freeze-drying curves in Table 14 below. After freeze-drying, nitrogen gas was applied, the vials were stopped, and aluminum caps were formed to obtain the sterile freeze-dried preparation.

[0161] Table 14

[0162]

[0163]

[0164] The results of the appearance and moisture content analysis of the freeze-dried product are shown in Table 15.

[0165] Table 15

[0166]

[0167] The results in the table above show that the primary drying temperature and drying time in the lyophilization curve parameters have a significant impact on the 1.0% (w / v) glycerol formulation. The primary drying temperature should be as low as possible, and the drying time should be as long as possible, with a total lyophilization time of over 2000 min.

[0168] Experimental Example 4: Pharmacodynamic Study of Dexamethasone Sodium Phosphate for Injection (Glycerin Formulation)

[0169] Take the dexamethasone sodium phosphate formulations prepared according to Examples 1 to 6 (containing glycerol in the formulation) and the dexamethasone sodium phosphate formulation prepared in Example 7 (containing the chelating agent EDTA-2NaCa calcium sodium edetate). Before the experiment, add water for injection to each of the above formulations to prepare a 30 mg / ml dexamethasone sodium phosphate test solution for later use. Separately, take 5 mg of commercially available dexamethasone sodium phosphate for injection and add water for injection to prepare a 5 mg / ml dexamethasone sodium phosphate test solution for later use.

[0170] The efficacy of the above-mentioned dexamethasone sodium phosphate experimental group against sudden deafness was evaluated using a noise-induced deafness model commonly used in new drug evaluation. A model group and a blank control group (water for injection) were also set up.

[0171] 1. Guinea pig noise-induced deafness model:

[0172] Normal-hearing guinea pigs were divided into 9 groups: The groups were subjected to white noise stimulation at 120 dB for 1 hour. On the day of stimulation, the animals in the drug treatment group were anesthetized with ketamine and toluidine via intraperitoneal injection according to their body weight. Seven of these groups received a single intratympanic injection of different drugs from Examples 1 to 7 in each ear, while one group received a single intratympanic injection of a commercially available drug in each ear. The model group received white noise stimulation at 120 dB for 1 hour without drug treatment. Fifteen days after stimulation, the guinea pigs' hearing had not recovered to normal levels, exhibiting significant hearing loss, indicating successful noise modeling.

[0173] Hearing thresholds (dB) were assessed before and 15 days after noise exposure using auditory brainstem response (ABR) testing. The results are shown in Table 16. Before noise exposure, the average hearing of all groups was within the normal range of 21-26.1 dB. Immediately after noise exposure, the average hearing in the model group was 94-95 dB (total deafness), while the average hearing in the drug-treated group and the commercially available product group was between 29-50 dB after 15 days of administration, indicating that dexamethasone sodium phosphate treatment had a certain protective effect on hearing. The 30 mg / mL dexamethasone sodium phosphate group showed better efficacy than the commercially available 5 mg / mL dexamethasone sodium phosphate group.

[0174] Table 16 Hearing Threshold Assessment Table for Guinea Pig Noise-Induced Deafness Model Experiment (dB: Mean ± Standard Deviation)

[0175] Before the noise Noise Immediately Administer for 3 days Administer for 5 days Administer for 7 days Administer for 10 days Administer for 15 days Model group 22.5±0.0 94.7±0.0 87.3±15.1 85.6±9.5 84.8±18.2 82.8±22.3 79.8±23.9 Example 1 24.4±0.0 94.2±19.5 65.4±18.0 59.1±25.8 51.8±22.1 48.2±19.0 36.8±12.8 Example 2 23.2±0.0 94.8±16.8 68.1±21.4 55.0±23.0 53.0±24.5 44.3±20.5 33.6±24.7 Example 3 22.8±0.0 94.7±9.5 67.3±22.0 56.8±23.7 49.3±24.6 45.8±20.8 31.7±24.5 Example 4 26.1±0.0 94.4±18.0 60.5±22.0 53.0±22.3 50.5±22.0 46.4±24.5 29.7±23.0 Example 5 24.6±0.0 93.8±10.2 59.1±25.8 55.8±28.7 53.4±23.3 43.6±19.9 37.7±9.5 Example 6 23.2±0.0 94.3±15.1 65.4±18.0 55.0±23.0 51.8±23.9 47.5±28.0 35.6±18.9 Example 7 25.6±0.0 95.0±18.0 61.1±21.4 60.8±18.8 54.3±20.9 51.8±22.1 49.1±25.8 Commercial Group 21.8±0.0 94.8±24.0 74.2±19.5 67.2±15.9 54.5±28.0 47.7±23.0 40.8±21.4

[0176] Experimental Example 5: Inner Ear Residue Test

[0177] Take the dexamethasone sodium phosphate preparations prepared according to Examples 1 to 9 above, and add water for injection to prepare a 30 mg / ml dexamethasone sodium phosphate test solution (Examples 1-9 in Table 17) for later use. Separately, take commercially available dexamethasone sodium phosphate for injection (5 mg), and add water for injection to prepare a 5 mg / ml dexamethasone sodium phosphate test solution for later use (commercially available group in Table 17). Simultaneously prepare 2% w / v glycerol reconstituted solution and 25% w / v glycerol reconstituted solution.

[0178] Preparation method of 2% w / v glycerol aqueous reconstituted solution (2% w / v glycerol aqueous group): Take 2g of glycerol and add water for injection to 100ml to prepare 2% w / v glycerol aqueous reconstituted solution.

[0179] Preparation method of 25% w / v glycerol aqueous reconstituted solution (25% w / v glycerol water): Take 25g of glycerol and add water for injection to 100ml to prepare a 2% w / v glycerol aqueous reconstituted solution.

[0180] Inner ear residue test: After guinea pigs were modeled according to the method in Example 4, they were anesthetized and then injected intratympanically with different formulations of dexamethasone sodium phosphate, the commercially available test solution, and 2% w / v glycerol reconstituted solution and 25% w / v glycerol reconstituted solution. Residue and inflammation in the middle ear were observed 3 and 10 days after administration. The results are shown in Table 17. Formulations with a glycerol content of 2 w / v% or less did not produce inner ear residue or inflammation. When the glycerol content reached 25%, significant residue and inflammation appeared in the guinea pig inner ear, and hearing was impaired. Furthermore, formulations containing hyaluronic acid and commercially available products containing mannitol also produced residue in the inner ear initially, and formulations containing sodium bisulfite may induce inflammation in the guinea pig inner ear.

[0181] Table 17 Middle ear residual and inflammation after a single intratympanic injection of various dexamethasone sodium phosphates.

[0182]

[0183]

Claims

1. A dexamethasone sodium phosphate lyophilized powder for injection, characterized in that, The dexamethasone sodium phosphate lyophilized powder for injection is composed of dexamethasone sodium phosphate, water for injection, glycerol, and a pH adjuster; the dexamethasone sodium phosphate lyophilized powder for injection is prepared by lyophilizing a combined solution of dexamethasone sodium phosphate, water for injection, glycerol, and a pH adjuster; the pH of the combined solution is 6-8.5; the glycerol content in the combined solution is 0.5-1.0% w / v; the mass ratio of dexamethasone sodium phosphate to glycerol in the dexamethasone sodium phosphate lyophilized powder for injection is 6:1 to 6:2; the glycerol is added before lyophilizing the dexamethasone sodium phosphate lyophilized powder for injection; the dexamethasone sodium phosphate lyophilized powder for injection does not contain lyophilization support, excipients, scaffolding agents, chelating agents, or antioxidants.

2. The dexamethasone sodium phosphate lyophilized powder for injection as described in claim 1, characterized in that, The mass ratio of dexamethasone sodium phosphate to glycerin in the lyophilized dexamethasone sodium phosphate injection formulation is 6:1.4 to 6:

2.

3. The dexamethasone sodium phosphate lyophilized powder for injection as described in claim 1, characterized in that, The pH adjuster is selected from one or more of sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium hydroxide, hydrochloric acid, citric acid, or potassium dihydrogen phosphate.

4. The dexamethasone sodium phosphate lyophilized powder for injection as described in claim 1, characterized in that, The pH adjuster added to the combined solution is 0.05–0.2 M hydrochloric acid.

5. The lyophilized powder injection formulation as described in claim 1, characterized in that, The pH range of the combined solution is 6 to 8.

6. The lyophilized powder injection formulation as described in claim 1, characterized in that, The pH range of the combined solution is 7 to 8.

7. The dexamethasone sodium phosphate lyophilized powder for injection as described in claim 1, characterized in that, The dexamethasone sodium phosphate lyophilized powder for injection is used after being reconstituted with a solvent, namely water.

8. The dexamethasone sodium phosphate lyophilized powder for injection as described in claim 1, characterized in that, The dexamethasone sodium phosphate lyophilized powder for injection is a tympanic cavity injection.

9. A medicine kit, characterized in that, include: (1) The dexamethasone sodium phosphate lyophilized powder for injection according to any one of claims 1 to 8; (2) Solvent; In the aforementioned pharmaceutical kit, the dexamethasone sodium phosphate lyophilized powder for injection and the solvent are packaged separately.

10. The pharmaceutical kit as described in claim 9, characterized in that, The drug delivery device of the drug kit includes one or more of the following: needle and syringe, pump, and microinjection device.

11. The pharmaceutical kit as described in claim 9, characterized in that, The drug delivery system includes one or more of the following methods: microsiphon / microchip delivery, microinfusion pump injection, and reciprocating microfluidic delivery.

12. The pharmaceutical kit as described in claim 9, characterized in that, The solvent is water.

13. A method for preparing the dexamethasone sodium phosphate lyophilized powder for injection according to any one of claims 1-8, characterized in that the steps include... include: Glycerin was added to water to obtain a glycerin aqueous solution; dexamethasone sodium phosphate was added to the glycerin aqueous solution, water was added, and then freeze-drying was performed. The freeze-drying steps include: pre-freezing, followed by a first drying, and then desorption drying; the first drying temperature is -50℃ to 10℃, and the time is 1000 min to 2200 min.

14. The method as described in claim 13, characterized in that, The temperature for the first drying step is -30℃ to 0℃, and the time is 1000 min to 2000 min.

15. The method as described in claim 13, characterized in that, The temperature for the first drying step is -30℃ to 0℃, and the time is 1000 min to 1800 min.

16. The method as described in claim 13, characterized in that, The temperature for the first drying step is -20℃ to 0℃, and the time is 1000 min to 1500 min.

17. The method as described in claim 13, characterized in that, The drying process includes: a temperature of -30℃ to -15℃ for 500 min to 2000 min; a temperature increase of 0.1 to 0.2℃ / min to -14℃ to -6℃ for 300 to 500 min; and a temperature increase of 0.1 to 0.2℃ / min to -5 to 10℃ for 100 min to 200 min.

18. The method as described in claim 13, characterized in that, The drying process includes: a temperature of -30℃ to -18℃ for 500 min to 1200 min; a temperature increase of 0.1 to 0.2℃ / min to -14℃ to -8℃ for 300 to 400 min; and a temperature increase of 0.1 to 0.2℃ / min to -5℃ to 5℃ for 100 min to 150 min.

19. The method as described in claim 13, characterized in that, The pre-freezing temperature is -65℃ to -20℃, and the time is 50 min to 150 min.

20. The method as described in claim 13, characterized in that, The pre-freezing temperature is -65℃ to -25℃, and the time is 60 min to 500 min.

21. The method as described in claim 13, characterized in that, The pre-freezing temperature is -65℃ to -25℃, and the time is 70 min to 400 min.

22. The method as described in claim 13, characterized in that, The pre-freezing temperature is -50℃ to -35℃, and the time is 70 min to 200 min.

23. The method as described in claim 13, characterized in that, The analytical drying temperature is 25℃~45℃, and the time is 600 min~700 min.

24. The method as described in claim 13, characterized in that, The analytical drying temperature is 25℃~40℃, and the time is 600 min~700 min.

25. The method as described in claim 13, characterized in that, The analytical drying step includes: heating to 25°C~50°C at a rate of 1~6°C / min for a duration of 600 min~800 min.

26. The method as described in claim 13, characterized in that, The analytical drying step includes: heating to 25°C~40°C at a rate of 1~3°C / min for a duration of 600 min~700 min.

27. The method as described in claim 13, characterized in that, The freeze-drying step further includes: vacuuming after desorption drying.

28. The method as described in claim 13, characterized in that, Vacuum was drawn at a pressure of 0.1 mbar to 0.2 mbar.

29. The method as described in claim 13, characterized in that, The freeze-drying time is 1500 min to 5000 min.

30. The method as described in claim 13, characterized in that, The freeze-drying time is 1500 min to 4000 min.

31. The method as described in claim 13, characterized in that, The freeze-drying time is 1500 min to 3000 min.

32. The use of the dexamethasone sodium phosphate lyophilized powder injection formulation according to any one of claims 1-8, the pharmaceutical kit according to any one of claims 9-12, or the method according to any one of claims 13-31 in the preparation of a medicament for the prevention or treatment of sudden deafness.

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