Methods and compositions for treating hearing disorders

Abstract

The present invention provides methods and compositions for treating neurological disorders, such as hearing disorders. The method comprises intranasally administering a therapeutically effective amount of NMDP or a salt thereof to treat tinnitus or meniere's disease, or a symptom of tinnitus or meniere's disease, in a subject in need thereof. Compositions comprise an intranasal delivery formulation of a therapeutically effective amount of NMDP or a salt thereof for use in the treatment of neurological disorders.

Description

[0001] Cross-references

[0002] This application claims priority to U.S. Provisional Application No. 63 / 478,883, filed on January 6, 2023, the entire contents of which are incorporated herein by reference in their entirety. Background Art

[0003] Hearing impairment is a growing health problem with diverse and complex causes. While some types of hearing impairment have clear genetic causes, others are entirely or at least partially due to environmental factors.

[0004] Tinnitus is a common hearing impairment characterized by ringing in the ears. It can cause a range of symptoms, including fatigue, stress, sleep disturbances, poor concentration, memory problems, depression, anxiety, irritability, and headaches, making it a disabling condition. It affects approximately 20% of the population and is particularly common among the elderly or those exposed to high-decibel noise through occupational exposure.

[0005] Another hearing impairment is Meniere's disease, an inner ear disease that can cause tinnitus-like symptoms (such as ringing in the ears), hearing loss or loss, vertigo, and a feeling of fullness or blockage in the ears, but its cause is not necessarily related to noise exposure and may be genetic. Vertigo attacks can occur suddenly or after a brief period of tinnitus-like symptoms or blurred hearing. Some patients have long intervals between vertigo attacks, while others may have frequent attacks over several days. Some patients with Meniere's disease have vertigo symptoms that are so severe that they can even cause imbalance and falls. For patients, Meniere's disease is often a serious and disabling disease.

[0006] Currently, treatment options for hearing impairments such as tinnitus and Meniere's disease are extremely limited, and most existing therapies are non-drug treatments. These therapies do not directly address the persistent ringing in the ears, but instead focus on helping patients manage their condition subjectively, such as using white noise machines or psychological counseling. Therefore, there remains an urgent need to develop pharmaceutical compositions that can alleviate, improve, and / or combat one or more symptoms of tinnitus or Meniere's disease. Summary of the Invention

[0007] Disclosed herein are pharmaceutical compositions for treating hearing impairment. The present disclosure relates to a novel pharmaceutical formulation that delivers an effective dose of nimodipine (NMDP) via mucosal administration, such as intranasal administration, for treating hearing impairment.

[0008] It should be understood that mucosal administration has many advantages over conventional drug administration methods such as intravenous administration, parenteral injection or oral tablets. For example, patients do not need to arrange and remember the time to take the medicine, nor do they need to endure painful injections. The administration process is not affected by stomach or digestive problems. Mucosal administration can prevent the drug from being degraded in the gastrointestinal tract or liver. In addition, mucosal administration can quickly deliver the active agent to the subject, thereby quickly alleviating hearing impairment that requires emergency treatment in some cases (such as when the subject is driving a vehicle or doing work that requires auditory perception). However, due to the low water solubility of non-polar molecules (such as NMDP) and the small volume of the nasal cavity, the amount of solution used to deliver the raw material to the subject during intranasal administration is limited, so it is extremely difficult to formulate it into an intranasal preparation. This area still needs an NMDP intranasal preparation that is effective for treating hearing impairment and can quickly relieve the symptoms of hearing impairment.

[0009] One or more aspects disclosed herein include a pharmaceutical composition comprising a therapeutically effective amount of NMDP, the composition being formulated for intranasal administration, the composition comprising: a therapeutically effective amount of NMDP; and a surfactant, a solvent, and a water-soluble cellulose polymer. One or more aspects disclosed herein include a pharmaceutical composition comprising a therapeutically effective amount of NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a therapeutically effective amount of NMDP; and a liquid carrier, wherein the liquid carrier further comprises a surfactant, a water-soluble cellulose polymer, a solvent, and water. One or more aspects disclosed herein include a pharmaceutical composition comprising a therapeutically effective amount of NMDP, the composition being formulated for intranasal administration, the composition comprising: a therapeutically effective amount of NMDP, a surfactant, a water-soluble cellulose polymer, a solvent, and water. In some embodiments, the formulation further comprises ethanol. In some embodiments, the ethanol is present in an amount of up to 5% (w / w). In some embodiments, the surfactant is a polysorbate or a combination of polysorbates. In some embodiments, the surfactant is polysorbate 20 (TWEEN-20). In some embodiments, the surfactant is polysorbate 80 (TWEEN-80). In some embodiments, the content of the surfactant is at least 0.00001% and at most 2% (w / w). In some embodiments, the content of the surfactant is at least 0.05% (w / w) and at most 2% (w / w). In some embodiments, the water-soluble cellulose polymer may include hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC) and its sodium salt (CMCNa) or a combination thereof. In some embodiments, the content of the water-soluble cellulose polymer is at least 0.00001% and at most 2% (w / w). In some embodiments, the content of the water-soluble cellulose polymer is at least 0.05% (w / w) and at most 2% (w / w). In some embodiments, the solvent is PEG, an alkoxy derivative of polyethylene glycol, methoxypolyethylene glycol (mPEG), mPEG350, polyethylene glycol (PEG400) or a combination thereof. In some embodiments, the content of the solvent is at least 0.00001% and at most 94% (w / w). In some embodiments, the content of the solvent is at least 66% (w / w) and at most 94% (w / w). In some embodiments, the content of water is at least 0.0% and at most 50% (w / w). In some embodiments, the content of water is at least 0.0% and at most 20% (w / w). In some embodiments, the content of water is at least 0.00001% (w / w) and at most 50% (w / w). In some embodiments, the content of water is at least 0.0001% (w / w) and at most 20% (w / w).In some embodiments, the surfactant is a nonionic surfactant. In some embodiments, the nonionic surfactant is a polysorbate or a combination of polysorbates. In some embodiments, the surfactant is polysorbate 20 (TWEEN-20). In some embodiments, the surfactant is polysorbate 80 (TWEEN-80). In some embodiments, the content of polysorbate 80 (TWEEN-80) is at least 0.00001% and at most 2% (w / w). In some embodiments, the content of polysorbate 80 (TWEEN-80) is at least 0.05% (w / w) and at most 2% (w / w). In some other embodiments, the content of TWEEN-80 is less than about 2.5%. In some embodiments, the water-soluble cellulose polymer is HPMC, HEC, CMC, sodium carboxymethylcellulose or a combination thereof. In some embodiments, the water-soluble cellulose polymer only comprises HPMC. In some embodiments, the content of the HPMC is at least 0.00001% and at most 2% (w / w). In some embodiments, the content of the HPMC is at least 0.05% (w / w) and at most 2% (w / w). In some embodiments, the solvent is PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof. In some embodiments, the content of the PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof is at least 0.0% and at most 94% (w / w). In some embodiments, the content of the PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof is at least 66% (w / w) and at most 94% (w / w). In some embodiments, the content of the water is at least 0.00001% and at most 20% (w / w). In some embodiments, the water content is at least 0.00% (w / w) and at most 20% (w / w).

[0010] One or more aspects disclosed herein include a method for preparing a liquid NMDP composition, comprising the following steps: dissolving Tween 80 (<2% w / w) in mPEG350 and PEG400, respectively; mixing NMDP with the solution from step 1) to saturation solubility or a desired effective concentration; preparing an aqueous solution of HPMC or HPMC buffer (<2% w / w) with a pH range of about 6.4 to about 7.4; adding the HPMC aqueous solution or HPMC buffer from step 3) to the corresponding solution from step 2) with stirring, and allowing the mixture to stand to observe the stability of the solution or suspension. In some embodiments, the solution from step 2) is NMDP in PEG / Tween 80 solution or NMDP in mPEG / Tween 80 solution. In some embodiments, the concentration of NMDP is at least 68 mg / mL. In some embodiments, the concentration of NMDP is at least 0.1 mg / mL and up to 100 mg / mL. In some embodiments, the concentration of NMDP is 68 mg / mL. In some embodiments, the concentration of NMDP is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, or 45 mg / mL.

[0011] In some embodiments, the concentration of NMDP is 50 mg / mL. In some embodiments, the surfactant, water-soluble cellulose polymer, solvent, water, and ethanol constitute the liquid carrier. In some embodiments, the liquid carrier is less than 1000, 900, 800, 700, 600, 500, or 400 uL. In some embodiments, the liquid carrier is less than 300 uL. In some embodiments, the liquid carrier is less than 150 uL. In some embodiments, the NMDP is dissolved, suspended, or both dissolved and suspended in the liquid carrier.

[0012] Aspects disclosed herein provide a method for treating a neurological disease such as hearing disorder, comprising intranasally administering a therapeutically effective amount of NMDP or a salt thereof to a subject in need thereof, wherein the hearing disorder comprises tinnitus or Meniere's disease or a symptom of tinnitus or Meniere's disease.

[0013] Aspects disclosed herein provide a method for treating a neurological disorder, such as hearing impairment, comprising intranasally administering a therapeutically effective amount of NMDP or a salt thereof to a subject in need thereof, wherein the hearing impairment comprises tinnitus or Meniere's disease or a symptom of tinnitus or Meniere's disease. In some embodiments, the therapeutically effective amount of NMDP is from about 0.1 mg to about 5 mg per kilogram of the subject. In some embodiments, the method further comprises administering the pharmaceutical composition in a volume of from about 10 μl to about 300 μl per dose. In some embodiments, the method further comprises contacting at least a portion of the therapeutically effective amount of the pharmaceutical composition with the mucosa of at least one nasal cavity. In some embodiments, the method further comprises spraying a first amount of the pharmaceutical composition into a first nasal cavity, spraying a second amount of the pharmaceutical composition into a second nasal cavity, and optionally, after a preselected time delay, spraying a third amount of the pharmaceutical composition into the first nasal cavity. In some embodiments, the method further comprises administering at least a fourth amount of the pharmaceutical composition into the second nostril, optionally after a preselected time delay. In some embodiments, the treatment method achieves a bioavailability that is about 80% to 125% of the bioavailability achieved by intravenous administration of the same pharmaceutical composition. In some embodiments, the method further comprises administering the pharmaceutical composition at any time before or after the onset of tinnitus or Meniere's disease symptoms. In some embodiments, the method further comprises administering the pharmaceutical composition at least once a day, twice a day, three times a day, four times a day, five times a day, six times a day, seven times a day, eight times a day, nine times a day, or ten times a day. In some embodiments, a therapeutically effective amount of NMDP is administered intranasally every two hours. In some embodiments, a therapeutically effective amount of NMDP is administered intranasally every four hours. In some embodiments, the method further comprises administering the pharmaceutical composition weekly. In some embodiments, the method further comprises administering the pharmaceutical composition via low-dose therapy. In some embodiments, the method further comprises titrating the administration of the pharmaceutical composition based on vestibular symptoms. In some embodiments, the method further comprises administering the pharmaceutical composition for at least 1, 2, 3, 4, 5, 6, 14, 21, 28, 60, 120, or 400 days. In some embodiments, the method further comprises administering the pharmaceutical composition in unit dosage form. In some embodiments, the method further comprises administering the pharmaceutical composition so as not to cause changes in cochlear potentials. In some embodiments, the method further comprises increasing the subject's dose of the pharmaceutical composition until symptoms of the inner ear disorder are observed. In some embodiments, the symptoms of the inner ear disorder include spontaneous nystagmus observed with Frenzel glasses, balance disorders, motion intolerance, or hearing loss. In some embodiments, the method further comprises administering the pharmaceutical composition via a drug delivery device.In some embodiments, the effect of the pharmaceutical composition is measured by one of the following assessments: a) measuring a change in auditory brainstem response (ABR) threshold, b) a change in auditory speech recognition as measured by a speech-in-noise test, c) a change in auditory speech recognition as measured by a digit-in-noise test, d) a change in low-frequency hearing threshold, e) a change in the incidence of adverse events following administration of the pharmaceutical composition, f) a change in the severity of tinnitus or Meniere's disease, g) a change in the loudness of tinnitus or Meniere's disease, h) a change in the severity of vertigo, i) a change in the sensation of fullness in the ears, j) a change in dizziness, and k) a change in hair cell function observed by a change in ABR threshold following administration of the pharmaceutical composition. In some embodiments, the measurement of ABR threshold is performed in the frequency range of 200 Hz to 30 kHz.

[0014] Aspects disclosed herein provide a method of treating tinnitus or Meniere's disease, or a symptom of tinnitus or Meniere's disease, in a subject in need thereof, comprising: administering an L-type calcium channel blocker to the subject; assessing whether the subject responds to the L-type calcium channel blocker; and if the subject responds to the L-type calcium channel blocker, administering to the subject a therapeutically effective amount of NMDP or a salt thereof.

[0015] In some embodiments, the method further comprises administering a therapeutically effective amount of NMDP or a salt thereof to the patient intranasally to treat tinnitus or Meniere's disease or a symptom of tinnitus or Meniere's disease. In some embodiments, the therapeutically effective amount of NMDP is administered in a composition comprising: a therapeutically effective amount of NMDP; a carrier; a citrate buffer; and benzalkonium chloride. In some embodiments, the therapeutically effective amount of NMDP is administered in a composition comprising: a therapeutically effective amount of NMDP; about 30% to about 95% by weight of one or more natural or synthetic carriers or any combination thereof; about 10% to about 70% by weight of a non-aqueous solvent; and a surfactant. In some embodiments, the therapeutically effective amount of NMDP is administered in a composition comprising at least one of PEG, mPEG, or water. In some embodiments, the therapeutically effective amount of NMDP is administered in a composition comprising PEG, mPEG, and water. In some embodiments, a therapeutically effective amount of NMDP is administered in a composition comprising about 20% to about 90% by weight mPEG, about 10% to about 50% by weight PEG, about 5% to about 20% by weight water, and NMDP. In some embodiments, a therapeutically effective amount of NMDP is administered in a composition comprising about 70% by weight mPEG, about 20% by weight PEG, and about 10% by weight water.

[0016] Aspects disclosed herein provide a method for achieving a therapeutically effective area under the curve for NMDP extrapolated from administration time to infinity (AUC0-infinity) in a subject in need thereof, comprising intranasally administering a nasal pharmaceutical composition to the subject, wherein the nasal pharmaceutical composition comprises: about 0.1 mg to about 5 mg per kilogram of NMDP or a pharmaceutically acceptable salt thereof; a buffer; and a surfactant, wherein after administration of the nasal pharmaceutical composition to the subject, the subject exhibits an AUC0-infinity of NMDP between about 200 h*ng / mL and 400 h*ng / mL.

[0017] One aspect disclosed herein provides a pharmaceutical composition comprising a therapeutically effective amount of NMDP, formulated as a composition for intranasal administration, the composition comprising: a therapeutically effective amount of NMDP; a buffer; a penetration enhancer; and a surfactant.

[0018] In some embodiments, the composition further comprises one or more ingredients selected from vitamin E, benzyl alcohol, and dodecyl maltoside. In some embodiments, the concentration of NMDP in the administered pharmaceutical composition is 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37.5, 40, 42 .5, 45, 47.5, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 350, or 400 mg / ml. In some embodiments, the volume of the pharmaceutical composition administered is 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, or 1.0 ml. In some embodiments, the pharmaceutical composition is a sustained-release formulation. In some embodiments, the pharmaceutical composition is a sustained-release formulation. In some embodiments, the pharmaceutical composition is a controlled-release formulation. In some embodiments, the pharmaceutical composition releases in a continuous, variable, or pulsatile manner, or a combination thereof. In some embodiments, the pharmaceutical composition is in unit dosage form. In some embodiments, the unit dosage form is a dry powder, a semisolid, a mucoadhesive formulation, an intranasal vesicle unit, or a solution dosage form. In some embodiments, the composition is aqueous. In some embodiments, the composition is in the form of a gel or film. In some embodiments, the composition comprises micronized particles. In some embodiments, the unit weight of the unit dosage form is from about 10 mg to about 10 g. In some embodiments, the unit weight of the unit dosage form is from about 10 mg to about 50 mg, from about 10 mg to about 100 mg, from about 10 mg to about 150 mg, from about 10 mg to about 300 mg, from about 10 mg to about 500 mg, from about 10 mg to about 1 g, or from about 10 mg to about 5 g. In some embodiments, the unit dosage volume of the solution dosage form is less than about 900, 800, 700, 600, 500, 400, 300, 200, or 100 μL. In some embodiments, the concentration of the pharmaceutical composition comprising NMDP in the formulation is about 0.1% to about 20% by weight. In some embodiments, the pharmaceutical composition prolongs the residence time of the composition in the otic structure.In some embodiments, the formulation extends the residence time of the composition in the ear structure after a single dose by at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 8 days, or at least 14 days, at least 21 days, or at least 1 month, or at least 6 weeks. In some embodiments, the formulation increases the bioavailability of the composition in the ear structure. In some embodiments, the formulation increases the steady-state level of the composition in the ear structure. In some embodiments, the formulation increases the time to reach the Cmax of the therapeutic concentration that can alleviate the symptoms of hearing impairment in a subject in need. In some embodiments, the formulation extends the time that the concentration of the composition remains above the minimum therapeutic concentration (i.e., Cmin) necessary to alleviate the symptoms of hearing impairment in a subject in need. In some embodiments, the concentration of the composition in the ear structure remains at a concentration greater than Cmin or approximately greater than Cmin over a period of at least 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days, 3 weeks, or 1 month. In some embodiments, the pharmaceutical composition further comprises a second pharmaceutically active agent.

[0019] Aspects disclosed herein provide a pharmaceutical composition comprising a therapeutically effective amount of NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a therapeutically effective amount of NMDP; a solvent; and water. In some embodiments, the solvent is PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof. In some embodiments, the solvent comprises at least 0.00001% and up to 94% by weight of the composition. In some embodiments, the solvent comprises at least 66% and up to 94% by weight of the composition. In some embodiments, water comprises at least 0.00001% and up to 20% by weight of the composition. In some embodiments, the composition further comprises a first solvent and a second solvent. In some embodiments, the first solvent comprises mPEG and the second solvent comprises PEG. In some embodiments, the mPEG comprises mPEG350. In some embodiments, the PEG comprises PEG400. In some embodiments, the therapeutically effective amount of NMDP comprises at least 10 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / mL, at least 30 mg / mL, at least 35 mg / mL, at least 40 mg / mL, at least 45 mg / mL, at least 50 mg / mL, at least 55 mg / mL, at least 60 mg / mL, at least 65 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 95 mg / mL, at least 100 mg / mL, at least 110 mg / mL, or greater than 110 mg / mL. In some embodiments, mPEG comprises about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, or about 80% to about 90% by weight in the composition, PEG comprises about 10% to about 50%, about 20% to about 50%, about 30% to about 50%, or about 40% to about 50% by weight in the composition, and water comprises about 5% to about 20%, about 10% to about 20%, or about 15% to about 20% by weight in the composition. In some embodiments, the composition further comprises ethanol. In some embodiments, ethanol comprises about 1% to about 10%, about 2% to about 10%, about 3% to about 10%, about 4% to about 10%, about 5% to about 10%, about 6% to about 10%, about 7% to about 10%, about 8% to about 10%, or about 9% to about 10% in the composition.

[0020] These and other objects and features of the present disclosure will be more fully understood when the following detailed description of the present disclosure is read in conjunction with the accompanying drawings and examples.

[0021] Incorporated by Reference

[0022] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The novel features of the present disclosure are particularly set forth in the appended claims. The features and advantages of the present disclosure may be better understood by reference to the following detailed embodiments, which illustrate the principles of the present disclosure, in which the principles of the present disclosure are used, and the accompanying drawings:

[0024] Figure 1A Results from a no-go trial are presented, demonstrating the efficacy of different doses of the target medication for treating tinnitus compared with subjects treated with saline;

[0025] Figure 1B The results of the go (execution) test are shown, which indicate the subjects' motivation, listening, and learning and memory abilities, and serve as a control for the no-go test, used to evaluate the effects of drugs on the subjects' behavior;

[0026] Figure 2A It is a chart summarizing relevant results, showing the subjects' motivation, listening, and learning and memory abilities, and serves as a control for the no-go test to evaluate the effects of drugs on the subjects' behavior;

[0027] Figure 2B A graph summarizing the relevant results is shown, indicating the subjects' motivation, hearing, and learning and memory abilities, and serving as a control for the no-go test to assess the effects of the drug on the subjects' behavior;

[0028] Figure 2C A graph summarizing the results of the no-go trial showing the efficacy of different doses of the target drug for the treatment of tinnitus or tinnitus-like symptoms compared with subjects treated with saline is shown;

[0029] Figure 2D A graph summarizing the results of a no-go trial showing the efficacy of different doses of a target drug for the treatment of tinnitus or tinnitus-like symptoms compared with subjects treated with saline is shown;

[0030] Figure 3 The go / no-go test method based on sound avoidance detection used in this paper is summarized. DETAILED DESCRIPTION

[0031] NMDP is a calcium channel blocker (CCB) belonging to the dihydropyridine class. It is a highly lipid-soluble substance that can rapidly cross the blood-brain barrier.

[0032] Formulating NMDP into a formulation suitable for intranasal delivery is challenging due to its high lipid solubility, including its low number of hydrogen bond donors and high number of hydrogen bond acceptors. NMDP molecules have few hydroxyl groups and instead contain many carbonyl groups, which are unable to donate electron pairs to form hydrogen bonds, thus reducing their water solubility.

[0033] In addition, in order to facilitate the intranasal administration of NMDP, an effective amount of NMDP should be dissolved or finely dispersed in a small volume of liquid carrier. Larger volumes of liquid will be discharged from the anterior nares or flow back to the pharynx, causing excess liquid to be swallowed. Therefore, if the administration volume is too large, some NMDP may be lost from the absorption site, making it difficult (or even impossible) to administer the correct dose of the therapeutic agent in a reproducible manner. Therefore, it is ideal to disperse a high concentration of dissolved NMDP or NMDP components in a small volume of liquid carrier for intranasal administration.

[0034] Furthermore, to support drug absorption through the nasal mucosa, two natural protective functions must be bypassed: mucociliary clearance (MCC) and the tissue's barrier properties. Ideally, an effective dose would be able to bypass the natural properties of the mucus layer as a protective layer (i.e., enhancing drug absorption) and bypass MCC as an effective cleaning mechanism. In other words, the residence time of the administered dose on the mucus layer needs to be prolonged.

[0035] Given the unmet need for pharmaceutical compositions for treating hearing impairment in the prior art, the present invention discloses a pharmaceutical composition for treating hearing impairment (e.g., tinnitus or Meniere's disease). The present disclosure relates to a novel pharmaceutical formulation for delivering an effective dose of NMDP via mucosal delivery (e.g., intranasal delivery) to treat hearing impairment.

[0036] It should be understood that mucosal delivery has many advantages compared to conventional drug formulation administration methods (such as intravenous administration, injection or oral administration). For example, patients do not need to arrange and remember to take pills, or suffer painful injections. Administration is not affected by stomach or digestive problems. Mucosal delivery enables drugs to avoid degradation in the gastrointestinal tract or liver. Therefore, mucosal delivery is particularly meaningful for molecules with limited systemic bioavailability and short half-life (such as NMDP). In addition, mucosal delivery can quickly deliver active agents to subjects, thereby quickly alleviating the symptoms of hearing disorders such as tinnitus or Meniere's disease.

[0037] Mucosal administration (e.g., intranasal, buccal, sublingual, rectal, and pulmonary) is receiving particular attention because it avoids many of the disadvantages of injectable therapeutics while still providing a strong and rapid systemic effect. To be an attractive alternative to injection, mucosal administration (e.g., intranasal) should not cause significant pain, discomfort, or irritation, nor should it cause any irreversible damage to the mucosal surface. However, in the case of acute health-threatening indications, relatively high local irritation to the mucosa may be acceptable.

[0038] In mucosal administration, such as intranasal, buccal, or rectal administration, the therapeutic agent should be administered to the mucosa in a carrier that allows it to penetrate or be absorbed through the mucosa. In order to penetrate mucus, the carrier should be biocompatible with mucus and therefore have a certain degree of hydrophilicity. However, the carrier should also preferably be lipophilic to solubilize clinically relevant amounts of the therapeutic agent of interest.

[0039] The extensive capillary network beneath mucosal surfaces, particularly in the nasal mucosa, is ideally suited to provide rapid and efficient systemic absorption of drugs, vaccines, and biologics. Furthermore, the nasal epithelium actually contains a single layer of epithelial cells (pseudostratified epithelium), making it more suitable for drug delivery than other mucosal surfaces with a squamous epithelial layer, such as the oral cavity and vagina.

[0040] Compared with other routes of administration, intranasal delivery has many advantages, namely its non-invasiveness, rapid achievement of therapeutically relevant concentrations in the blood, no first-pass metabolism, and ease of administration. A feasible intranasal delivery technology has the potential to enable drug developers to deliver approved products through new routes of administration, thereby creating innovative drugs. Compared with mucosal administration, intravenous administration of NMDP has several disadvantages, such as increased side effects. However, non-polar molecules (such as NMDP) are difficult to formulate into preparations suitable for intranasal delivery because they have low water solubility and a small nasal cavity volume, which limits the amount of solution that can be used to deliver the drug substance to the subject during intranasal administration. There is still a need in the prior art for NMDP intranasal preparations that can be used to treat hearing disorders such as tinnitus or Meniere's disease and can quickly relieve the symptoms of hearing disorders.

[0041] The present researchers have developed a composition that delivers an effective dose of an NMDP equivalent through the mucosa, which can relieve the symptoms of tinnitus or Meniere's disease after administration of one or more doses of the pharmaceutical composition.

[0042] definition

[0043] Unless otherwise defined, all technical and scientific terms or terminology used herein shall have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. In some embodiments, terms with commonly understood meanings are expressly defined herein to facilitate understanding and / or reference, and the inclusion of such definitions herein should not be construed as constituting a substantial difference from what is generally understood in the art.

[0044] Throughout this application, various embodiments may be presented in range format. It should be understood that descriptions in range format are provided for convenience and brevity only and should not be construed as immutable limitations on the scope of the present disclosure. Therefore, descriptions of ranges should be considered to have specifically disclosed all possible subranges within that range, as well as individual numerical values. For example, descriptions of ranges such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values within that range, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0045] As used in the specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a" includes a plurality of examples, including mixtures thereof.

[0046] The terms "determine," "measure," "evaluate," "assess," "measure," and "analyze" are generally used interchangeably herein to refer to forms of measurement. These terms include determining whether an element is present (e.g., detecting). These terms can include quantitative, qualitative, or both quantitative and qualitative determinations. An assessment can be relative or absolute. "Detecting the presence of" can include determining the amount of something present as well as determining whether it is present, depending on the context.

[0047] The terms "subject," "individual," or "patient" are generally used interchangeably herein. A "subject" can be a biological entity comprising expressed genetic material. The biological entity can be a plant, an animal, or a microorganism, including, for example, bacteria, viruses, fungi, and protozoa. A subject can be tissues, cells, and progeny of a biological entity obtained from vivo or cultured in vitro. The subject can be a mammal. The mammal can be a human. A subject can be diagnosed or suspected of being at high risk for a disease. In some embodiments, a subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0048] The term "in vivo" is used to describe events that occur within the body of a subject.

[0049] The term "ex vivo" is used to describe an event that occurs outside the body of a subject. An ex vivo assay is not performed on a subject, but rather on a sample that has been separated from the subject. An example of an ex vivo assay performed on a sample is an "in vitro" assay.

[0050] The term "in vitro" is used to describe events that occur within a container containing a laboratory reagent, thereby isolating it from the biological source from which the material was obtained. In vitro assays can include cell-based assays that use living or dead cells. In vitro assays can also include cell-free assays that do not use intact cells.

[0051] As used herein, the term "about" a numerical value refers to the numerical value plus or minus 10%. The term "about" a range refers to the range minus 10% of the lowest value and plus 10% of the highest value.

[0052] As used herein, the terms "treat" or "treatment" refer to a drug or other intervention regimen for obtaining a beneficial or desired result in the recipient. Beneficial or desired results include, but are not limited to, therapeutic benefit and / or prophylactic benefit. A therapeutic benefit may refer to the eradication or improvement of the symptoms or underlying condition being treated. In addition, a therapeutic benefit may be achieved when one or more physiological symptoms associated with the underlying condition are eradicated or improved, resulting in an improvement observed in the subject, even if the subject is still suffering from the underlying condition. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, arresting, or reversing the progression of a disease or condition, or any combination thereof. To obtain a prophylactic benefit, a subject at risk for a particular disease, or a subject who reports having one or more physiological symptoms of a disease, may receive treatment even if a diagnosis of the disease has not yet been made.

[0053] The term "NMDP" is intended to refer to NMDP or a pharmaceutically acceptable salt thereof. The term "equivalent to about ..." of NMDP is intended to refer to a specific volume, concentration, or amount of NMDP free base provided by a volume, concentration, or amount of an NMDP salt. Thus, the specific amount relates to the amount of NMDP free base, not the amount of an NMDP salt, even if such a salt is used in the composition. In one embodiment, the compositions, methods, and uses of the present disclosure include the use of NMDP citrate.

[0054] The term "formulation" is intended to refer to the selection of excipients, carriers, vehicles, preservatives, stabilizers, etc. when preparing a medicament using the composition. The term "formulation" is also intended to refer to the selection of a device for delivering the composition, or the selection of a containment device for administering or storing the composition.

[0055] The term "dosage unit" refers to a composition administered by a single delivery operation. In the embodiment in which the composition is formulated for mucosal administration by intranasal delivery, a dosage unit is the volume of the composition administered by a single delivery operation or the amount of the medicament administered. A delivery operation is the operation of delivering a dosage unit. In this embodiment, a delivery operation is a dosage unit administered to the nasal cavity by a delivery system (such as nasal spray or other means well known to those skilled in the art). Suitable devices are commercially available from, for example, Pfeiffer and Valois. The terms "dosage" and "therapeutic dose" refer to the total amount of the medicament administered by a dosage unit or the volume of the composition during treatment. Treatment refers to giving a composition during a single attack of hearing impairment (comprising tinnitus or Meniere's disease or the symptom of tinnitus or Meniere's disease), and the attack continues until the symptom relief of tinnitus or Meniere's disease or tinnitus or Meniere's disease.

[0056] The term "onset time" is intended to mean the moment at which a patient begins to experience relief from tinnitus or Meniere's disease, or symptoms of tinnitus or Meniere's disease, typically as a result of achieving adequate plasma concentrations of NMDP. Achieving adequate plasma concentrations in all patients varies depending on the patient, patient category, and the type and nature of the symptoms of tinnitus or Meniere's disease, or tinnitus or Meniere's disease experienced. The "onset" in "onset time" refers to relief from the symptoms of tinnitus or Meniere's disease, or tinnitus or Meniere's disease.

[0057] The term "duration of action" refers to the time that a patient experiences tinnitus or Meniere's disease, or relief from symptoms of tinnitus or Meniere's disease.

[0058] By "extended period of time" is meant a period of time exceeding about 4 months, preferably exceeding about 6 months.

[0059] The phrase "an amount effective to inhibit one or more symptoms of tinnitus or Meniere's disease" refers to a dose of a therapeutic compound that reduces one or more symptoms of tinnitus or Meniere's disease in a subject following delivery of the pharmaceutical composition.

[0060] The terms used herein are used only to describe specific embodiments and are not intended to limit the present disclosure. The terms "comprising" and "including" used in the claims should not be interpreted as being limited to the components and steps listed thereafter; they do not exclude other components or steps. They should be interpreted as indicating the presence of the stated features, integers, steps and / or components, but do not exclude the presence and / or addition of one or more other features, integers, steps or components or groups thereof. Thus, the scope of the expression "a composition comprising A and B" should not be limited to compositions consisting only of components A and B. Likewise, the scope of the expression "a method comprising steps X and Z" should not be limited to methods consisting only of those steps.

[0061] As used herein, the term "about" a numerical value refers to that numerical value plus or minus 10%. The term "about" a range refers to that range minus 10% of its lowest value and plus 10% of its highest value. For example, the term "about" can be immediately understood to mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the value. As an illustration, a numerical range of "about 1 to about 5" should be interpreted as including not only the explicitly listed values of about 1 to about 5, but also the individual values and subranges within the indicated range. This includes at least the degree of expected experimental error, technical error, and instrumental error for a given experiment, technique, or instrument used to measure the value.

[0062] As used herein, the term "and / or" includes any combination of one or more of the associated listed items. Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should also be understood that terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the specification and related fields, and should not be interpreted in an idealized or overly formal sense unless explicitly defined as such herein. For the sake of brevity and / or clarity, well-known functions or structures may not be described in detail.

[0063] It should be understood that when an element is referred to as being "on," "attached to," "connected to," "coupled to," "in contact with," etc., another element, it can be directly on, attached to, connected to, coupled to, or in contact with the other element, or intervening elements may be present. In contrast, when an element is referred to as being "directly on," "directly attached to," "directly connected to," "directly coupled to," or "directly in contact with" another element, for example, there are no intervening elements. Those skilled in the art will also understand that references to structures or features disposed "adjacent" another feature can have portions overlapping or beneath the adjacent feature.

[0064] The terms "active agent," "pharmaceutically active agent," "active ingredient," "API," "pharmaceutically active ingredient," "active substance," "active molecule," "active compound," or "drug" are used interchangeably with NMDP or a salt thereof.

[0065] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. It should also be understood that terms such as those defined in commonly used dictionaries should be interpreted as having the same meaning as in the specification and the relevant art, and should not be interpreted in an idealized or overly formal sense unless explicitly defined as such herein. For the sake of brevity and / or clarity, well-known functions or configurations may not be described in detail.

[0066] The disintegrator included in the disclosed compositions is an inert carrier, as described in detail below. It should be noted that "disintegrator," "carrier," "diluent," and "anti-caking agent" are used interchangeably herein to refer to an inert ingredient added to the pharmaceutical composition comprising the second type of particles.

[0067] "Mucosal delivery enhancers" are defined as chemicals and other excipients that, when added to a formulation comprising water, salts and / or conventional buffers and NMDP (control formulation), produce a formulation that significantly increases the transport of NMDP across the mucosa, as measured by maximum blood, serum or cerebrospinal fluid concentration (Cmax) or by the area under the concentration-time curve (AUC). Mucosal membranes include the nasal, oral, intestinal, buccal, bronchopulmonary, vaginal and rectal mucosal surfaces, and include all membranes that secrete mucus lining all body cavities or passages that communicate with the outside world. Mucosal delivery enhancers are sometimes referred to as carriers, excipients, additives, enhancers or promoters (including, for example, thickeners).

[0068] "Endotoxin-free formulation" refers to a formulation comprising NMDP and one or more mucosal delivery enhancers, which is substantially free of endotoxins and / or related pyrogenic substances. Endotoxins include toxins that are confined to the interior of microorganisms and are released only when the microorganisms decompose or die. Pyrogenic substances include pyrogenic and heat-resistant substances (glycoproteins) from the outer membranes of bacteria and other microorganisms. If administered to humans, these substances can cause fever, hypotension, and shock. The production of endotoxin-free formulations may require special equipment, professional and technical personnel, and may be much more expensive than the production of non-endotoxin-free formulations.

[0069] "Non-infusion administration" refers to any method of delivery that does not involve direct injection into an artery or vein, i.e., a method of forcing or driving (usually a fluid) into something, especially methods introduced into a body part by a needle, syringe, or other invasive method. Non-infusion administration includes subcutaneous injection, intramuscular injection, intraperitoneal injection, and non-injection delivery methods to mucous membranes.

[0070] According to some embodiments, a "subject" is a subject in which symptoms and signs, physical examination results and / or psychological test results are determined and recorded for an individual's condition (i.e., a disease or condition). As used herein, the disease or condition is hearing impairment. As used herein, the disease or condition is tinnitus or Meniere's disease. As used herein, a "subject" is intended to be a human subject, but is not necessarily limited thereto. The subject can be male or female, and can be of any race or ethnicity, including but not limited to Caucasians, African Americans, Africans, Asians, Hispanics, Native Americans, etc. As used herein, a subject is an animal, especially a mammal, such as a dog, cat, cow, goat, horse, sheep, pig, rodent (such as rats and mice), rabbit, primate (including non-human primates), etc., which can be treated according to the methods of the present disclosure or screened for veterinary and pharmaceutical or drug development purposes. According to some embodiments of the present disclosure, the subject includes a patient or human being who needs to be treated for a condition that can be treated by ...

[0071] NMDP composition

[0072] The pharmaceutical compositions of the present disclosure comprise NMDP or a salt thereof in a concentration equivalent to about 0.4 to 75 mg / mL of NMDP in a suitable solvent. In some embodiments, the equivalent concentration of NMDP may be greater than 75 mg / mL. In some embodiments, the equivalent concentration of NMDP may be 25 mg / mL. In some embodiments, the equivalent concentration of NMDP may be 50 mg / mL. In some embodiments, the equivalent concentration of NMDP may be 75 mg / mL. In some embodiments, the equivalent concentration of NMDP may be 100 mg / mL. The composition is suitable for formulation for mucosal administration, typically for delivering NMDP through the nasal mucosa.

[0073] The therapeutically effective amount of NMDP or a salt thereof may be at least 1 μg of NMDP or a salt thereof per kilogram of subject. The therapeutically effective amount of NMDP or a salt thereof may also be less than 100 mg of NMDP or a salt thereof per kilogram of subject. The therapeutically effective amount of NMDP or a salt thereof ranges from 1 μg to 1000 mg of NMDP or a salt thereof per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof ranges from 16 mg to 24 mg of NMDP or a salt thereof per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof ranges from 30 mg to 100 mg of NMDP or a salt thereof per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof ranges from 30 mg to 100 mg of NMDP or a salt thereof per kilogram of subject.

[0074] The therapeutically effective amount of NMDP or its salt can be administered before the onset of hearing impairment (such as tinnitus or Meniere's disease).The therapeutically effective amount of NMDP or its salt can be administered at least about 1 hour before the onset of hearing impairment, such as at least about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks or 1 month before the onset of hearing impairment.The therapeutically effective amount of NMDP or its salt can be administered less than about 1 month before the onset of hearing loss, such as less than about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks or 1 month before the onset of hearing impairment.In some embodiments, the therapeutically effective amount of NMDP or its salt is administered at least 12 hours before the onset of hearing impairment. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is administered less than 10 days before the onset of hearing impairment. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is administered between 12 hours and 10 days before the onset of hearing impairment. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is administered between 12 hours and 48 hours before the onset of hearing impairment.

[0075] The treatment effective amount of NMDP or its salt can be administered after or during the onset of hearing impairment (such as tinnitus or Meniere's disease).The treatment effective amount of NMDP or its salt can be administered at least about 1 minute after the onset of hearing impairment, such as at least about 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 12 minutes, 24 minutes, 36 minutes, 48 minutes, 54 minutes or 60 minutes after the onset of hearing impairment.The treatment effective amount of NMDP or its salt can be administered at least about 1 hour after the onset of hearing impairment, such as at least about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks or 1 month after the onset of hearing impairment. A therapeutically effective amount of NMDP or a salt thereof can be administered less than about 1 month after the onset of hearing loss, for example, less than about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks or 1 month after the onset of hearing impairment. In some embodiments, a therapeutically effective amount of NMDP or a salt thereof is administered at least 12 hours after the onset of hearing impairment. In some embodiments, a therapeutically effective amount of NMDP or a salt thereof is administered less than 10 days after the onset of hearing impairment. In some embodiments, a therapeutically effective amount of NMDP or a salt thereof is administered between 12 hours and 10 days after the onset of hearing impairment. In some embodiments, a therapeutically effective amount of NMDP or a salt thereof is administered between 12 hours and 48 hours after the onset of hearing impairment.

[0076] Effective dose range

[0077] The therapeutically effective amount of NMDP can depend on the subject's weight. In some embodiments, the therapeutically effective amount of NMDP is at least about 1 mg of NMDP per kilogram of subject, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200 mg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP is less than about 1000 μg of NMDP per kilogram of subject, for example, less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 μg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 1 μg to 1000 μg of NMDP per kilogram of subject, such as about 1-700, 1-500, 1-300, 1-100, 1-50, 1-10, 10-700, 10-500, 10-300, 10-100, 10-80, 10-60, 10-40, 10-20, 50-700, 50-500, 50-300, 50-100, 100-700, 100-500, 100-300, 300-700, 300-500, or 500-700 μg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 1 μg to 10 μg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 10 μg to 100 μg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 100 μg to 500 μg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 1 μg to 1000 mg of NMDP per kilogram of subject, such as about 1-700, 1-500, 1-300, 1-100, 1-50, 1-10, 10-700, 10-500, 10-300, 10-100, 10-80, 10-60, 10-40, 10-20, 50-700, 50-500, 50-300, 50-100, 100-700, 100-500, 100-300, 300-700, 300-500, or 500-700 mg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 16 mg to 24 mg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 30 mg to 100 mg of NMDP per kilogram of subject. In some embodiments, the therapeutically effective amount of NMDP ranges from about 50 mg to 140 mg of NMDP per kilogram of subject.In some embodiments, the therapeutically effective amount of NMDP ranges from about 115 mg to 125 mg of NMDP per kilogram of the subject.The therapeutically effective amount of NMDP can also be the daily NMDP dose for the subject.

[0078] In pharmaceutical compositions comprising a therapeutically effective amount of an NMDP and one or more APIs, the amount of API in the pharmaceutical composition can depend on the weight of the subject. In some embodiments, the amount of API in the pharmaceutical composition is at least about 1 mg of API per kilogram of subject, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200 mg of API per kilogram of subject. In some embodiments, the amount of API in the pharmaceutical composition is less than about 1 mg of API per kilogram of subject, for example, less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200 mg of API per kilogram of subject. In some embodiments, the amount of API in the pharmaceutical composition ranges from about 1 to 200 mg of API per kilogram of subject. In some embodiments, the therapeutically effective amount of the API ranges from about 30 mg to 100 mg of API per kilogram of subject. In some embodiments, the therapeutically effective amount of the API ranges from about 50 mg to 140 mg of API per kilogram of subject. The therapeutically effective amount of the API can also be the daily dose of the API for the subject. For example, the daily dose of a sodium channel blocker, antioxidant, NMDA antagonist, SSRI, or SSRI / NMDA antagonist combination can be about 1 to 500 mg / day, preferably 4 to 250 mg / day.

[0079] The concentration of NMDP in the pharmaceutical composition can be at least about 0.1%, for example, at least about 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% by weight. The concentration of NMDP in the pharmaceutical composition can be less than about 99%, for example, less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 99% by weight. The concentration range of NMDP in the pharmaceutical composition can be about 0.1 to 99% by weight, for example, 0.1 to 0.5%, 0.1 to 1%, 0.5 to 1%, 1 to 2%, 1 to 5%, 1 to 10%, 2 to 5%, 2 to 10%, 5 to 10%, 10 to 15%, 15 to 20%, 20 to 30%, 30 to 50%, 50 to 70%, 70 to 90%, 90 to 95% or 95 to 99%. In some embodiments, the concentration range of NMDP in the pharmaceutical composition is 1 to 10%. In some embodiments, the concentration range of NMDP in the pharmaceutical composition is 10 to 50%. In some embodiments, the concentration range of NMDP in the pharmaceutical composition is 70 to 90%.

[0080] The concentration of the API in the pharmaceutical composition can be at least about 0.1%, for example, at least about 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% by weight. The concentration of the API in the pharmaceutical composition can be less than about 99%, for example, less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 99% by weight. The concentration of the API in the pharmaceutical composition can range from about 0.1 to 99% by weight, for example, 0.1 to 0.5%, 0.1 to 1%, 0.5 to 1%, 1 to 2%, 1 to 5%, 1 to 10%, 2 to 5%, 2 to 10%, 5 to 10%, 10 to 15%, 15 to 20%, 20 to 30%, 30 to 50%, 50 to 70%, 70 to 90%, 90 to 95%, or 95 to 99%. In some embodiments, the concentration of the API in the pharmaceutical composition ranges from 1 to 10%. In some embodiments, the concentration of the API in the pharmaceutical composition ranges from 10 to 50%. In some embodiments, the concentration of the API in the pharmaceutical composition ranges from 70 to 90%.

[0081] The therapeutically effective amount of NMDP can be in the form of a dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit, or solution. The unit weight of the dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit, or solution can be at least about 1 mg, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg. The unit weight of the dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit, or solution can be at least about 1 g, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 g. The unit weight of the dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit or solution dosage form may be less than about 1000 mg, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800 or 900 mg. The unit weight of the dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit or solution dosage form may be less than about 100 g, for example, less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 g. The unit weight of the dry powder, semisolid, mucoadhesive formulation, intranasal vesicle unit, or solution dosage form can range from about 1 mg to 10 g, for example, from about 1 mg to 10 mg, 10 mg to 50 mg, 50 mg to 100 mg, 100 mg to 200 mg, 200 mg to 400 mg, 400 mg to 600 mg, 600 mg to 800 mg, 800 mg to 1 g, 1 g to 2 g, 2 g to 5 g, or 5 g to 10 g. The therapeutically effective amount of NMDP can be in the form of a solution. The unit volume of the solution dosage form can be at least about 1 mL, for example, at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mL. The unit volume of the solution dosage form can be less than about 1000 mL, for example, less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mL.The unit volume of the solution dosage form can range from about 1 to 500 mL, for example, in the range of 1-500, 1-300, 1-100, 1-80, 1-60, 1-40, 1-20, 1-10, 1-5, 10-500, 10-300, 10-100, 10-80, 10-60, 10-40, 10-20, 20-500, 20-300, 20-100, 20-80, 20-60, 20-40, 40-500, 40-300, 40-100, 40-80, 40-60, 60-500, 60-300, 60-100, 60-80, 80-500, 80-300, 80-100, 100-500, 100-300 or 300-500 mL.

[0082] Combination therapy

[0083] The therapeutically effective amount of NMDP or a salt thereof can be the sole active pharmaceutical ingredient (API). Alternatively, the therapeutically effective amount of NMDP or a salt thereof can be used in combination with one or more additional APIs. Disclosed herein are medicaments, compositions, kits, and methods for preventing or treating hearing impairment by administering NMDP or a salt thereof (e.g., a therapeutically effective amount of NMDP or a salt thereof) and one or more active pharmaceutical ingredients (APIs, e.g., a therapeutically effective amount of an API).

[0084] The one or more APIs may include one or more sodium channel blockers, antioxidants, spin traps, N-methyl-D-aspartate (NMDA) antagonists, selective serotonin reuptake inhibitor (SSRI) antagonists, dopamine releasing agents (DRAs), acetylcholine release inducers, norepinephrine reuptake inhibitors (NERIs), monoamine oxidase-A inhibitors (MAIs), serotonin reuptake inhibitors (SRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), norepinephrine selective reuptake inhibitors (NSRIs), serotonin reuptake inhibitors (5HT SRIs), zonisamide, gabapentin, cannabinoids, or any combination thereof.

[0085] Also disclosed herein are drugs, compositions, kits, and methods for treating or preventing hearing impairment in subjects in need thereof, comprising administering a therapeutically effective amount of an NMDP or a salt thereof and one or more active pharmaceutical ingredients (APIs). The one or more APIs can be administered in a therapeutically effective amount. The one or more APIs can include one or more sodium channel blockers, antioxidants, spin traps, N-methyl-D-aspartate (NMDA) antagonists, selective serotonin reuptake inhibitors (SSRI) antagonists, dopamine releasers (DRAs), acetylcholine release inducers, norepinephrine reuptake inhibitors (NERIs), monoamine oxidase-A inhibitors (MAIs), serotonin reuptake inhibitors (SRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), norepinephrine selective reuptake inhibitors (NSRIs), serotonin reuptake inhibitors (5HT SRIs), zonisamide, gabapentin, cannabinoids, or any combination thereof.

[0086] The one or more APIs may include one or more antioxidants or spin traps. For example, the one or more antioxidants or spin traps may include allopurinol, glutathione, L-carnitine, methionine, or any combination thereof. The one or more APIs may include one or more NMDA antagonists. For example, the one or more NMDA antagonists may include riluzole, caroverine, memantine, magnesium, or any combination thereof. The one or more APIs may include one or more SSRI antagonists. For example, the one or more SSRI antagonists may include fluoxetine, sertraline, S-citalopram, alproclofen, or any combination thereof. The one or more APIs may include one or more DRAs. For example, the one or more DRAs may include amantadine. The one or more APIs may include one or more acetylcholine release inducers or NERIs. For example, the one or more acetylcholine release inducers or NERIs may include bifennilan. The one or more APIs may include one or more MAIs or SRIs. For example, the one or more MAIs or SRIs may include pyrindol. The one or more APIs may include one or more SNRIs. For example, the one or more SNRIs may include milnacipran, bicyclosporine, or both. The one or more APIs may include one or more CCBs. For example, the one or more CCBs may include NMDP, verapamil, or both. The one or more APIs may include one or more NSRIs. For example, the one or more NSRIs may include atomoxetine. The one or more APIs may include one or more 5-TH SRIs. For example, the one or more 5-TH SRIs may include indolizumab. The one or more APIs may include zonisamide.

[0087] The one or more APIs may include one or more drugs, such as gabapentin. Other drugs that may be used include anticonvulsants. Other drugs that may be used include drugs that stimulate gamma-aminobutyric acid (GABA) receptors. The one or more APIs may include cannabinoids. For example, the cannabinoid may be cannabis or any cannabis extract or a synthetic composition that stimulates cannabinoid receptors, CB1 receptors, CB2 receptors, or G-coupled receptors. These drugs may be used in combination with nimodipine or a salt thereof.

[0088] Reagent test kit

[0089] Disclosed herein is a kit for treating or preventing hearing impairment in a subject in need thereof, comprising an NMDP or a salt thereof and written instructions for using the NMDP or a salt thereof to treat or prevent hearing impairment. The NMDP or a salt thereof can be a therapeutically effective amount of the NMDP or a salt thereof. The therapeutically effective amount of the NMDP or a salt thereof can be in the form of a tablet, capsule, caplet, spray, powder, gel cap, powder, or solution. For example, the therapeutically effective amount of the NMDP or a salt thereof can be in the form of a powder. The kit can also comprise a sterile solution. The kit can also comprise a sterile solution to be mixed with the powder formulation prior to administering the therapeutically effective amount of the NMDP or a salt thereof. The kit can also comprise one or more active pharmaceutical ingredients (APIs). The one or more APIs may comprise one or more antioxidants, spin traps, N-methyl-D-aspartate (NMDA) antagonists, selective serotonin reuptake inhibitor (SSRI) antagonists, dopamine releasing agents (DRAs), acetylcholine release inducers, norepinephrine reuptake inhibitors (NERIs), monoamine oxidase-A inhibitors (MAIs), serotonin reuptake inhibitors (SRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), norepinephrine selective reuptake inhibitors (NSRIs), serotonin reuptake inhibitors (5HT SRIs), zonisamide, gabapentin, cannabinoids, or any combination thereof.

[0090] Molecular mechanism

[0091] Disclosed herein are methods for modulating a calcium signaling pathway in a subject in need thereof, comprising administering a therapeutically effective amount of an NMDP or a salt thereof. The calcium signaling pathway can be modulated by inhibiting calcium influx through voltage-gated calcium channels, α2-adrenergic receptor-regulated calcium channels, or both. The calcium signaling pathway can be modulated by inhibiting the release of cytoplasmic calcium stores, for example, in corpus cavernosum smooth muscle cells. The calcium signaling pathway can be modulated by inhibiting the production of tumor necrosis factor-α, for example, induced by calcium influx. The calcium signaling pathway can be modulated by blocking voltage-gated calcium channels. The calcium signaling pathway can be modulated by affecting the function of calcium-ATPase, calcium release channels, or both. The calcium signaling pathway can be modulated by releasing intracellular calcium, blocking calcium influx, or both. The calcium signaling pathway can be modulated by blocking endosomal calcium channels, such as two-pore channels. The calcium signaling pathway can be modulated by blocking T-type and / or L-type calcium currents. The calcium signaling pathway can be modulated by inhibiting the activity of BKCa channels. The calcium signaling pathway can be modulated by inhibiting calcium release-activated channels.

[0092] Disclosed herein are methods for modulating an oxidative pathway in a subject in need thereof, comprising administering a therapeutically effective amount of NMDP or a salt thereof. The oxidative pathway can be modulated by reducing oxidative stress, downregulating miRNA-155, reducing TNF-α in the NF-κB signaling pathway, or a combination thereof. The oxidative pathway can be modulated by scavenging free radicals. The oxidative pathway can be modulated by reducing peroxide levels. The oxidative pathway can be modulated by regulating cellular redox state. The oxidative pathway can be modulated by inhibiting the formation of reactive oxygen species, inhibiting the upregulation of Mac-1, neutrophil adhesion to fibrinogen, or any combination thereof. The oxidative pathway can be modulated by preventing hydrogen peroxide-induced oxidative neuronal cell damage.

[0093] Disclosed herein are methods for modulating an anti-inflammatory pathway in a subject in need thereof, comprising administering a therapeutically effective amount of an NMDP or a salt thereof. The anti-inflammatory pathway can be modulated by reducing tube formation during angiogenesis. For example, reducing tube formation during angiogenesis can inhibit the post-receptor pathway of IL-1α and / or platelet-derived growth factor-BB in chronic inflammation. The anti-inflammatory pathway can be modulated by inhibiting IL-1 and / or TNF-induced leukocyte infiltration into the air sac. The anti-inflammatory pathway can be modulated by inhibiting the synthesis of prostaglandin E. The anti-inflammatory pathway can be modulated by inhibiting IL-1, TNF-α, IL-6, IL-8, IgG, neutrophil phagocytosis, or any combination thereof. For example, the IL-1, TNF-α, IL-6, and / or IL-8 can be derived from monocytes, and the IgG can be derived from B cells. The anti-inflammatory pathway can be modulated by inhibiting the increase in lipopolysaccharide-induced secretion of TNF-α, IL-1β, and / or high-mobility group protein B1 by peritoneal macrophages.

[0094] Patient selection

[0095] In one aspect, the present invention provides a method for selecting a subject in need for treatment of hearing impairment or hearing impairment symptoms, comprising: administering an L-type calcium channel blocker to the subject; if the subject responds to the L-type calcium channel blocker, selecting the patient for treatment of hearing impairment or hearing impairment symptoms; and if the subject responds to the L-type calcium channel blocker, administering a therapeutically effective amount of NMDP or a salt thereof to the subject. In some embodiments, the L-type calcium channel blocker comprises carbamazepine. In some embodiments, the patient exhibits hearing loss. In some embodiments, the patient exhibits one or more symptoms associated with: a) hearing loss; b) changes in auditory speech recognition as measured by a speech in noise test; c) changes in auditory speech recognition as measured by a digit in noise test; d) changes in low-frequency hearing thresholds; e) changes in the severity of tinnitus or Meniere's disease; g) changes in the loudness of tinnitus or Meniere's disease; h) changes in the severity of vertigo; i) changes in the sensation of fullness in the ears; j) changes in dizziness; and k) changes in hair cell function as observed by changes in ABR thresholds.

[0096] In one aspect, the present invention provides a method for selecting a subject in need for treatment of tinnitus or Meniere's disease, or a symptom of tinnitus or Meniere's disease, comprising: administering an L-type calcium channel blocker to the subject; if the subject responds to the L-type calcium channel blocker, selecting the subject for treatment of tinnitus or Meniere's disease, or a symptom of tinnitus or Meniere's disease; and if the subject responds to the L-type calcium channel blocker, administering a therapeutically effective amount of NMDP or a salt thereof to the subject. In some embodiments, the L-type calcium channel blocker comprises carbamazepine.

[0097] Condition to be treated

[0098] The hearing impairment of the subject in need may be NIHL, ARHL, hearing loss caused by drugs or injury, or tinnitus or Meniere's disease. The hearing impairment of the subject may also include a combination of two or all of the four diseases listed. For example, tinnitus or Meniere's disease and drug-induced hearing loss may be caused by ototoxic drugs. The ototoxic drugs may include chemotherapy drugs, anti-tumor drugs, antibiotics, loop diuretics, quinine or quinine compounds, or salicylates or salicylates. In some embodiments, the ototoxic drug is not streptomycin. In some embodiments, the antibiotic is not streptomycin. In some embodiments, the hearing impairment is not caused by streptomycin.

[0099] Disclosed herein are medicaments, compositions, kits, and methods for preventing and / or treating hearing disorders, including, but not limited to, various conditions such as noise-induced hearing loss (NIHL), age-related hearing loss (ARHL or presbycusis), drug- or injury-induced hearing loss, central auditory dysfunction (CAPD), tinnitus, or Meniere's disease.

[0100] NIHL - Noise-Induced Hearing Loss

[0101] NIHL is one of the most significant health hazards in both occupational and recreational settings. However, there are currently no FDA-approved medications to mitigate NIHL, and the development of effective treatments is hampered by the complex cellular and molecular pathways involved. NIHL can cause a range of damage, from hair cell fatigue to cell death in the ear. Therefore, NIHL likely results from overstimulation of hair cells and supporting structures. Structural damage to hair cells, primarily outer hair cells, can lead to hearing loss, characterized by attenuation and distortion of incoming auditory stimuli.

[0102] NIHL can be caused by a single exposure to excessive noise. For example, exposure to sounds exceeding 80, 90, 100, 110, 120, 130, 140, or 150 decibels over a short period of time can lead to NIHL. Alternatively, NIHL can be caused by repeated exposure to noise over a period of time. For example, exposure to sounds exceeding 60, 65, 70, 75, 80, 85, 90, 95, or 100 dB for more than eight hours per day can lead to NIHL. Symptoms of NIHL can include tinnitus or Meniere's disease, ear pain, hyperacusis, dizziness, vertigo, and / or vestibular damage in the inner ear.

[0103] Disclosed herein are medicaments, compositions, kits, and methods for treating or preventing hearing loss (e.g., NIHL) in a subject in need thereof, wherein the methods comprise administering a therapeutically effective amount of NMDP or a salt thereof. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reduce or eliminate NIHL. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to prevent or prevent NIHL. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reverse NIHL or at least partially restore hearing. The subject may be at risk of developing hearing loss (e.g., NIHL) or the subject may be experiencing hearing loss (e.g., NIHL).

[0104] ARHL or presbycusis

[0105] Age-related hearing loss (ARHL), or presbycusis, is a major health problem for which there is currently no treatment or preventive measure. Age-related hearing loss develops gradually over time and is barely noticeable to affected individuals in its early stages. The cause of ARHL, or presbycusis, is generally believed to be a degeneration of the auditory nervous system, specifically the auditory nerve in the ear. It is the most common form of hearing loss in people over 55 years of age.

[0106] Early noise damage may be the cause of ARHL or presbycusis. The subject may be suffering from or at risk of hearing loss (e.g., ARHL or presbycusis). For example, the subject may be suffering from or at risk of hearing loss (e.g., ARHL or presbycusis) at least 30 years old, 35 years old, 40 years old, 45 years old, 50 years old, 55 years old, 60 years old, 65 years old, 70 years old, 75 years old, 80 years old, 85 years old, 90 years old, 95 years old or 100 years old. In some embodiments, the subject may be 50 years old.

[0107] Disclosed herein are medicines, compositions, kits and methods for treating or preventing hearing loss (e.g., ARHL or presbycusis) in a subject in need thereof, wherein the method comprises administering a therapeutically effective amount of NMDP or a salt thereof. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to alleviate or eliminate ARHL or presbycusis. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to prevent or prevent ARHL or presbycusis. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reverse ARHL or presbycusis or at least partially restore hearing. The subject may be at risk of hearing loss (e.g., ARHL or presbycusis), or the subject may be suffering from hearing loss (e.g., ARHL or presbycusis).

[0108] Traumatic brain injury and subarachnoid hemorrhage

[0109] More than half of patients with traumatic brain injury (TBI) experience tinnitus, and more than 20% of patients with subarachnoid hemorrhage (SAH) experience hearing loss. Therefore, in certain aspects, the compositions and methods disclosed herein can be used to treat hearing loss induced by TBI and / or SAH. In some embodiments, the NMDP formulations disclosed herein can be used to treat hearing loss (such as tinnitus) induced by TBI and / or SAH.

[0110] Drug-induced hearing loss

[0111] Ototoxic drugs, such as chemotherapeutic agents, antineoplastic drugs, antibiotics, loop diuretics, quinine or quinine-like compounds, and salicylates or salicylate-like compounds, can cause drug-induced hearing loss. For example, aminoglycoside antibiotics have been used to treat Gram-negative and certain aerobic Gram-positive infections. However, despite their usefulness, they are associated with serious side effects, including ototoxicity associated with the destruction of sensory hair cells in the organ of Corti in the inner ear. In addition, surgery near or on the auditory nerve can lead to hearing loss, which can subsequently cause tinnitus or tinnitus-like symptoms.

[0112] Disclosed herein are medicaments, compositions, kits, and methods for treating or preventing hearing loss (e.g., injury- or drug-induced hearing loss) in a subject in need thereof, wherein the method comprises administering a therapeutically effective amount of NMDP or a salt thereof. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reduce or eliminate such hearing loss. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to prevent or prevent such hearing loss. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is not only sufficient to prevent drug-induced hearing loss, but also to at least partially restore hearing. The subject may be at risk of developing hearing loss (e.g., drug-induced hearing loss), or the subject may be suffering from hearing loss (e.g., drug-induced hearing loss). In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is not only sufficient to prevent drug-induced hearing loss, but also to act synergistically with anticancer drugs to kill cancer cells.

[0113] The therapeutically effective amount of NMDP or its salt can prevent the occurrence of damage or drug-induced hearing loss. For example, disclosed herein is a method for preventing drug-induced hearing loss, including administering a therapeutically effective amount of NMDP or its salt to a subject in need thereof before administering one or more ototoxic drugs for treating non-hearing loss conditions. Administration of a therapeutically effective amount of NMDP or its salt can begin at most about 12 months (e.g., 1 day to 60 days) before administering one or more ototoxic drugs. Administration of a therapeutically effective amount of NMDP or its salt can also begin on the same day as administering one or more ototoxic drugs. In some embodiments, administration of a therapeutically effective amount of NMDP or its salt can begin at most about 12 months (e.g., 1 hour, 6 hours, 12 hours, 24 hours, 2 days, 4 days, 6 days, 8 days, 10 days, 20 days, 1 month, 2 months, 4 months, 6 months, 8 months, 10 months, or 12 months) before administering one or more ototoxic drugs. In some embodiments, administration of a therapeutically effective amount of NMDP or a salt thereof can begin 1 day to 12 months (e.g., 1 day to 2 days, 2 days to 10 days, 10 days to 1 month, 1 month to 3 months, 3 months to 6 months, or 6 months to 12 months) prior to administration of one or more ototoxic drugs.

[0114] The therapeutically effective amount of NMDP or a salt thereof can treat or prevent drug-induced hearing loss, such as improving drug-induced hearing loss, alleviating or eliminating tinnitus or Meniere's disease, partially or completely restoring hearing, or preventing further hearing loss caused by the ototoxic effects of one or more ototoxic drugs. The methods disclosed herein provide a regimen for administering a pharmaceutical composition in response to a significant decrease in hearing function that occurs or develops during administration of one or more ototoxic drugs.

[0115] The NMDP or its salt of administering a therapeutically effective amount can be continued during the administration of one or more ototoxic drugs. The NMDP or its salt of administering a therapeutically effective amount can be stopped on the same day that one or more ototoxic drugs stop administration. The NMDP or its salt of administering a therapeutically effective amount can at least continue about 1 day (for example, at least continue about 1 day, 2 days, 4 days, 6 days, 8 days, 10 days, 20 days, 1 month, 2 months, 4 months, 6 months, 8 months, 10 months or 12 months) after one or more ototoxic drugs stop administration. In some embodiments, the NMDP or its salt of administering a therapeutically effective amount can at least continue about 1 day to 12 months (for example, 1 day to 2 days, 2 days to 10 days, 10 days to 1 month, 1 month to 3 months, 3 months to 6 months or 6 months to 12 months) after one or more ototoxic drugs stop administration.

[0116] Examples of ototoxic drugs include certain antibacterial drugs and antitumor drugs. For example, some ototoxic drugs are chemotherapeutic drugs (such as antitumor drugs) and antibiotics. Other possible drug candidates include loop diuretics, quinine or quinine compounds, and salicylates or salicylate compounds. Therefore, disclosed herein are drugs, compositions, kits and methods for treating or preventing hearing loss caused by ototoxic drugs, wherein the ototoxic drug can be an antitumor drug (such as an ototoxic aminoglycoside antibiotic) (e.g., cisplatin), an antibiotic (such as an aminoglycoside), a loop diuretic, quinine, a quinine compound, a salicylate or a salicylate compound.

[0117] Ototoxic aminoglycoside antibiotics include but are not limited to neomycin, paromomycin, ribosomycin, viomycin, kanamycin, amikacin, tobramycin, viomycin, gentamicin, sisomicin, netilmicin, streptomycin, dibekacin, fortimycin and dihydrostreptomycin, or a combination thereof. Specific antibiotics include neomycin B, kanamycin A, kanamycin B, gentamicin C1, gentamicin C1a and gentamicin C2. Therefore, disclosed herein are drugs, compositions, kits and methods for treating or preventing drug-induced hearing loss, comprising administering a therapeutically effective amount of a pharmaceutical composition of the present disclosure to a subject who has received, is receiving or is about to receive treatment with one or more aminoglycoside drugs. In some embodiments, the ototoxic aminoglycoside antibiotic is not streptomycin. In some embodiments, the hearing impairment is not caused by an ototoxic aminoglycoside antibiotic. In some embodiments, the hearing impairment is not caused by streptomycin.

[0118] The drugs, compositions, kits, and methods disclosed herein can prevent or reduce aminoglycoside-induced hearing loss. Although aminoglycosides are particularly useful due to their rapid bactericidal effects on infections caused by aminoglycoside-susceptible bacteria, their use has so far been limited to more severe, complicated infections due to their ototoxic and nephrotoxic side effects. Therefore, aminoglycosides are considered to have a low treatment / risk ratio compared to other systemic antibiotics. Therefore, also disclosed herein are improved methods for treating infections caused by aminoglycoside-susceptible bacteria, comprising administering to a subject an antimicrobially effective amount of an aminoglycoside and a pharmaceutical composition disclosed herein. It should be recognized that recommended dosages of aminoglycosides have been established; and the methods disclosed herein are effective when aminoglycosides are administered at about 100% to about 500% (particularly about 100% to about 250%, more particularly about 100% to about 150%) of the currently recommended dosage, which are generally available on the labels and package inserts of commercially available aminoglycoside drug products. The improved methods provide for the prevention of aminoglycoside-induced hearing loss and / or tinnitus or Meniere's disease, thereby extending the therapeutic index of aminoglycoside drugs.

[0119] The disclosed pharmaceutical compositions can be administered in combination with one or more ototoxic drugs in the same dosage form. For example, an improved method is provided for treating an infection in a subject by administering an aminoglycoside antibiotic and a therapeutically effective amount of a pharmaceutical composition disclosed herein. Alternatively, the aminoglycoside antibiotic and the pharmaceutical composition disclosed herein can be administered to the subject in different dosage forms.

[0120] The one or more ototoxic drugs can also be chemotherapeutic drugs used to treat cancer in a subject. For example, an improved method is provided for treating cancer in a subject by administering a chemotherapeutic drug (such as an anti-tumor chemotherapeutic drug) and a therapeutically effective amount of a pharmaceutical composition disclosed herein.

[0121] Ototoxic anti-tumor chemotherapy drugs include cisplatin or cisplatin compounds, paclitaxel or paclitaxel compounds, and other chemotherapy drugs that are believed to cause ototoxicity-induced hearing loss (e.g., vincristine, an anti-tumor drug used to treat hematological malignancies and sarcomas). Therefore, the methods disclosed herein can be used to treat ototoxicity (e.g., drug-induced hearing loss) in subjects who have received, are receiving, or will receive anti-tumor drugs (including cisplatin or cisplatin compounds, paclitaxel or paclitaxel compounds, and other chemotherapy drugs that are believed to cause ototoxicity-induced hearing loss, such as vincristine, an anti-tumor drug used to treat hematological malignancies and sarcomas).

[0122] Central auditory dysfunction (CAPD)

[0123] Central auditory processing disorder (CAPD) involves difficulties in the central nervous system (CNS)'s ability to process auditory information. CAPD testing can include auditory discrimination testing, auditory temporal processing and pattern testing, dichotic speech testing, monaural low-redundancy speech testing, binaural interaction testing, electroacoustic testing, and electrophysiological testing.

[0124] Disclosed herein are medicaments, compositions, kits, and methods for treating or preventing hearing loss (e.g., CAPD) in a subject in need thereof, wherein the methods comprise administering a therapeutically effective amount of NMDP or a salt thereof. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reduce or eliminate CAPD. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to prevent or prevent CAPD. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reverse CAPD or at least partially restore hearing. The subject may be at risk of developing hearing loss (e.g., CAPD), or the subject may be suffering from hearing loss (e.g., CAPD).

[0125] tinnitus

[0126] Tinnitus is the perception of sound in the ears even in the absence of external auditory stimulation. Tinnitus most commonly presents as ringing in the ears; however, it can also manifest as crickets, whirring, pulsating, ocean waves, a buzzing, or even music. Tinnitus can be temporary, intermittent, or permanent; its severity ranges from a mild background ringing to an intense auditory sensation that overwhelms external sounds.

[0127] Tinnitus can be caused by one or more factors, such as the administration of or exposure to ototoxic substances (such as an aspirin overdose), exposure to brief bursts of loud noise (such as gunshots or explosions), or chronic exposure to high-decibel noise (such as airplane engine noise, high-decibel concerts, or use of high-decibel headphones), or a central auditory processing disorder as described herein.

[0128] Disclosed herein are medicaments, compositions, kits, and methods for treating or preventing hearing impairment (e.g., tinnitus) in a subject in need thereof, wherein the method comprises administering a therapeutically effective amount of NMDP or a salt thereof. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to alleviate or eliminate the hearing impairment (e.g., tinnitus). In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to prevent or prevent the hearing impairment (e.g., tinnitus) or treat its symptoms. In some embodiments, the therapeutically effective amount of NMDP or a salt thereof is an amount sufficient to reverse the hearing impairment (e.g., tinnitus) or at least partially restore hearing. The subject may be at risk of developing the hearing impairment (e.g., tinnitus), or the subject may be suffering from the hearing impairment (e.g., tinnitus).

[0129] Meniere's disease

[0130] Meniere's disease is an inner ear disorder that can cause tinnitus (tinnitus), hearing loss or decrease, vertigo, and a sensation of fullness or blockage in the ears. Vertigo attacks can occur suddenly or be followed by brief bouts of tinnitus or blurred hearing. Some people experience single episodes of vertigo separated by long intervals, while others may experience frequent episodes over several days. In some people with Meniere's disease, vertigo is so severe that it can lead to imbalance and falls. For those affected, Meniere's disease is often a serious and disabling condition.

[0131] Meniere's disease can develop at any age, but it is more common in adults between the ages of 40 and 60. The National Institute on Deafness and Other Communication Disorders (NIDCD) estimates that approximately 615,000 people in the United States are currently diagnosed with Meniere's disease, with 45,500 new cases each year. Meniere's disease typically affects only one ear. Without being bound by a particular theory, the symptoms of Meniere's disease may be caused by fluid accumulation in the inner ear cavity. Currently, there is no known cure, and improved pharmaceutical compositions and treatment methods are urgently needed.

[0132] Salts, stereoisomers, polymorphs and derivatives

[0133] Although described above with respect to specific compounds, stereoisomers, polymorphs, metabolites, derivatives and / or salts of the active compounds may also be used. Examples of therapeutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali metal or organic salts of acidic residues such as carboxylic acids. Therapeutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts formed from the parent compound, such as salts formed from non-toxic inorganic or organic acids. Conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid and isethionic acid. Therapeutically acceptable salts can be synthesized from parent compounds containing a basic or acidic moiety using conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water, in an organic solvent, or in a mixture of the two; for example, non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol or acetonitrile. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985, p. 1418).

[0134] Stereoisomers are compounds composed of the same atoms and have the same bond order, but the three-dimensional arrangement of the atoms is different and non-interchangeable. These three-dimensional structures are called configurations. Two types of stereoisomers include enantiomers and diastereomers. Enantiomers are two non-superimposable mirror-image stereoisomers, a property called chirality. "Racemate," "racemic mixture," or "racemate" refers to a mixture of equal parts of enantiomers. A "chiral center" refers to a carbon atom that is connected to four different groups. Standard techniques are used to select the appropriate chiral column, eluent, and conditions required to separate a pair of enantiomers, which are well known to those of ordinary skill in the art (e.g., see Jacques, J. et al., "Enantiomers, Racemates, and Resolutions," John Wiley and Sons, Inc. 1981). Diastereomers are two stereoisomers that are neither mirror-images nor superimposable, and their different physical properties can be easily separated by exploiting these differences. Metabolites of the above compounds arise from biochemical processes in living cells that interact with the active parent drug or other agents or compounds in vivo and include products or intermediates of any metabolic pathway.

[0135] Active Pharmaceutical Ingredients (API)

[0136] NMDP is a calcium channel blocker (CCB) in the dihydropyridine class. It is a highly lipophilic drug that rapidly crosses the blood-brain barrier. Its chemical structure and related properties are shown below. NMDP can be used to treat tinnitus associated with hearing loss, vertigo and tinnitus associated with Meniere's disease, traumatic brain injury, and subarachnoid hemorrhage. It can also prevent hearing loss during vestibular schwannoma surgery.

[0137] NMDP belongs to Class II of the Biopharmaceutics Classification System (BCS). Due to its high lipophilicity (log p = 3.41), it exhibits poor water solubility but good permeability. NMDP is a weakly basic compound with a predicted pKa value of 5.41 and is freely soluble in ethanol. NMDP has been approved by the FDA in various dosage forms, including softgels, tablets, intravenous solutions, and oral solutions (Sweetman and Martindale, 2002). Some physical properties of NMDP (NMD) are shown in the table below.

[0138] nature NMDP Molecular weight 418.4 Water-soluble 0.012mg / mL Lipid-water partition coefficient 3.05 Melting point 125℃ Number of hydrogen bond donors 1 Number of hydrogen bond acceptors 8 Dissociation constant 5.41

[0139] In humans, NMDP is rapidly absorbed after oral administration, reaching peak concentrations within 1 hour. Due to extensive first-pass metabolism, bioavailability outside the central nervous system is 100% after intravenous administration and 3-30% after oral administration. Due to high-affinity binding of NMDP to plasma proteins, only 5% of NMDP reaches the brain.

[0140] API concentration

[0141] Compositions of the present disclosure typically have a concentration equivalent to about 0.4 to 75 mg / mL NMDP, 0.5 to 40 mg / mL NMDP, 0.6 to 35 mg / mL, 0.7 to 25 mg / mL, 0.75 to 15 mg / mL NMDP, or 1 to 10 mg / mL NMDP. Suitable compositions have a concentration equivalent to at least about 0.4 mg / mL NMDP, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, or 5 mg / mL NMDP, for example, 7 mg / mL, 10 mg / mL, about 15 mg / mL, about 20, about 25, about 35, about 45, about 60, about 80, about 100, about 150, about 125, about 150, about 180, about 225, about 275, and about 400 mg / mL NMDP.

[0142] As described, the composition is delivered in dosage units, wherein administration comprises delivering one or more dosage units of about 10 to 500 μL, for example 10 to 200 μL, preferably about 50 to 150 μL. In embodiments where delivery is via the nasal mucosa, the delivery unit corresponds to the spray or jet volume provided by the device used to deliver the composition and dosage unit.

[0143] If the nasal administration volume exceeds about 200 μL, there may be a risk of the formulation being lost to the throat or through the nostrils. Therefore, in some embodiments, the nasal administration formulation should not exceed 200 μL per administration. Therefore, the volumes disclosed herein include volumes selected from 10 μL, 25 μL, 50 μL, 75 μL, 100 μL, 150 μL, 200 μL, 250 μL, 300 μL, and 350 μL, as well as 400 μL, wherein the volume can be delivered to both nostrils if desired.

[0144] In one embodiment, the composition is formulated for nasal delivery comprising a dosage unit equivalent to at least about 70 μg NMDP, for example 80, 90 or 100 μg, for example 125, 150, 200, 250 or 300 μg, for example 350, 400, 450, 500 μg, for example 550, 600, 650, 700, 750, 800, 850, 900 or 950 μg, for example 1000, 1050, 1100, 1250 or 1300 μg, for example 1350, 1400, 1450, 1500 μg, for example 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900 or 1950 μg, for example a dosage unit equivalent to 2000 μg NMDP.

[0145] Alternatively, it is defined as a composition formulated for transmucosal delivery of a dosage unit equivalent to about 70 to 2500 mg NMDP, e.g., 70 to 1800 mg, 70 to 1500 mg, 70 to 1200 mg, 70 to 1000 mg, 70 to 500 mg, 75 to 300 mg NMDP.

[0146] The present disclosure also relates to a method for administering NMDP or a pharmaceutically acceptable salt thereof to the circulatory system of an individual in need of relief from acute tinnitus or Meniere's disease or its symptoms. The therapeutic dose should be sufficient to treat acute tinnitus or Meniere's disease or its symptoms within a narrow timeframe. To produce a plasma concentration sufficient to treat acute tinnitus or Meniere's disease or its symptoms, the therapeutic dose is typically in the range of at least about 1 to about 5 mg / kg, or about 50 mg to about 250 mg. To deliver the NMDP to the circulatory system within an acceptable timeframe and without injection, the NMDP is administered to the patient's mucosa in a pharmaceutical carrier for transmucosal delivery of the NMDP.

[0147] The dosage sufficient to alleviate tinnitus or Meniere's disease or its symptoms may vary from patient to patient, and even within individual patients. For the treatment of relatively moderate acute tinnitus or Meniere's disease or its symptoms, a therapeutic dose may comprise at least 70 μg NMDP, preferably at least 100 μg NMDP, at least 150 μg NMDP, for example, 200 μg NMDP. For the treatment of more severe acute tinnitus or Meniere's disease or its symptoms, a therapeutic dose comprises at least 250 μg NMDP, preferably at least 300 μg NMDP, at least 400 μg NMDP, for example, 500 μg NMDP, 1000 μg NMDP, 1500 μg NMDP, 2000 μg NMDP. In cases where a patient suffers from severe acute tinnitus or Meniere's disease or its symptoms, or where the patient has developed tolerance to NMDP, a higher dose may be required and administered in accordance with the present disclosure. Such high doses include therapeutic doses comprising 25 mg NMDP, at least 30 mg NMDP, for example, at least 40 mg NMDP, at least 50 mg NMDP, at least 100 mg, at least 150 mg, at least 200, at least 400 mg, at least 600 mg, at least 1000 mg. Even higher doses may be required, for example, therapeutic doses of 1300 mg NMDP, preferably at least 1400 mg NMDP, at least 1500 mg NMDP, for example 1700 mg NMDP. In some embodiments, treatment may involve patients requiring a therapeutic dose comprising 1800 to 2500 mg NMDP.

[0148] As described, the concentrations of the transmucosal delivery compositions of the present disclosure are more effective than compositions known to those skilled in the art. In one embodiment, the composition is formulated so that the therapeutic dose comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 25, 30, 35, 40, 45, 50, 100, or 300 dosage units.

[0149] An important aspect of the present disclosure is that relief of tinnitus or Meniere's disease or its symptoms can be achieved very quickly after administration of NMDP. Thus, relief of acute tinnitus or Meniere's disease or its symptoms should be achieved shortly after the first delivery of a dosage unit or therapeutic dose, such that the onset time of administration of the composition is less than 10 minutes, e.g., less than 9 minutes or less than 8 minutes. In some embodiments, the onset time of the composition can be greater than 10 minutes, e.g., 15 minutes, 20 minutes, 25 minutes, 30 minutes, or greater than 30 minutes.

[0150] In some embodiments, in addition to a very short onset of action, the relief of tinnitus or Meniere's disease or its symptoms lasts for at least 30 minutes. In some embodiments, the duration of action of the composition administered lasts for a period of at least 30 minutes, at least 60 minutes, at least 90 minutes, at least 120 minutes, at least 240 minutes, at least 360 minutes, at least 480 minutes, at least 600 minutes, at least 720 minutes, at least 840 minutes, at least 960 minutes, or at least 1080 minutes, 20 hours, 22 hours, 24 hours, 36 hours, 48 hours, or 60 hours.

[0151] Compared to intravenous administration, which has a very rapid onset but a very short duration of action, the composition exhibits a pseudo "sustained release" effect. Following administration, the intranasal compositions described herein typically have a bioavailability of not less than 75% of that of intravenous administration, for example, not less than 80% or not less than 90% of that of intravenous administration. Bioavailability can be determined by AUC, which is known to those skilled in the art.

[0152] The methods of the present disclosure comprise administering dosage units comprising about 70 to 2500 mg of NMDP, said administration resulting in an intranasal Cmax / intravenous Cmax (Cmax) within the therapeutic dose range of about 70 to 2500 mg, as the dosage units delivering an equivalent amount of NMDP increase. max,nasal / C max,iv ) ratio decreased.

[0153] API formulations

[0154] To facilitate intranasal administration of NMDP, an effective amount of NMDP should be dissolved or finely dispersed in a small volume of liquid carrier, for example, less than about 1,000 μL (microliters), preferably less than 300 μL, for example, less than 150 μL. Larger volumes will flow forward through the nostrils or backward toward the pharynx, where excess liquid is swallowed. Therefore, if the administration volume is too large, some NMDP may be lost from the absorption site, and even if it is possible, repeated administration of the correct dose of the therapeutic agent will be difficult. Therefore, it is ideal to have a high dissolved NMDP concentration or dispersed NMDP content in the small volume of liquid carrier for intranasal administration. In addition, to support drug absorption through the nasal mucosa, two natural protective functions must be bypassed: mucociliary clearance (MCC) and the barrier properties of the tissue. Ideally, an effective dose can bypass the natural properties of the mucus layer as a protective layer (i.e., to increase drug absorption) and MCC as an effective cleaning mechanism. In other words, it is ideal to increase the residence time of the administered dose on the mucus layer. Surfactants suitable for the present invention are nonionic surfactants. Suitable nonionic surfactants for use in the present invention are polysorbates, particularly polysorbate 20 (Tween 20) and polysorbate 80 (Tween 80). Tween 80 (polyoxyethylene sorbitan monooleate) is an oleate, and Tween 20 (polyoxyethylene sorbitan monolaurate) is a laurate. Both are listed on the U.S. Food and Drug Administration (FDA) list of inactive ingredients for nasal sprays. Both Tween 20 and Tween 80 are miscible with water. The chemical structure of Tween 80 is shown below.

[0155]

[0156] Water-soluble cellulosic polymers suitable for use in the present invention include, but are not limited to, hydroxypropyl methylcellulose (HPMC; also known as hypromellose), hydroxyethylcellulose (HEC), carboxymethylcellulose (CMC), hydroxypropylcellulose (HPC), sodium carboxymethylcellulose, sodium carboxymethylcellulose with microcrystalline cellulose (MCC), xanthan gum, guar gum, gum arabic, natural gums (such as tragacanth), and / or sodium carboxymethylcellulose. These polymers are also listed on the US Food and Drug Administration (FDA) list of inactive ingredients for nasal sprays. HPMC is a preferred polymer for use in the present invention. It is a nonionic, water-soluble cellulose derivative with hydrogen bonding potential and stabilizing properties, preventing crystallization of amorphous materials. HPMC at a concentration of approximately 1% in phosphate buffer (pH 6.8) can also increase drug solubility. In some embodiments, the viscosity of a 2% (w / v) aqueous solution of HPMC used in the formulation at 20°C ranges from approximately 2–4,000 mPa.s, approximately 4–60 mPa.s, and / or approximately 4–6 mPa.s. In certain embodiments, the HPMC is E6, E50, or E4M. In some embodiments, the molecular weight of the preferred HPMC for use in the formulation may be about 50 to about 100 kDa, about 50 to about 150 kDa, about 50 to about 200 kDa, about 50 to about 250 kDa, about 50 to about 300 kDa, about 50 to about 350 kDa, about 50 to about 400 kDa, about 50 to about 500 kDa, about 50 to about 600 kDa, about 50 to about 700 kDa, about 50 to about 800 kDa, about 50 to about 900 kDa, about 50 to about 1100 kDa, about 50 to about 1200 kDa, about 50 to about 1300 kDa, about 50 to about 1400 kDa, about 50 to about 1500 kDa, about 50 to about 1600 kDa, about 50 to about 1700 kDa, about 50 to about 1800 kDa, about 50 to about 1900 kDa, about 50 to about 2000 kDa, about 50 to about 2500 kDa, about 50 to about 3000 kDa, about 50 to about 350 kDa, about 50 to about 400 kDa, about 50 to about 500 kDa, about 50 to about 600 kDa, about 50 to about 700 kDa, about 50 to about 800 kDa, about 50 to about 900 kDa, about 50 to about 1100 kDa, about In some embodiments, the NMDP of the present invention can be a water-soluble NMDP having a molecular weight of about 50 to about 1500kDa, about 100 to about 150kDa, about 100 to about 200kDa, about 100 to about 250kDa, about 100 to about 300kDa, about 100 to about 350kDa, about 100 to about 400kDa, about 100 to about 500kDa, about 100 to about 600kDa, about 100 to about 700kDa, about 100 to about 800kDa, about 100 to about 900kDa, about 100 to about 1100kDa or about 100 to about 1500kDa. In some embodiments, polyethylene glycol (PEG) or its methoxy derivative (such as methoxypolyethylene glycol (mPEG)), or the combination of mPEG / PEG, can be used as a good solvent for the NMDP of poor water solubility. In some embodiments, a mixture of low molecular weight PEG, mPEG and / or PEG and mPEG can be used. In some embodiments, PEG, mPEG, and / or mixtures of PEG and mPEG can be used to prepare low viscosity formulations.

[0157] The structure of polyethylene glycol (PEG) is shown in I:

[0158] I: HO-(CH2CH2O) n-H, wherein n is a number in the range of 1 to 25.

[0159] The structure of alkoxy polyethylene glycol is shown in II:

[0160] II: RO-(CH2CH2O) n -H, wherein R is methyl, ethyl, n-propyl, isopropyl or cyclopropyl; and n is the average number of oxyethylene repeating units, which ranges from about 1 to about 25.

[0161] Determination of the effect of pharmaceutical compositions

[0162] The composition is intended to treat, alleviate or reduce acute or breakthrough tinnitus, Meniere's disease or tinnitus / Meniere's disease symptoms based on the following assessments: a) measurement of changes in auditory brainstem response (ABR) thresholds; b) changes in auditory speech recognition as measured by a speech-in-noise test; c) changes in auditory speech recognition as measured by a digit-in-noise test; d) changes in low-frequency hearing thresholds; e) changes in the incidence of adverse events after administration of the pharmaceutical composition; f) changes in the severity of tinnitus or Meniere's disease; g) changes in the loudness of tinnitus or Meniere's disease; h) changes in the severity of vertigo; i) changes in the sensation of fullness in the ears; j) changes in dizziness; and k) changes in hair cell function observed by changes in ABR thresholds after administration of the pharmaceutical composition.

[0163] On the other hand, compositions of the present disclosure, dosage unit, purposes and method are characterised in that the treatment of measuring as described herein is to the effect of acute tinnitus, Meniere's disease or its symptom.According to a method of the disclosure recording tinnitus, Meniere's disease or its symptom, comprise measuring the onset time of tinnitus / Meniere's disease or its symptom relief.For example, timing (such as starting stopwatch) before starting treatment, when experimenter determines to feel significant tinnitus / Meniere's disease or its symptom relief, record time (such as stopping stopwatch).Compositions of the present disclosure, after administration, is measured by PID, and after delivering no more than two dosage units (preferably one dosage unit), the relief score scope of tinnitus / Meniere's disease or its symptom is 2 to 7 (such as 2,3,4,5,6,7, preferably 3,4,5,6).

[0164] Success is considered if at least 50% of subjects experience an onset of effect within 15 minutes of treatment. Similarly, duration of effect can be measured as the difference between the onset of effect and the point at which the subject declares the effect has ceased, or the time the subject takes rescue medication, whichever occurs first. Success is considered successful if at least 50% of subjects experience relief from tinnitus / Meniere's disease or its symptoms for at least half an hour.

[0165] Another measure was the intensity of tinnitus / Meniere's disease or its symptoms (TI), using an 11-point numeric rating scale (0 = no tinnitus / symptom, 10 = unbearable tinnitus / symptom). i Time point Ti The intensity of tinnitus / symptoms was measured at one or more time points (T i A 40% reduction in the mean TI within 15 minutes of treatment was considered successful, although other time points and intervals could be used.

[0166] TI0 is the baseline tinnitus / symptom intensity (scored according to the above scale) before treatment (time T0), and the tinnitus / symptom intensity difference (TID) is the difference between TI0 and each time point after treatment (TI i A mean TID of 2 within 15 minutes of treatment was considered successful.

[0167] Further measurements were the area under the TID curve or the sum of tinnitus / symptom intensity differences (STID), with TI measured at the above time points. A 4-hour average STID of 3 was considered successful.

[0168] A method involving the tinnitus / Meniere's disease or symptom intensity scale described herein, wherein tinnitus / symptom relief is measured by the tinnitus / symptom intensity difference (TID), which is based on the TI0 score measured near the time of administration and the time TI0 after administration. i The measured score has a difference of at least 30% (e.g., at least 40%). The time after administration can be selected from 3 minutes, 5 minutes, 7 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, etc. after administration to evaluate the immediate effect of administration. If it is necessary to measure the duration of treatment, tinnitus / Meniere's disease or its symptom relief is based on the TI0 score measured immediately before administration and the score at time points such as 45 minutes, 60 minutes, 75 minutes, 90 minutes, 120 minutes after administration to calculate the TID, or the desired time period can be selected separately.

[0169] The score for relief of tinnitus / Meniere's disease or its symptoms can be measured as described herein, or using a scale of 1-100% (100% being intolerable tinnitus / symptoms described by the patient and 0% being completely free of tinnitus / symptoms), preferably with a score of at least 30% from the onset to the maximum relief effect achieved.

[0170] Another measurement is the sum of the tinnitus / symptom intensity differences (STID) calculated based on the PIO score measured immediately before administration and the PII score at any time point after administration, and the time after administration includes the time described herein. In one embodiment, STID is calculated based on at least 2 values measured within 30 minutes (preferably 45 minutes, more preferably 60 minutes, such as 90 minutes), and can also be measured based on at least 5 values (such as 7, preferably 10, such as 11, 12 or 13).

[0171] In other embodiments, a therapeutic dose of NMDP or a salt thereof can also be administered to one or more mucosa selected from the oral mucosa, respiratory mucosa (e.g., tracheal mucosa, and / or pulmonary mucosa). Furthermore, the therapeutic dose disclosed herein can be administered to multiple sites in the same treatment, or the patient can select the route of administration based on individual circumstances. If intranasal administration causes nasal irritation, acute nocturnal tinnitus / Meniere's disease or its symptoms can be treated by oral administration.

[0172] Area under the curve

[0173] As used herein, the peak concentration of NMDP in plasma (C max )”, “Area under the plasma NMDP concentration-time curve (AUC)”, “Time for plasma NMDP to reach maximum concentration (t max )” is a pharmacokinetic parameter known to those skilled in the art [Laursen et al., Eur. J. Endocrinol. 135:309-315, 1996]. The “concentration-time curve” is used to measure the change in the concentration of NMDP in the serum of a subject over time after administration of a dose of NMDP via the intranasal or mucosal route. “C max " refers to the maximum concentration of NMDP in the ear structure or serum after a single administration of NMDP to the subject. "t max ” refers to the time it takes for NMDP to reach its maximum concentration in serum or otic structures after a single administration of NMDP to a subject.

[0174] As used herein, the area under the NMDP concentration-time curve (AUC) in plasma or otic structures is calculated according to the linear trapezoidal rule and the residual area is added. A 23% decrease or a 30% increase between two doses can be detected with a probability of 90% (type II error β = 10%). The "delivery rate" or "absorption rate" is determined by comparing the maximum concentration (C max ) time (t max ) to make an estimate. C max and t max All analyses were performed using nonparametric methods. The pharmacokinetics of NMDP administered intramuscularly, subcutaneously, intravenously, and intranasally were compared using analysis of variance (ANOVA). Pairwise comparisons were assessed for significance using the Bonferroni-Holmes sequential method. Dose-response relationships between the three intranasal doses were estimated using regression analysis, with P < 0.05 considered significant. Results are presented as mean ± standard error (SEM).

[0175] In one embodiment, the instantaneous serum level of NMDP administered intranasally reaches about 5 pg / mL to about 280 pg / mL, about 10 pg / mL to about 100 pg / mL, about 20 pg / mL to about 50 pg / mL, or 25 pg / mL to about 250 pg / mL. In intranasal formulations, the instantaneous serum level of NMDP achieved may be lower or higher than the level typically achieved with other routes of administration. In some embodiments, the beneficial effects of intranasal administration are similar to those achieved when the steady-state serum NMDP level is about 5 ng / dL to about 180 ng / dL.

[0176] Comparing the total concentration-time curve area (AUC) or mean concentration of NMDP in subjects who were administered NMDP intranasally with those who were administered NMDP by other routes (such as oral administration) provides a basis for determining the bioequivalence of different administration routes. Although the administration routes or concentration-time curves are different, if the AUC or mean concentrations are similar, the biological effects generally produced are also similar. Therefore, in one embodiment, the present disclosure contemplates administering the composition intranasally to achieve an average serum NMDP concentration of about 5 pg / mL to about 250 pg / mL over 24 hours.

[0177] In one embodiment, NMDP is administered intranasally to achieve a transient concentration level in the ear structures of about 5 pg / mL to about 280 pg / mL, about 10 pg / mL to about 100 pg / mL, about 20 pg / mL to about 50 pg / mL, or 25 pg / mL to about 250 pg / mL. In the intranasal formulation, the transient ear serum levels of NMDP achieved may be lower or higher than those typically achieved with other routes of administration.

[0178] The peak plasma concentration achieved by intravenous administration of NMDP is associated with side effects such as respiratory depression. In some embodiments, the peak plasma concentration of the pharmaceutical composition is sufficient to produce the desired effect (and the onset of action is rapid and the duration is sufficient). Therefore, the present disclosure also relates to a composition, wherein within a therapeutic dose range of about 20 to 2500 mg, the peak plasma concentration after administration of no more than two dosage units is not less than 5% and not more than 75% of the peak plasma concentration obtained by intravenous administration of the dosage unit, preferably not less than 30% and not more than 75%.

[0179] In some embodiments, repeated administration of a dosage unit does not result in an increase in peak plasma concentration. When administered intravenously, repeated administration can continuously increase plasma concentrations to undesirably high levels. In some embodiments, repeated transmucosal administration of a dosage unit of NMDP or a salt thereof does not continuously increase plasma concentrations. In some embodiments, the method is such that, within a therapeutic dose range of about 50 to 2500 mg, when comparing equivalent amounts of NMDP delivered by two administration routes (intranasal vs. intravenous), C max,nasal / C max,ivThe ratio decreases as the dosage unit increases.

[0180] preparation

[0181] The compound or its therapeutically acceptable salt can be formulated into a pharmaceutical composition. Such compositions can be administered by inhalation spray, intranasal administration, and other dosage forms, wherein the dosage form comprises a conventional non-toxic therapeutically acceptable carrier, adjuvant, and excipient (if necessary). In some embodiments, the pharmaceutical composition can be administered by oral dosage form (such as tablets).

[0182] Pharmaceutical formulations are discussed in, for example, Hoover, John E., Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa., 1975), and Liberman, HA and Lachman, L., eds., Pharmaceutical Dosage Forms (Marcel Decker, New York, NY, 1980).

[0183] The active compound (or its therapeutically acceptable salt) can be administered alone or in the form of a pharmaceutical composition in which the active compound is mixed with one or more therapeutically acceptable carriers, excipients, or diluents. Pharmaceutical compositions can be formulated using conventional methods using one or more therapeutically acceptable carriers containing excipients and adjuvants that aid in processing the active compound into a therapeutically useful formulation. Suitable formulations depend on the chosen route of administration.

[0184] Additionally, the formulations may contain conventional carriers, such as plasticizers, pigments, colorants, glidants, stabilizers, pore-forming agents, and surfactants.

[0185] Suitable diluents include, but are not limited to, dibasic calcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starch, pregelatinized starch, silicon dioxide, titanium dioxide, magnesium aluminum silicate, and powdered sugar. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic gums (such as gum arabic, tragacanth, and sodium alginate), cellulose (including hydroxypropyl methylcellulose, hydroxypropyl cellulose, ethyl cellulose, and wigamy gum), and synthetic polymers (such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylic acid copolymers, polyacrylic acid / polymethacrylic acid, and polyvinyl pyrrolidone). Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, polyethylene glycol, talc, and mineral oil. Disintegrants may include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, pregelatinized starch, clay, cellulose, alginates, gums, or cross-linked polymers such as cross-linked PVP (cross-linked polyvinyl pyrrolidone from GAF Chemical Corp.) Stabilizers are used to inhibit or delay drug decomposition reactions, such as oxidation reactions.

[0186] Surfactants can be anionic, cationic, amphoteric or nonionic surfactants. Suitable anionic surfactants include, but are not limited to, surfactants containing carboxylate, sulfonate and sulfate ions. Examples of anionic surfactants include sodium, potassium, and ammonium salts of long-chain alkyl sulfonates and alkylaryl sulfonates (such as sodium dodecylbenzenesulfonate); sodium dialkyl sulfosuccinates (such as sodium bis-(2-ethylthiohexyl)sulfosuccinate); and alkyl sulfates (such as sodium lauryl sulfate). Cationic surfactants include, but are not limited to, quaternary ammonium compounds (such as benzalkonium chloride, benzethonium chloride, cetyltrimethylammonium bromide, stearyldimethylbenzyl ammonium chloride, polyoxyethylene, and coconut amine). Examples of nonionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG-1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, 401, stearyl monoisopropanolamide, and polyoxyethylene hydrogenated tallow amide. Examples of amphoteric surfactants include sodium N-dodecyl-β-alanine, sodium N-lauryl-β-iminodipropionate, myristamidoamphoacetate, lauryl betaine, and lauryl sulfobetaine.

[0187] If desired, tablets, beads, granules, or microparticles used for administration of the pharmaceutical composition may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, dyes, pH buffering agents, or preservatives.

[0188] The pharmaceutical compound can be complexed with other agents as part of a therapeutic formulation. Pharmaceutical compositions can take the form of, for example, liquid formulations. Liquid formulations for intranasal administration are prepared in water or other aqueous carriers and may include various suspending agents such as methylcellulose, alginates, tragacanth, pectin, kelp gum, carrageenan, gum arabic, polyvinyl pyrrolidone, and polyvinyl alcohol. Liquid formulations can also include solutions, emulsions, syrups, and elixirs containing the active compound along with a wetting agent, sweetener, and coloring and flavoring agents. Various liquid and powder formulations can be prepared by conventional methods for inhalation by the patient.

[0189] Additionally, immediate release compositions may be formulated in admixture with delayed / sustained release compositions.

[0190] In some embodiments, NMDP is formulated as the sole active pharmaceutical ingredient (API) in a dosage form. Such NMDP dosage forms can be used alone or in combination with one or more additional dosage forms comprising one or more active pharmaceutical ingredients to prevent or treat hearing loss. In this case, the daily dose of NMDP can be conveniently provided in a single dosage form as described herein, or can be divided into two, three, four, or more doses.

[0191] As described above, the composition comprising NMDP or its salt is administered transmucosally by contacting the composition in a suitable dosage form with the mucosal tissue of the vagina, nose, rectum or mouth. In one embodiment, the composition is administered via the nasal mucosa, i.e., intranasally. The nasal mucosa provides a useful anatomical site for systemic delivery. The nasal tissue is rich in blood vessels, providing an ideal site for rapid and efficient absorption. The adult nasal cavity has a capacity of approximately 20 mL, and due to the presence of microvilli on the pseudostratified columnar epithelial cells of the nasal mucosa, the surface area for drug absorption is approximately 180 cm 2 .

[0192] The nasal preparation comprising the above composition can take a variety of forms, such as nasal drops, nasal sprays, gels, ointments, creams, powders or suspensions, and can be administered using a dispenser or other device as needed. Various dispensers and delivery vehicles are known in the art, including single-dose ampoules, atomizers, sprayers, pumps, nasal cotton pads, nasal sponges, nasal capsules, and the like.

[0193] More generally, the formulation can take solid, semisolid or liquid form. For solid forms, the components can be mixed together by mixing, drum mixing, freeze drying, solvent evaporation, co-grinding, spray drying and other techniques known in the art. Such solid-state formulations preferably provide dry powdered compositions having a particle size in the range of about 5 to about 500 microns, more preferably 50 to 250 microns, for intranasal administration.

[0194] Semisolid formulations suitable for intranasal administration can take the form of aqueous or oily gels or ointments. For example, the above components can be mixed with microspheres of starch, gelatin, collagen, dextran, polylactic acid, polyglycolic acid, or other similar materials capable of forming hydrophilic gels. The microspheres can be loaded with the drug and, after administration, form a gel that adheres to the nasal mucosa.

[0195] Nasal spray preparations

[0196] In one embodiment, the nasal formulation is in liquid form, which may include an aqueous solution, aqueous suspension, oil solution, oil suspension, or emulsion, depending on the physicochemical properties of the composition components. The liquid formulation is administered by nasal spray or nasal drops, using devices known in the art, including a nebulizer capable of delivering a selected volume of the formulation in the form of a droplet aerosol. For example, commercially available spray pumps with a delivery volume of 50 or 100 μL, such as those available from Valois (Congers, NY), are available in adult and child-sized spray heads. In one embodiment, the composition comprising at least NMDP is administered nasally by aerosol spray at a daily volume of about 30 to about 200 μL.

[0197] Liquid preparations can be prepared by known processes. For example, aqueous nasal preparations can be prepared by dissolving, suspending or emulsifying NMDP or its salt in water, buffer or other aqueous media, or emulsifying it in an oily base (such as pharmaceutically acceptable oils such as olive oil, lanolin, silicone oil, glycerol, fatty acids, etc.).

[0198] Compositions of the present disclosure can be applied to the nasal cavity by liquid, spray, aerosol, atomizer or semisolid preparation.Semisolid preparation can be based on gel, water-in-oil or oil-in-water cream, or hydrophilic / lipophilic ointment.Compositions can include the fine particles / crystals of the activating agent or activating agent of molecular dispersion (dissolution, solubilization, etc.).Described composition can be applied by nasal sprayer, metered sprayer, squeeze bottle, liquid dropper, disposable single-dose dropper, atomizer, cartridge system, single-dose pump, double-dose pump, multi-dose pump or any other device with single-dose ampoule.For example, compositions of the present disclosure can be stored in or delivered by the spray or aerosol device / container described in detail in " Remington's Pharmaceutical Science " (16th edition, Chapter 83 and Chapter 92).

[0199] Regarding the spray device, it should be noted that single (unit) dose or multi-dose systems can be used. Typically, the spray device includes a bottle and a pump; such devices are commercially available from various sources. Typically, the volume of liquid released by a single spray actuation is 5 to 250 μL per nostril per single application, and the concentration of the active ingredient in the formulation can be easily adjusted so that one or more nostril sprays can meet the dosing regimen. The present disclosure also provides a spray device or dosage chamber for use in an intranasal delivery device loaded with the above-mentioned composition.

[0200] As used herein, intranasal administration or intranasal delivery includes administering the composition into the nostrils of a mammal to the mucous membranes of the nasal passages or nasal cavity. Such formulations can be administered, for example, by nasal sprays, nasal inhalers, nasal drops, aerosols, propellants, pressure dispersers, aqueous aerosols, atomizers, nasal suspensions, drops, nasal gels, nasal powders, nasal ointments, and nasal creams, using any modern or conventional device. Administration of the disclosed compositions can also be performed using nasal tampons or nasal sponges containing the compositions.

[0201] In some embodiments, a therapeutically effective amount of NMDP is administered in a composition comprising the following ingredients: a carrier, a citrate buffer, and benzalkonium chloride. In some embodiments, the composition may further comprise a non-aqueous solvent. In some embodiments, the composition also comprises a surfactant. In some embodiments, the carrier may be present in an amount of about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90% or 95% (w / w). In some embodiments, the amount of the non-aqueous solvent may be in the range of about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 95% (w / w). In some embodiments, the amount of surfactant in the composition may range from about 0.00001%, about 0.0001%, about 0.001%, about 0.01%, about 0.1%, about 1%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, or about 60%.

[0202] In some embodiments, a therapeutically effective amount of an NMDP is administered in an intranasal composition that does not contain a nonionic surfactant.

[0203] In some embodiments, the intranasal composition without nonionic surfactant comprises PEG, mPEG, water and NMDP. In some embodiments, the intranasal composition without nonionic surfactant comprises a composition of the following ingredients: about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90% or about 80% to about 90% (w / w) mPEG or mPEG350, about 10% to about 50%, about 20% to about 50%, about 30% to about 50% or about 40% to about 50% (w / w) PEG or PEG400, and about 5% to about 20%, about 10% to about 20% or about 15% to about 20% (w / w) water and NMDP. In some embodiments, the concentration of NMDP in the intranasal composition without nonionic surfactant is at least 10 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / mL, at least 30 mg / mL, at least 35 mg / mL, at least 40 mg / mL, at least 45 mg / mL, at least 50 mg / mL, at least 55 mg / mL, at least 60 mg / mL, at least 65 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 95 mg / mL, at least 100 mg / mL, at least 110 mg / mL or greater than 110 mg / mL. In some embodiments, a therapeutically effective amount of NMDP is administered in an intranasal composition comprising about 70% (w / w) mPEG350, about 20% (w / w) PEG400 and about 10% (w / w) water. In some embodiments, the intranasal composition has an NMDP concentration of at least 10 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / mL, at least 30 mg / mL, at least 35 mg / mL, at least 40 mg / mL, at least 45 mg / mL, at least 50 mg / mL, at least 55 mg / mL, at least 60 mg / mL, at least 65 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 95 mg / mL, at least 100 mg / mL, at least 110 mg / mL, or greater than 110 mg / mL.

[0204] In some embodiments, the intranasal composition without a nonionic surfactant comprises PEG, mPEG, water, ethanol, and NMDP. In some embodiments, the intranasal composition without a nonionic surfactant comprises a composition of about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, or about 80% to about 90% (w / w) mPEG or mPEG350, about 10% to about 50%, about 20% to about 50%, about 30% to about 50% Or about 40% to about 50% (w / w) PEG or PEG400, about 5% to about 20%, about 10% to about 20% or about 15% to about 20% (w / w) water, about 1% to about 10%, about 2% to about 10%, about 3% to about 10%, about 4% to about 10%, about 5% to about 10%, about 6% to about 10%, about 7% to about 10%, about 8% to about 10% or about 9% to about 10% ethanol, and NMDP. In some embodiments, the concentration of NMDP in the intranasal composition is at least 10 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / mL, at least 30 mg / mL, at least 35 mg / mL, at least 40 mg / mL, at least 45 mg / mL, at least 50 mg / mL, at least 55 mg / mL, at least 60 mg / mL, at least 65 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 95 mg / mL, at least 100 mg / mL, at least 110 mg / mL, or greater than 110 mg / mL.

[0205] In some embodiments, a therapeutically effective amount of NMDP is administered in a composition comprising one or more forms of alkoxypolyethylene glycol.

[0206] RO-(CH2CH2O) n -H, wherein R is methyl, ethyl, n-propyl, isopropyl or cyclopropyl. In some embodiments, the average number of oxyethylene repeating units is a value in the range of about 1 to about 25. Alkoxy polyethylene glycol represented by formula I:

[0207] RO-(CH2CH2O) n -H(I), wherein R is (C1-C6)alkyl; n is the average number of ethylene oxide repeating units, and has a value ranging from about 1 to about 25. The term "(C1-C6)alkyl" refers to an alkyl group having 1 to 6 carbon atoms. Representative alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopropylmethyl, cyclopentyl, cyclobutylmethyl, cyclobutylethyl, cyclohexyl, cyclopropylpropyl, cyclobutylethyl, and cyclopentylmethyl.

[0208] In some embodiments, ethylene glycol may be used alone or as a mixture of two or more methoxy-n-ethylene glycols.

[0209] In certain embodiments, the alkoxy polyethylene glycol is polyethylene glycol 200 (PEG 200), polyethylene glycol 400 (PEG 400), polyethylene glycol 600 (PEG 600), polyethylene glycol 1000 (PEG 1000). The term "PEG 200" refers to polyethylene glycol with an average molecular weight of about 200 daltons; "PEG 400" refers to polyethylene glycol with an average molecular weight of about 400 daltons; "PEG 600" refers to polyethylene glycol with an average molecular weight of about 600 daltons; and "PEG 1000" refers to polyethylene glycol with an average molecular weight of about 1000 daltons.

[0210] In certain embodiments, the alkoxy polyethylene glycol is methoxy polyethylene glycol 350 (mPEG 350), methoxy polyethylene glycol 550 (mPEG 550), or methoxy polyethylene glycol 750 (mPEG 750). The term "mPEG 350" refers to a methoxy polyethylene glycol having an average molecular weight of about 350, and in certain embodiments, the average value of n in Formula I is 7.2; "mPEG 550" refers to a methoxy polyethylene glycol having an average molecular weight of about 550, and the average value of n is 11.8; "mPEG 750" refers to a methoxy polyethylene glycol having an average molecular weight of about 750, and the average value of n is 16.3.

[0211] Particularly preferred alkoxy polyethylene glycols include Carbowax, commercially available from The Dow Chemical Company. TM mPEG 350, Carbowax TM mPEG 550 or Carbowax TM mPEG 750. mPEG 350 and mPEG 550 are colorless liquids that are miscible in all proportions with water, alcohols (such as methanol, ethanol, n-propanol, glycerol), and various oils, with a boiling point of approximately 155°C. It should be understood that alkoxy polyethylene glycols exist under other names, such as methoxy polyethylene glycol, also known as monomethyl polyethylene glycol and polyethylene glycol methyl ether.

[0212] In some embodiments, the composition can be optimized for bioadhesion, mucoadhesion, viscosity, and nebulization suitability. For example, mPEG 350 at the same concentration as PEG 200 can still dissolve the therapeutic agent, but the resulting composition has a lower viscosity. In certain embodiments, the lower viscosity compared to low molecular weight PEG 200 has a positive impact on nebulization suitability, which is particularly important in formulations that require nebulization.

[0213] In some embodiments, the composition comprises a component selected from polyethylene glycol, propylene glycol, methoxypropylene glycol, ethanol, and water. In some embodiments, NMDP is dissolved in one or more forms of alkoxy polyethylene glycol to reduce viscosity, and the resulting low-viscosity formulation can be prepared as a nasal spray formulation. At 20°C, the viscosity of the resulting pharmaceutical composition ranges from about 1.5 cP to about 60 cP, about 2 cP to about 50 cP, about 3 cP to about 40 cP, about 4 cP to about 30 cP, or about 5 cP to about 25 cP. In specific embodiments, the alkoxy polyethylene glycol comprises from about 0.1% (w / w) to about 80% (w / w), about 0.5% (w / w) to about 70% (w / w) of the composition; in other specific embodiments, it comprises from about 5% (w / w) to about 80% (w / w), about 30% (w / w) to about 75% (w / w), or about 40% (w / w) to about 70% (w / w). For a particular hydrophilic drug, the alkoxypolyethylene glycol may comprise from about 0.1% (w / w) to about 80% (w / w), from about 0.5% (w / w) to about 70% (w / w), or from about 1% (w / w) to about 60% of the composition; for a particular lipophilic drug, it may comprise from about 1% (w / w) to about 80% (w / w), from about 2% (w / w) to about 65% (w / w), or from about 5% (w / w) to about 50%. In addition, the therapeutic agent may comprise from about 0.001% (w / v) to about 20% (w / v) or from about 0.1% (w / v) to about 10% (w / v) of the composition.

[0214] The pH range of the pharmaceutical composition can be about 4.5 to about 8.5, about 4.5 to about 7.5, about 4.5 to about 6.5, about 5.5 to about 8.5, about 6.5 to about 8.5, or about 5.5 to about 7.5.

[0215] By using one or more of the alkoxy polyethylene glycols described herein, the resulting pharmaceutical compositions can be optimized for bioadhesion, viscosity, and nebulization suitability. For example, mPEG 350 at concentrations equivalent to PEG 200 can still dissolve the therapeutic agent, but the resulting composition has a lower viscosity. Compared to low molecular weight PEG 200, this substitution has an unexpected positive impact on nebulization suitability, which is critical in formulations that require nebulization.

[0216] It should be understood that the formulation may contain excipients required for formulation, stability and / or bioavailability, exemplary excipients include sugars (glucose, sorbitol, mannitol, sucrose), absorption enhancers (chitosan), thickeners and stability enhancers (cellulose, polyvinyl pyrrolidone, starch, etc.), buffers, preservatives and / or acids and bases for adjusting pH.

[0217] In one embodiment, an absorption enhancing component is included, exemplary absorption enhancing components include surfactant acids (such as cholic acid, glycocholic acid, taurocholic acid and other cholic acid derivatives), chitosan and cyclodextrin. In some embodiments, the formulation includes cyclodextrin.

[0218] Exemplary surfactants include nonylphenol polyoxyethylene ether, octylphenol polyoxyethylene ether, Tween, Span, sodium lauryl sulfate and sorbitan monopalmitate; exemplary absorption enhancers include bile salts and derivatives thereof, fusidic acid and derivatives thereof, oleic acid, lecithin, lysolecithin, dodecylphosphatidylcholine (DDPC), sucrose monolaurate, n-dodecyl-β-D-maltopyranoside, pectin, chitosan, α-, β- and γ-cyclodextrin and derivatives thereof, polyethylene glycol caprylic acid / capric acid glyceride and derivatives thereof (such as Softigen and Labrasol); exemplary water-absorbing polymers include polyethylene glycol with an average molecular weight of 200 to 7500, propylene glycol or mixtures thereof, or single glycols (such as tetraethylene glycol and pentaethylene glycol); exemplary alcohols include ethanol, isopropyl alcohol; exemplary lipids include vegetable oils, soybean oil, Peanut oil, coconut oil, corn oil, olive oil, sunflower oil, monoglycerides, diglycerides, mono / diglycerides, mono / di / triglycerides; exemplary osmotic pressure control agents include glycerol, glucose, maltose, sucrose, mannitol, xylitol, various salts (such as sodium chloride); exemplary pH control agents include buffers, acids (such as nitric acid, phosphoric acid or acetic acid); exemplary preservatives include methyl paraben, phenylethanol or benzoic acid; exemplary propellants include butane or air displacers (such as nitrogen); excipients for adjusting the hydrophilic-lipophilic balance (HLB) of the formulation include Tween 20, 25, 40, 45, 65, 85, Span 20-80, polyoxyethylene lauryl ether (Brij) 30-98, gum arabic; exemplary enzyme inhibitors include aprotinin and other peptidase inhibitors, diisopropyl fluorophosphate (DFP), carbomer; exemplary stabilizers include cyclodextrin.

[0219] It may be beneficial to include additional compounds that enhance the solubility of the therapeutic agent. Examples of such solubilizers include alcohols and polyols (e.g., ethanol, isopropanol, butyl alcohol, benzyl alcohol, ethylene glycol, propylene glycol, butylene glycol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, diethylene glycol monoethyl ether, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyethylene glycol mono / dicaprylic / capric glycerides, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclopentane, hydroxypropyl methylcellulose and other cellulose derivatives, cyclopentane, hydroxypropyl methylcellulose and other cellulose derivatives). Dextrins (e.g., α-, β-, or γ-cyclodextrin and cyclodextrin derivatives); polyethylene glycol ethers or tetrahydrofurfuryl alcohol PEG ethers having an average molecular weight of about 200 to about 6000 (e.g., glycofurol, sold under the trade name Tetraglycol by BASF); surfactants (e.g., sodium lauryl sulfate, oleic acid, linoleic acid, glyceryl monooleate, lecithin, lysolecithin, deoxycholate, taurodeoxycholate, glycochenodeoxycholate, polyoxyethylene X-lauryl sulfate, glyceryl monooleate ... alkyl ethers (X is 9 to 20), sodium tauro-24,25-dihydrofusate, polyoxyethylene ether, polyoxyethylene sorbitan ester, 4-tert-octylphenoxypolyoxyethylene, N-lauryl-β-D-maltopyranoside, 1-dodecylazacycloheptan-2-one; amides (such as 2-pyrrolidone, 2-piperidone, caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide , polyvinyl pyrrolidone); esters (such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl octanoate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, caprolactone and its isomers, valerolactone and its isomers, β-butyrolactone and its isomers); and other solubilizers known in the art (such as dimethylacetamide, dimethyl isosorbide (Arlasolve DMI, ICI), N-methylpyrrolidone (Pharmasolve, ISP), monooctanoin, diethylene glycol monoethyl ether (Gattefosse commercially available under the trade name Transcutol)).

[0220] Preferred additional solubilizers include triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-1000, PEG 300, PEG 400, diethylene glycol monoethyl ether, dimethyl isosorbide, sorbitol, glycerol, triacetin, glycoforin, and propylene glycol. Typically, if the solubilizer is present, it is present in an amount of about 0.1% (w / v) to about 50% (w / v), about 1% (w / v) to about 40% (w / v), or about 2% (w / v) to about 25% (w / v). Additionally, the liquid pharmaceutical composition may comprise water, e.g., from about 2% (w / v) to about 99% (w / v), from about 10% (w / v) to about 95% (w / v), or from about 20% (w / v) to about 90% (w / v) of the liquid composition.

[0221] As previously mentioned, the composition may contain a preservative. Additionally or alternatively, the composition may be sterilized, which can be achieved by filtration sterilization, autoclaving, exposure to ionizing radiation (e.g., gamma rays, ultraviolet light), and chemical sterilization. In one embodiment, the sterility assurance level of the sterile composition is at least about 10 3 The resulting liquid composition is preferably stable at room temperature, for example, the therapeutic agent degrades by less than 5%, 4%, 3%, 2% or 1% by weight after storage at 20°C for 30 days or more preferably 6 months.

[0222] In addition, the formulation may also contain a sweetener or flavoring agent, exemplary sweeteners or flavoring agents include gum arabic syrup, acesulfame potassium, anethole, anise oil, aromatic elixir, aspartame, benzaldehyde, benzaldehyde elixir, cyclodextrin, caraway, caraway oil, cardamom oil, cardamom seeds, cardamom spirit, cardamom tincture, cherry juice, cherry syrup, cinnamon, cinnamon oil, cinnamon water, citric acid, citric acid syrup, clove oil, cocoa, cocoa syrup, coriander oil, glucose, eriodictyol, eriodictyol fluid extract, eriodictyol aromatic syrup, ethyl acetate, ethyl vanillin, anise oil, ginger, ginger fluid extract, ginger oleoresin, glucose, sucrose, maltodextrin, glycerin, licorice, licorice elixir, licorice extract, pure licorice extract, licorice fluid extract ointment, licorice syrup, honey, isoalcoholic elixir, lavender oil, lemon oil, lemon tincture, maltodextrin, maltose, mannitol, methyl salicylate, menthol, nutmeg oil, orange bitters elixir, orange bitters oil, orange blossom oil, orange blossom water, orange oil, bitter orange peel, sweet orange peel tincture, orange spirit, orange syrup, mint, mint oil, mint spirit, mint water, phenylethyl alcohol, raspberry juice, raspberry syrup, rosemary oil, rose oil, rose water, saccharin, saccharin calcium, saccharin sodium, sarsaparilla syrup, compound sarsaparilla, sorbitol solution, spearmint, spearmint oil, sucrose, sucralose, syrup, thyme oil, tolu balsam, tolu balsam syrup, wintergreen oil, vanilla, vanilla tincture, vanillin, wild cherry syrup, xylitol, or a combination thereof.

[0223] Additionally, the formulation may optionally contain a taste-masking agent, exemplary taste-masking agents include cyclodextrins, cyclodextrin emulsions, cyclodextrin particles, cyclodextrin complexes, or combinations thereof.

[0224] Dry powder composition

[0225] In certain embodiments, the NMDP compositions prepared according to the methods described herein can be designed for desired NMDP levels. For example, in one embodiment, when the microspheres are separated and deagglomerated by at least 77.8% of the deagglomerated particles (e.g., lactose monohydrate), the content of NMDP hydrochloride or its hydrate in the form of microspheres is about 22.2% w / w. In some embodiments, the final composition can then be introduced into a disposable dosage device (described below) to provide a dose of about 50 to about 2500 mg of NMDP hydrochloride in a single intranasal administration.

[0226] The pharmaceutical composition of the present disclosure can be contained in a disposable dosage unit for intranasal administration, providing a predetermined quantitative dose of NMDP or its salt. In one example of a disposable unit, a unit dose powder device (UDS) manufactured by Aptar Pharma. This device for powder spray can be used by non-medical professionals or patients or caregivers without medical training to systemically deliver small and accurately measured doses of NMDP preparations.

[0227] The present disclosure also relates to a dosage unit form (also referred to as a dosage unit device, dosage device or pharmaceutical device), in particular a disposable dosage unit form, for administering a single dose of the disclosed pharmaceutical composition to a subject intranasally, comprising an opioid receptor antagonist as an active ingredient, specifically NMDP or a pharmaceutically acceptable salt thereof (such as NMDP hydrochloride), wherein the dosage unit is loaded with a predetermined dose of the composition and provides a quantitative dose of the pharmaceutically active ingredient contained in the composition to the subject. In some embodiments, the dosage unit form is storage stable. In addition to the above-mentioned dosage unit device, the NMDP composition of the present disclosure can be administered using a syringe-driven device and a pump-driven spray nebulizer. The scope of the present disclosure also includes double-dose and multi-dose administration devices. In some embodiments, the NMDP contained in the dosage device of the present disclosure is retained in an amorphous form.

[0228] Compositions and dosage units thereof comprising NMDP or a pharmaceutically acceptable salt thereof as the active opioid receptor antagonist according to this aspect of the disclosure are also referred to herein as NMDP compositions and NMDP dosage units, respectively.

[0229] The NMDP compositions of the present disclosure are particularly useful for treating tinnitus or Meniere's disease. Treatment of tinnitus or Meniere's disease, as described herein, should be understood as alleviating or reversing the effects of tinnitus or Meniere's disease, and the symptoms associated therewith (e.g., hearing loss, ear pressure, etc.). The NMDP compositions of the present disclosure and their dosage units have been shown to have significant therapeutic effects.

[0230] In some embodiments, when the NMDP intranasal powder formulation according to the present disclosure is used for intranasal administration, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 50%, at least 60%, at least 70%, at least 80% or at least 86% of the NMDP particles reach the turbinate region. In some embodiments, when the NMDP intranasal powder formulation according to the present disclosure is used for intranasal administration, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 50%, at least 60%, at least 70%, at least 80% or at least 86% of the NMDP particles reach the ear region. In some embodiments, about 15%, about 20%, about 25%, about 30% or about 35% are located in the middle part of the olfactory region. In some embodiments, less than 10% of the NMDP particles are located in the nasal cavity, and less than 1% reach the lungs, thereby providing an effective amount of NMDP to the treated subject and improving the therapeutic effect.

[0231] Provided herein is a kit for treating / reversing tinnitus or Meniere's disease, comprising at least one dosage unit of the NMDP powder composition disclosed herein and instructions for use. The dosage unit may be a single dose, a double dose, or a multiple dose unit.

[0232] In one embodiment, a pharmaceutical composition in dry powder form for intranasal (nose-to-brain) N2B administration to a patient in need thereof comprises solid particles of NMDP and solid particles of a diluent, the pharmaceutical composition being substantially free of excipients other than the solid diluent, wherein at least 90% of the NMDP particles in the pharmaceutical composition have an average particle size of 10-30 microns, and less than 10% of the particles of the at least one active agent have an average particle size of about 5-50 microns, and the particles of the diluent have an average particle size of 50-200 microns.

[0233] In some embodiments, the composition may include a solid diluent such as lactose monohydrate or a functional analog of lactose. In another embodiment, a pharmaceutical composition in dry powder form for intranasal administration via the upper nasal mucosa (the turbinates and lymphoid tissue located at the back of the nasal cavity) for transmucosal systemic delivery comprises NMDP or a salt thereof having an average particle size in the range of 10-30 microns. In some embodiments, at least 90% of the particles thereof have an average particle size of not less than 5 microns and not more than 30 microns, and the average particle size of the diluent is in the range of 50-200 microns. As described above, the diluent is also used to prevent aggregation of dry powder particles containing NMDP or a salt thereof.

[0234] The compositions of the embodiments can be delivered by any nasal device known in the art, such as a pressurized device, a dry powder sprayer, or a bidirectional nasal device.Multi-dose devices and single-dose devices can be used.

[0235] The NMDP content of the compositions of the embodiments can be adjusted to provide the total drug dose required to achieve a therapeutic effect as a single dose in a single nostril. Repeated administration in a second nostril can double the amount of active substance. The stability of the compositions of the embodiments during storage can be determined under accelerated and ambient conditions.

[0236] In some embodiments, the compositions herein may comprise NMDP and a diluent (e.g., lactose or a functional analog of lactose) and be substantially free of other excipients (e.g., surfactants, lipid agents, solvents, or propellants). The solid diluent of the embodiments may be selected from lactose monohydrate or a functional analog of lactose monohydrate, such as lactose, cellulose and its derivatives, starch and its derivatives, glucose, sorbitol, mannitol, maltitol, xylitol, or a mixture thereof. The solid diluent may be lactose monohydrate.

[0237] Lactose may be present in the form of α-lactose monohydrate, anhydrous β-lactose, or amorphous lactose. The pharmaceutical composition of the embodiment may further comprise one or more pharmaceutically acceptable diluents, excipients, or both. The pharmaceutical composition of the embodiment may be prepared in the form of a powder, a simple powder mixture, powder microspheres, coated powder microspheres, liposomal dispersions, or a combination thereof.

[0238] In some embodiments of the present disclosure, the therapeutically effective amount of NMDP or a salt thereof is equivalent to about 10 mg, administered nasally in a single dose, to about 50 mg or 2500 mg of NMDP, optionally administered in several doses. In some embodiments, the therapeutically effective amount is equivalent to about 3, 4, 5, 6, 7 or 8 to about 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 80, 90, 100, 120, 150, 200 or 2500 mg of NMDP, respectively. In some embodiments, NMDP or a salt thereof is the only pharmaceutically active compound in the pharmaceutical composition. In some embodiments, the treatment methods according to the NMDP powder compositions and formulations of the present disclosure can result in a T of the plasma concentration-time curve of the NMDP in the patient. max Between about 0.13 and about 0.75 hours, for example, T max The treatment methods according to the NMDP powder compositions and formulations of the present disclosure can provide an average maximum plasma concentration of NMDP of about 20 pg / mL to about 25, 30, 40, 45, 50, 70, 90, 100, 120, 150, 180, 200, 250, 500, 1000 ng / mL or 20 ng / mL, 50, 100, 150 or 250 ng / mL within 15 minutes after administration. Treatment methods according to the NMDP powder compositions and formulations of the present disclosure can provide an average maximum concentration of NMDP in the otic structures of about 20 pg / mL to about 25, 30, 40, 45, 50, 70, 90, 100, 120, 150, 180, 200, 250, 500, 1000 ng / mL, or 20 ng / mL, 50, 100, 150, or 250 ng / mL within 15 minutes after administration.

[0239] Devices for intranasal delivery of the compositions of the embodiments can be designed to provide appropriate plume geometry and spray patterns for the initial and reservoir compositions. In some embodiments, these compositions can have a narrow particle size distribution with a median diameter between 5 and 50 microns.

[0240] Liposome composition

[0241] Provided herein is a method of administering NMDP or a salt thereof to a patient in need thereof, the method comprising intranasally administering a composition comprising a therapeutically effective amount of NMDP or a salt thereof, a phospholipid, one or more C2-C4 alcohols, and water, wherein the concentrations of the phospholipid and the one or more alcohols in the composition are 0.2 to 70% and 10 to 70% (by weight), respectively. In some embodiments, the water content of the composition is 10%, 20%, 30%, 40%, 50% or 60% (by weight), and the phospholipid forms vesicles in the composition. In some embodiments, the concentration of NMDP or a salt thereof is 0.5 to 25% (by weight).

[0242] Provided herein is an aqueous composition comprising NMDP or a salt thereof, comprising a phospholipid at a concentration of 0.2 to 50% (by weight) and in combination with one or more short-chain alcohols. In some embodiments, the weight concentration of water is at least 30% (by weight). In some embodiments, the weight concentration of the alcohol is in the range of 10 to 50% (by weight) and is suitable for use as an intranasal drug delivery vehicle. In some embodiments, the concentration of NMDP or a salt thereof in the composition is 0.5 to 25% (by weight).

[0243] Thus, in one aspect, described herein is a use of a vesicle composition comprising NMDP or a salt thereof, a phospholipid, one or more C2-C4 alcohols, and water for intranasal administration to treat one or more symptoms of tinnitus or Meniere's disease in a patient in need thereof. In some embodiments, the concentrations of the phospholipid and the one or more alcohols in the composition are 0.2 to 50% and 10 to 50% (by weight), respectively, and the water content of the composition is 10-50% (by weight). In some embodiments, the concentration of NMDP or a salt thereof is 0.5 to 25% (by weight).

[0244] Phospholipids suitable for preparing the compositions of the present disclosure include phosphatidylcholine (PC), hydrogenated phosphatidylcholine, phosphatidic acid (PA), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylglycerol (PPG) and phosphatidylinositol (PL). The chemical structures of phospholipids that can be used according to the present disclosure are described in U.S. Patent No. 4,614,730, which is incorporated herein by reference. Preferably, the concentration of phospholipids in the compositions of the present disclosure for intranasal delivery of NMDP to treat tinnitus or Meniere's disease is 0.5 to 15% (by weight).

[0245] The term "C2-C4 alcohol" as used herein refers to an alkanol containing two, three or four carbon atoms. The alcohols used in accordance with the present disclosure specifically include ethanol, 1-propanol, isopropanol and tert-butanol. In some embodiments, the concentration of ethanol in the composition is 1% to 20%, 30%, 40%, 50%, 60% or 70% (by weight). According to one embodiment of the present disclosure, the composition further comprises one or more water-miscible polyols, particularly glycols (1,2-glycols, such as ethylene glycol and propylene glycol), in a concentration of 1 to 30%, 40% or 50% (by weight).

[0246] The composition can be prepared by mixing the various components, namely water, phospholipids, one or more C2-C4 alcohols (possibly also one or more polyols), and NMDP or a salt thereof, under conditions that allow vesicle formation. In some embodiments, the composition can be conveniently prepared by dissolving the phospholipid in an alcohol (or alcohol / diol mixture), followed by adding NMDP or a salt thereof (in aqueous solution or solid form), and then adding water. Alternatively, a dispersion of the phospholipid and NMDP or a salt thereof in water is prepared, to which the alcohol is added, optionally together with a polyol (e.g., a mixture of ethanol and propylene glycol), under stirring (possibly under heating).

[0247] Alternatively, freeze-dried lipid vesicles encapsulating the active ingredient may be prepared first and then dispersed in a mixture of water, a C2-C4 alcohol and optionally a polyol.

[0248] In some embodiments, the size of the vesicles can range from 50 nm to several microns, more specifically up to 5 μm. In some embodiments, the compositions according to the present disclosure may include additional excipients known in the art, such as surfactants, preservatives, thickeners, solubilizers, binders, antioxidants, buffers, viscosity and absorption enhancers, and agents capable of adjusting the pH and osmotic pressure of the formulation. Additional excipients that can be used with the compositions and methods herein are described elsewhere in this specification.

[0249] In another aspect, the present disclosure provides a method of administering an active pharmaceutical ingredient to a patient in need thereof, comprising intranasally administering a vesicle composition comprising a therapeutically effective amount of the ingredient, a phospholipid, one or more C2-C4 alcohols, and water, wherein the concentrations of the phospholipid and the one or more alcohols in the composition are 0.2 to 10% and 12 to 30% (by weight), respectively, and the water content of the composition is not less than 20%, preferably not less than 30% (by weight).

[0250] Sustained-release formulations

[0251] In certain aspects, the present disclosure describes a method and dosage form for sustained delivery of NMDP for treating tinnitus or symptoms associated with Meniere's disease in a mammalian subject. In some embodiments, it is contemplated that a mucosal delivery formulation of NMDP comprises one or more mucosal delivery enhancers, wherein the release of the NMDP dose is substantially normalized and / or sustained for an effective delivery time range of about 0.1 to about 2.0 hours; about 0.4 to about 1.5 hours; about 0.7 to about 1.5 hours; or about 0.8 to about 1.0 hours after mucosal administration. Repeated administration of exogenous NMDP using the methods and compositions of the present disclosure can facilitate sustained release of the NMDP.

[0252] In some embodiments, the mucosal delivery enhancers disclosed herein can effectively improve delivery, for example, increase the maximum plasma concentration (Cmax ) or C in ear structures max , to enhance the therapeutic activity of NMDP administered transmucosally. Another factor affecting the therapeutic activity of NMDP in treating tinnitus or Meniere's disease in ear structures is residence time (RT). In some embodiments, a sustained release enhancer is combined with an intranasal delivery enhancer to increase C max and increasing the residence time (RT) of NMDP in the otic structures. It is contemplated herein to increase the residence time at the mucosal delivery site (e.g., nasal mucosa), at the treatment site (e.g., otic structures), and / or in the systemic circulation. In some embodiments, for example, the polymeric delivery vehicles and other agents and methods of producing sustained-release enhanced formulations disclosed herein may include polyethylene glycol (PEG).

[0253] Semisolid and viscous gel preparations

[0254] In some methods contemplated herein, the intranasal NMDP nasal pharmaceutical composition is applied to the lateral wall of the nasal cavity of each nostril (opposite to the nasal septum), preferably the cartilage of the lateral wall of each nasal cavity is locally located in the middle of the lateral wall (opposite to the nasal septum) to the portion immediately below the top. After the nasal pharmaceutical composition is deposited in each nostril of the nasal cavity, the external nose is gently and carefully squeezed and / or the subject is rubbed so that the deposited nasal pharmaceutical composition is ready to release NMDP continuously by maintaining contact with the nasal mucosa. The typical NMDP nasal pharmaceutical composition dose deposited during nasal administration is about 50 to about 150 microliters per nasal cavity, preferably about 100 microliters per nasal cavity.

[0255] In practicing the methods of the present disclosure, about 50 microliters to about 150 microliters of the pharmaceutical composition of the present disclosure can be administered to each nostril of the subject daily, for example, for 1, 2, 3, 4 weeks, three, four, five, six, seven, eight months or longer, or intermittently (e.g., for more than two, three, four, five or six months in a row), or once, two or three times a week, or as needed for the hearing impairment.

[0256] The methods disclosed herein include dosage concentrations of intranasal NMDP compositions, number of daily administrations, durations of treatment, non-oral methods, and in some embodiments, pre-filled multi-dose applicator systems that deliver effective dosage levels of NMDP or mixtures thereof in intranasal compositions that can be administered any number of times daily, weekly, monthly, or yearly and deliver effective amounts of NMDP or mixtures thereof.

[0257] In some embodiments, the viscosity of the nasal pharmaceutical composition of the present disclosure is at least about 500 cps (e.g., cP), and the thixotropy associated with some of the novel nasal pharmaceutical compositions prior to administration is within a range of about 500 cps to about 100,000 cps. In some embodiments, the viscosity is from about 1000 cps to about 75,000 cps, from about 2500 cps to about 50,000 cps, and the thixotropy associated with some of the novel nasal pharmaceutical compositions prior to administration or pump operation may be within a range of about 5,000 cps to about 25,000 cps. In some embodiments, the viscosity of the composition is from about 10 to about 100 cps, from about 10 to about 90 cps, from about 10 to about 80 cps, from about 10 to about 70 cps, from about 10 to about 60 cps, or from about 10 to about 50 cps.

[0258] In some embodiments, the applicator system of the present disclosure is, for example, an airless fluid, a dip tube fluid dispensing system or pump, or any other system suitable for implementing the methods of the present disclosure. The applicator system or pump includes a chamber pre-filled with multiple doses of the intranasal NMDP gel of the present disclosure, which is closed by, for example, a drive nozzle. The drive nozzle may include an outlet channel and a tip, wherein the drive nozzle is designed to: (a) continuously deliver a uniform volume of the intranasal NMDP gel of the present disclosure when applied around the patient's nasal cavity; (b) be constructed to conform to the inner surface of the user's nostrils to deposit at each designated location in the patient's nostrils as contemplated by the new methods and teachings of the present disclosure. Preferably, when inserted into the nasal cavity, the design of the pump is constructed to ensure that the nasal tip is correctly positioned in the nasal cavity, thereby releasing the gel to the appropriate location in the nasal cavity when it is dispensed.

[0259] Oily gels, emulsions, or creams can be applied using a properly designed and secure dispenser tip about 1 inch inside the nostril opening (non-air area) to reach the nasal cavity and adhere to the container. The tip is preferably rounded to prevent damage. The nose is then massaged to spread the composition into a thin film within the nasal cavity, which will help the active ingredient be absorbed by the mucosal tissue.

[0260] Examples of pre-filled multi-dose applicator systems include: (a) the COMOD system from Ursatec, Verpackung-GmbH, Schillerstr. Wendel, Germany; (b) the digital airless applicator system from Airlessystems, RD14927380 Charleval, France or 303 North, Congers, NY 10950, USA; (c) the nasal applicator from Neopac, The Tube, Hoffmann Neopac AG, Burgdorfstrasse 22, Postfach, 3672 Oberdiessbach, Switzerland; or (d) a cannabinoid pharmaceutical composition.

[0261] Preferably, the intranasal NMDP pharmaceutical composition is filled into a preservative-free airless multi-dose device that is capable of accurately delivering a higher viscosity NMDP pharmaceutical composition.

[0262] According to certain embodiments, the composition comprises: (1) an NMDP therapeutically active ingredient; (2) an oily vehicle; and (3) a wetting agent, or a mixture of a wetting agent and / or a pharmaceutically acceptable surfactant, or a mixture of surfactants.

[0263] According to certain embodiments, the oily carrier is one or more lipids that are generally recognized as pharmaceutically acceptable and safe.

[0264] According to certain embodiments, the oily carrier is selected from the group consisting of pharmaceutically acceptable vegetable oils, monoglycerides, diglycerides, sucrose acetate isobutyrate (SAIB), synthetic triglycerides, and combinations thereof. According to a specific embodiment, the pharmaceutically acceptable vegetable oil is selected from the group consisting of sweet almond oil (Prunus dulcis), virgin almond oil (Prunus amid glarus), aloe vera oil (Aroebabadensis), almond oil (fruit pandanus Armenia car), argan oil (are theCalifornia RY labor), avocado oil (PERE Seah Americana), apricot oil (fruity Taunus Armeniacar), mango oil (amra oil) (preamble Rica operational during the day lease), borage oil (swabbing Opie during the day lease), black seed oil (age Gela sativa), carrot oil (DowSyracuse Caro L), coconut oil (nose Syracuse nusi Pera), corn oil, cucumber oil (Cucumis tiva), tea seed oil (tea ulmu gras oil) (hydroxy-no kapuseu Wiig Tia Taunus), emu oil (draw Myfunny nobae-Hall Grandi kids), evening primrose oil (moon yikkot oil) (Oe BrunoTerra Bien Nice), linseed oil (rineom Wuxi City Tatiana stopped), grape seed oil (VitusBeanie Blow), hazelnut oil (ahbekeu Kana), camellia oil (Yo bar purified oil) (deep diamondcyano difference norbornene-cis), Moringa oil (Moringa five days Blow to raise) (Moringa), eucalyptus oil (e Lula oil) ('s Klee Rocca Ria non LEA), wheat germ oil, treetikum No Les, macadamia nut oil (Macadamia Terni polyamic), cantaloupe oil (Kubu Miss melon), Maersk oil (Abel Moss cheoseu Moss primary Tooth),Mustard oil (Azadi lakhother indica), olive oil (oleic Ah Europa), peach kernel oil (fruity Taunus Pere Chicago), peanut oil (arachis hypo geah), pomegranate oil, Fu Nika Gras natum, program soral Leah oil (program soral Leah Cora Come polyamic), rose essential oil (Oe Bruno Terra BienNeedle), papaya seed oil (K Rica papaya), rosehip oil (Rosa Ruby-basedlabor), safflower oil, refined sesame oil (Cesar stopped Indy Com), the living tree oil (Hippo wave Lam Noideath), soybean oil (Both oils) (Soya Heath the blood), sunflower oil (Heli no tooth should Taunus), sweet almond oil (fruit pandanus Ami month Russ Barr moon Syracuse), sweet cherry kernel oil (fruity Taunus Oh Away), walnut oil (jugeul Lance Reg A), Watermelon seed oil (citrus vulgaris).

[0265] According to certain embodiments, the oily vehicle comprises castor oil and / or sesame oil and / or SAIB (sucrose acetate isobutyrate).

[0266] According to a particular embodiment, the wetting agent or a mixture of wetting agents and / or pharmaceutically acceptable surfactants or a mixture of surfactants is selected from the group consisting of: polysorbates, polyoxyethylene hydrogenated vegetable oils, polyoxyethylene vegetable oils; polyoxyethylene sorbitan fatty acid esters; polyoxyethylene-polyoxypropylene block copolymers; polyglycerol fatty acid esters; polyoxyethylene glycerides; polyoxyethylene sterols or derivatives or analogs thereof; reaction mixtures of polyols with at least one of fatty acids, glycerides, vegetable oils, hydrogenated vegetable oils, fractionated oils and sterols; tocopheryl polyethylene glycol succinate; sugar esters; sugar ethers; sucrose glycerides; alkyl glucosides; alkyl Maltoside; alkyl thioglycoside; lauryl macrogol glyceride; polyoxyethylene alkyl ether; polyoxyethylene alkylphenol; polyethylene glycol fatty acid ester; polyethylene glycol glycerol fatty acid ester; polyoxyethylene sorbitan fatty acid ester; polyoxyethylene-polyoxypropylene block copolymer (such as poloxamer-108, 188, 217, 238, 288, 338, 407, 124, 182, 183, 212, 331 or 335, or a combination thereof); ionic hydrophilic surfactant (such as sodium lauryl sulfate or docusate sodium); bile acid; cholic acid; deoxycholic acid; chenodeoxycholic acid; salts thereof, salts thereof and mixtures thereof.

[0267] According to certain embodiments, the composition comprises a rheology modifier such as colloidal silica, silicates, alumina, high molecular weight polymers or solid / waxy materials, beeswax, alumina, silica, silicates and high melting point waxes and / or cetearyl alcohol.

[0268] According to certain embodiments, the composition further comprises a mineral, an osmotic pressure supplement, a thickener and / or a hydrophilic polymer. According to certain embodiments, the hydrophilic polymer is selected from the group consisting of hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sodium carboxymethyl cellulose (sodium CMC), sodium carboxymethyl cellulose (sodium CMC) and microcrystalline cellulose (MCC), xanthan gum, guar gum, gum arabic, natural gums (such as tragacanth gum), corn starch, potato starch and starches. According to a particular embodiment, the surfactant is selected from the group consisting of ethylene glycol distearate, sorbitan trioleate, propylene glycol isostearate, ethylene glycol stearate, sorbitan sesquioleate, lecithin, sorbitan oleate, sorbitan monostearate (NF), sorbitan stearate, sorbitan isostearate, stearate-2, oleate-2, glyceryl laurate, cetyl alcohol-2, PEG-30 dipolyhydroxystearate, glyceryl stearate SE, sorbitan PEG-8 sorbitan laurate, sorbitan laurate, sorbitan monolaurate, macrogol glyceryl oleate, laureth-4, PEG-7 glyceryl cocoate, PEG-20 almond glycerides, polyoxyethylene glyceryl stearate, P EG-25 Hydrogenated Castor Oil, Stearamide MEA, Glyceryl Stearate (and) PEG-100 Stearate, Polysorbate 85, PEG-7 Olivate, Cetearyl Glucoside, Stearamide MEA, PEG-10, Oleth-10 / Polyoxyethylene 10 Oleyl Ether (NF), Ceteth-10, PEG-8 Laurate, Cocamide MEA, Polysorbate 60 (NF), Polysorbate 60, Polysorbate 80, Isostearate 20, PEG-60 Almond Glycerides, PEG-20 Methyl Glucose Sesquistearate, Ceteareth-20, Oleate-20, Steareth-20, Steareth-21, Ceteth-20, and Steareth-100.

[0269] Excipients and other ingredients in the formulation

[0270] In various aspects of the present disclosure, there are provided improved nasal mucosal delivery formulations and methods that allow NMDP and / or other therapeutic agents to be delivered between the administration site and one or more selected target sites through a mucosal barrier (e.g., mucosal surface). Certain formulations may be specifically adapted to selected target cells, tissues, or organs, or even specific disease states. In other aspects of the present disclosure, improved intranasal delivery formulations and methods can achieve efficient, selective endocytosis or transcytosis of NMDP along defined intracellular or intercellular pathways. As understood herein, NMDP can be efficiently loaded in a carrier or other delivery vehicle at an effective concentration, and then maintained in a stable form during administration (e.g., when applied to the nasal mucosa) and / or in the process of reaching the drug action target site (e.g., bloodstream or specific tissues, organs, or extracellular compartments) through one or more intracellular compartments and / or membranes. NMDP can be provided by a delivery vehicle or otherwise modified (e.g., in the form of a prodrug), wherein the release or activation of NMDP is triggered by physiological stimuli (e.g., pH changes, lysosomal enzymes, etc.). In certain aspects, NMDP has no pharmacological activity before reaching its active target site. NMDP and other formulation components are non-toxic (or their toxicity is reduced to an acceptable level) and non-immunogenic. In this context, carriers and other formulation components are typically selected based on their ability to rapidly degrade and / or be excreted under physiological conditions. At the same time, the formulation has chemical and physical stability in dosage form to achieve effective storage.

[0271] In the mucosal delivery formulations and methods of the present disclosure, NMDP is typically combined or co-administered with a carrier or vehicle suitable for mucosal delivery. As used herein, the term "carrier" includes pharmaceutically acceptable solid or liquid fillers, diluents, or encapsulating materials. In this context, the carrier can be a mucosal delivery enhancer.

[0272] The aqueous liquid carrier may contain pharmaceutically acceptable additives such as acidifiers, alkalizers, antimicrobial preservatives, antioxidants, buffers, chelating agents, complexing agents, solubilizers, humectants, solvents, suspending agents and / or viscosity-increasing agents (such as thickeners), tonicity agents, wetting agents or other biocompatible materials. As disclosed herein, humectants include, but are not limited to, propylene glycol, glycerol, triacetin, polyols, polymeric polyols, lactic acid and urea. In the present disclosure, the pharmaceutical formulation may contain one humectant or any combination or mixture of more than one humectant.

[0273] The solubilizers included in the composition herein may include cyclodextrin, hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin and methyl-β-cyclodextrin. Such solubilizers can be used alone in the pharmaceutical formulation, or used in any mixture or combination with more than one solubilizer. Examples of materials that can be used as pharmaceutically acceptable carriers include sugars (such as lactose, glucose and sucrose); starches (such as corn starch and potato starch); cellulose and its derivatives (such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate); tragacanth powder; malt; gelatin; talc; excipients (such as cocoa butter and suppository wax); oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil); glycols (such as propylene glycol); polyols (such as glycerol, sorbitol, mannitol and polyethylene glycol); esters (such as ethyl oleate and ethyl laurate); agar; buffers (such as magnesium hydroxide and aluminum hydroxide); alginic acid; pyrogen-free water; isotonic saline, acetate, glycine, histidine, arginine, glutamic acid, lysine, methionine, lactate, formate and glycolate; Ringer's solution, ethanol and phosphate buffer solutions, and other nontoxic compatible substances used in pharmaceutical formulations.

[0274] In some embodiments, the pharmaceutical formulations described herein for transmucosal (e.g., intranasal) delivery of NMDP or its salts may include any one buffer or any combination or mixture of more than one buffer. The pKa range of the buffer may be from about 5 to about 9, or from about 6 to about 8. Depending on the needs of the formulation personnel, wetting agents, emulsifiers, and lubricants (e.g., sodium lauryl sulfate and magnesium stearate), as well as colorants, release agents, coating agents, sweeteners, flavorings and fragrances, preservatives, and antioxidants may also be present in the composition. Examples of pharmaceutically acceptable antioxidants include water-soluble antioxidants (such as ascorbic acid, cysteine hydrochloride, sodium bisulfite, sodium metabisulfite, sodium sulfite, etc.); oil-soluble antioxidants (such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.); and metal chelators (such as citric acid, ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), sorbitol, tartaric acid, phosphoric acid, etc.). According to the present disclosure, the pharmaceutical formulation may contain any one chelating agent or any mixture or combination of chelating agents. The amount of active ingredient combined with the carrier material to prepare a single-dose dosage form will vary depending on the specific mode of administration.

[0275] In the mucosal delivery compositions and methods of the present disclosure, various mucosal delivery enhancers are used, which can enhance the delivery of NMDP into or across mucosal surfaces. In this regard, the delivery of NMDP across mucosal epithelium can occur "transcellularly" or "paracellularly". The contribution of these pathways to the total flux and bioavailability of NMDP depends on the mucosal environment, the physicochemical properties of the active ingredient, and the characteristics of the mucosal epithelium. In some embodiments, the methods and compositions of the present disclosure can significantly enhance the entry and transmucosal transport of NMDP or its salts through the paracellular pathway. In some embodiments, the methods and compositions of the present disclosure can significantly enhance the entry and transmucosal transport of NMDP or its salts through the transcellular pathway. In some embodiments, the methods and compositions of the present disclosure can significantly enhance the entry and transmucosal transport of NMDP or its salts through the paracellular pathway, the transcellular pathway, or alternately through the above-mentioned pathways in a single method or composition, significantly enhancing the entry and transmucosal transport of NMDP or its salts.

[0276] As used herein, mucosal delivery enhancers include agents that enhance or otherwise modulate the release or solubility (e.g., from a formulation delivery vehicle), diffusion rate, permeability and timing, uptake, residence time, stability, effective half-life, peak or sustained concentration levels, clearance, and other desired mucosal delivery characteristics of an NMDP or other biologically active compound (e.g., as measured at the site of delivery or a selected active target site (e.g., bloodstream or central nervous system). Thus, enhancement of mucosal delivery can be achieved by any one or more of a variety of mechanisms, such as by increasing the diffusion, transport, persistence, or stability of the NMDP, increasing membrane fluidity, modulating the availability or action of calcium and other ions that regulate intracellular or paracellular permeation, solubilizing mucosal membrane components (e.g., lipids), altering non-protein and protein sulfhydryl levels in mucosal tissue, increasing water flux across mucosal surfaces, modulating epithelial junction physiology, reducing the viscosity of the mucus covering the mucosal epithelium, reducing the rate of mucociliary clearance, and other mechanisms.

[0277] As used herein, "mucosal effective amount of NMDP" refers to the effective mucosal delivery of NMDP to the target site (i.e., ear structure) to produce drug activity in a subject in need, which may involve a variety of delivery or transfer pathways. For example, NMDP or a salt thereof can pass through the gaps (such as spaces) between mucosal cells and reach the adjacent blood vessel walls in the Eustachian tube, while through another approach, the agent can be passively or actively taken up (i.e., internalized) into mucosal cells to play a role within the cell, or be expelled (such as released) or transported out of the cell to reach a secondary target site (such as the tympanic membrane). The methods and compositions of the present disclosure can promote the transport of NMDP or its salt along one or more such alternative (transcellular or paracellular) pathways, or can act directly on mucosal tissue or proximal vascular tissue to promote the absorption or penetration of NMDP or its salt. In this context, the promotion of absorption or penetration is not limited to these mechanisms.

[0278] Suitable surfactants that can be used according to the present disclosure include ionic, nonionic or amphoteric surfactants. More specifically, hydrophilic surfactants (such as Tweens, Tween 80, Myrij, Brij, Labrasol, etc.) or lipophilic surfactants (such as Span 20, Span 60, Myrij, Arlacel83, etc.) can be suitably used, preferably in a concentration range of 0-25% (w / w).

[0279] Suitable preservatives that can be used with the formulations of the present invention include, for example, benzyl alcohol, parabens, chlorobutanol, benzalkonium chloride, and combinations thereof. Some examples of antioxidants include tocopherol, butylated hydroxytoluene, sodium metabisulfite, potassium metabisulfite, ascorbyl palmitate, and the like. The concentration of these preservatives and antioxidants in the formulation can be from about 0.001% to about 5% (w / w).

[0280] With respect to buffers, the nasal delivery system may include a buffer for maintaining the formulation at approximately pH 7.0. Of course, the specific buffer may vary depending on the specific nasal delivery system used and the specific active molecule selected. Suitable buffers for use in the present disclosure include, for example, acetate, citrate, glutamine, carbonate, and phosphate buffers, and combinations thereof. The pharmaceutical formulations of the present disclosure may include a pH adjuster.

[0281] About thickening agent, the viscosity of the disclosed preparation can be maintained at the desired level using pharmaceutically acceptable thickening agent. The thickening agent that can be added into the disclosed composition includes for example methylcellulose, xanthan gum, tragacanth gum, adhesive, guar gum, carboxymethyl cellulose, hydroxypropyl cellulose, carbomer, polyvinyl alcohol, alginate, gum arabic, chitosan, mucoadhesive polymer system (such as polyacrylate), cellulose derivative, hyaluronic acid, hyaluronic acid derivative, chitin, collagen, pectin, starch, polyethylene glycol, sulfated polysaccharide, carrageenan, sodium alginate, gelatin, pectin and combination thereof. The required concentration of thickening agent will depend on selected reagent and required viscosity.

[0282] The composition may also include gel-forming or bioadhesive compounds such as carbomers, alginates, scleroglucans, cellulose derivatives, starches, albumin, pluronic gel, diethylaminoethyl (DEAE)-dextran, polycarbophil, hyaluronic acid, hyaluronates, starches, gelatin, collagen, and the like. The composition may also be incorporated into water-in-oil creams, oil-in-water creams, hydrophilic or lipophilic ointments, gels, and other semisolid matrices. The composition may be delivered to the nasal cavity in the form of drops, mists, aerosols, or instillations using a dropper, a dedicated device, a vaporizer, a vaporizer, or the like.

[0283] The formulations of the present disclosure may also include agents such as tolerance enhancers to reduce or prevent drying of mucosal membranes and to prevent irritation thereof.

[0284] In some embodiments, NMDP or its salt can be made into a viscous matrix by adding commonly used ingredients (such as natural gums, cellulose and its derivatives, acrylic polymers (such as carbomer) and vinyl polymers (polyvinyl pyrrolidone), scleroglucan, xylan, alginate, calcium alginate, hyaluronate, collagen salt, starch gel, gelatin system, chitosan carrier) to the above-mentioned delivery system.

[0285] In some embodiments, the compositions disclosed herein may further comprise benzalkonium chloride (BKC) and / or ethylenediaminetetraacetic acid (EDTA). In some embodiments, the compositions may comprise, for example, 0.01, 0.05, 0.1, 0.2, 0.3, or 0.5% (w / v) BKC. In some embodiments, the compositions may comprise 0.01, 0.05, 0.1, 0.2, 0.3, or 0.5% (w / v) EDTA.

[0286] Although the absorption promoting mechanisms of different mucosal delivery enhancers in the present disclosure may be different, useful agents in this context will not have a substantial adverse effect on mucosal tissue and will be selected based on the physicochemical properties of the specific NMDP or other active or delivery enhancer. In this context, delivery enhancers that increase mucosal tissue permeability or permeability will generally result in certain changes in the mucosal protective permeability barrier. In order for such delivery enhancers to be valuable in the present disclosure, it is generally expected that any significant changes in mucosal permeability can be reversible within a timeframe suitable for the desired drug delivery duration. In addition, during long-term use, the mucosal barrier properties should not produce substantial cumulative toxicity or any permanent harmful changes.

[0287] In some aspects of the present disclosure, the absorption enhancer used in conjunction with NMDP or formulated in combination is selected from hydrophilic small molecules, including but not limited to dimethyl sulfoxide (DMSO), dimethylformamide, ethanol, propylene glycol and 2-pyrrolidone. Alternatively, long-chain amphiphilic molecules (such as deacyl methyl sulfoxide, azone, sodium lauryl sulfate, oleic acid and bile salts) can be used to enhance the mucosal permeability of NMDP. In other aspects, surfactants (such as polysorbate) are used as auxiliary compounds, processing agents or formulation additives to enhance the intranasal delivery of NMDP. Reagents such as DMSO, polyethylene glycol and ethanol, if present in a sufficiently high concentration in a delivery environment (such as by pre-administration or incorporation into a therapeutic preparation), can enter the aqueous phase of the mucosa and change its solubilizing properties, thereby enhancing the distribution of NMDP from a vehicle (such as treatment or pharmaceutical preparation) to the mucosa.

[0288] Other mucosal delivery enhancers useful in the coordinated administration and processing methods and combined formulations include, but are not limited to, mixed micelles; enamines; nitric oxide donors (such as S-nitroso-N-acetyl-DL-penicillamine, NOR1, NOR4 - which are preferably co-administered with NO scavengers such as carboxy-PITO or diclofenac sodium); sodium salicylate; acetoacetin (such as glycerol-1,3-diacetoacetate or 1,2-isopropylidene glycerol-3-acetoacetate); and other release-diffusion or intraepithelial / transepithelial permeation enhancers that are physiologically compatible with mucosal delivery.

[0289] Other absorption enhancers can be selected from a variety of carriers, matrices and excipients that enhance NMDP mucosal delivery, stability, activity or transepithelial permeability. These include cyclodextrins (such as β-cyclodextrin) and β-cyclodextrin derivatives (such as hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, methyl-β-cyclodextrin and seven (2,6-di-O-methyl-β-cyclodextrin)). These compounds are optionally conjugated to one or more active ingredients and further optionally formulated in an oily matrix to enhance the bioavailability of the glucose-regulating peptide contained in the mucosal preparation of the present invention. Other absorption enhancers suitable for mucosal delivery include medium-chain fatty acids, including monoglycerides and diglycerides (such as sodium caprate-coconut oil extract, Capmul) and triglycerides (such as starch dextrin, Estaram 299, Miglyol 810).

[0290] The mucosal treatment and prevention compositions of the present disclosure can be supplemented with any suitable penetration enhancer to promote the absorption, diffusion or penetration of NMDP across the mucosal barrier. The penetration enhancer can be any pharmaceutically acceptable such agent. Therefore, in a more detailed aspect of the present disclosure, there is provided a composition incorporating one or more penetration enhancers selected from the following: sodium salicylate and salicylic acid derivatives (acetylsalicylate, choline salicylate, salicylamide, etc.); amino acids and their salts (such as monoaminocarboxylic acids, such as glycine, alanine, phenylalanine, proline, hydroxyproline, etc.; hydroxyamino acids, such as serine; acidic amino acids, such as aspartic acid, glutamic acid, etc.; and basic amino acids, such as lysine, etc. - including alkali metal or alkaline earth metal salts thereof); and N-acetylamino acids (N-acetylalanine, N-acetylphenylalanine, N-acetylserine, N-acetylglycine, N-acetylysine, N-acetylglutamic acid, N-acetylproline, N-acetylhydroxyproline, etc.) and their salts (alkali metal salts and alkaline earth metal salts). The methods and compositions of the present disclosure also provide substances commonly used as emulsifiers (such as sodium lauryl phosphate, sodium lauryl sulfate, sodium myristyl sulfate, polyoxyethylene alkyl ethers, polyoxyethylene alkyl esters, etc.), caproic acid, lactic acid, malic acid and citric acid and their alkali metal salts, pyrrolidone carboxylic acid, alkylpyrrolidone carboxylic acid esters, N-alkylpyrrolidone, proline acyl esters, etc. as penetration enhancers.

[0291] It is expected that a range of components and additives can be used in the methods and formulations of the present disclosure. Examples of such solubilizing agents are cyclodextrins (CDs) and their derivatives. These CDs have been found to bind to hydrophobic regions of proteins in a manner that significantly inhibits aggregation. This inhibition is selective for both the CDs and the proteins involved. This selective inhibition of protein aggregation provides additional advantages in the intranasal delivery methods and compositions of the present disclosure. Other agents used in this context include CD dimers, trimers and tetramers with different geometric structures controlled by linkers, peptides, peptide derivatives, analogs and peptide mimetics to selectively block protein-protein interactions. In one aspect, the specific binding of reported CD polymers to hydrophobic side chains is extended to proteins by using peptides and peptide mimetics that similarly block protein aggregation. A wide range of suitable methods and anti-aggregating agents can be incorporated into the compositions and procedures contemplated herein.

[0292] Charge modification and pH control agents and methods

[0293] In order to improve the transport properties of bioactive agents (including NMDP) or other active macromolecules and small molecule drugs to enhance their delivery across hydrophobic mucosal barriers, the compositions herein may also use reagents for "charge modification" of the selected bioactive agents or delivery enhancers described herein. In this regard, the relative permeability of macromolecules is generally related to their partition coefficient. The degree of ionization of the molecule (depending on the pKa of the molecule and the pH of the mucosal membrane surface) also affects its permeability. As described herein, when bioactive agents including NMDP and its analogs are used for mucosal delivery, their penetration and distribution can be promoted by charge changes or charge distribution of the active agent or permeabilizing agent, which can be achieved by, for example, changing the charged functional group, adjusting the pH of the solution of the delivery vehicle or the delivery active agent (or its precursor), or by co-administering charge or pH change reagents with the active agent (or precursor).

[0294] Based on these general teachings, mucosal delivery of charged macromolecular species, including NMDPs and other bioactive peptides and proteins, in the methods and compositions of the present disclosure can be significantly improved when the active agent is delivered to the mucosal surface in a substantially non-ionized or neutrally charged state.

[0295] Certain compositions comprising NMDP salts (NMDP(s)) and one or more other bioactive peptide and protein components of mucosal formulations disclosed herein may be charge modified to increase the positive charge density of the peptide or protein. These modifications also extend to the cationization of the peptide and protein conjugates, carriers, and other delivery formats disclosed herein. Cationization provides a convenient means for altering the biodistribution and transport properties of proteins and macromolecules disclosed herein. Cationization should be performed in a manner that substantially retains the biological activity of the active agent and limits potential side effects (including tissue damage and toxicity).

[0296] In some embodiments, the composition may include a buffer solution. A "buffer" is generally used to maintain the pH of a solution at a near-constant value. Examples of commonly used buffer salts include the following: glutamate, acetate, citrate, glycine, histidine, arginine, lysine, methionine, lactate, formate, glycolate, tartrate, phosphate, and mixtures thereof.

[0297] Mucolytic and mucus-clearing agents and methods

[0298] In some embodiments, the compositions and methods for mucosal delivery (such as intranasal delivery of NMDP or its salt) herein may optionally incorporate an effective mucolytic or mucus-clearing agent, which is used to degrade, dilute or clear mucus on the surface of the intranasal mucosa to promote the absorption of the intranasal administration composition. In some embodiments, mucolytic or mucus-clearing agents are co-administered as auxiliary compounds to enhance the intranasal delivery of biologically active agents. Alternatively, an effective amount of a mucolytic or mucus-clearing agent is incorporated into the multi-step processing method of the present disclosure as a processing agent, or is incorporated into the combined formulation of the present disclosure as an additive to provide an improved formulation that enhances the intranasal delivery of biological therapeutic compounds by reducing the barrier effect of intranasal mucus.

[0299] A variety of mucolytic or mucus-clearing agents can be incorporated into the methods and compositions of the present disclosure. Based on their mechanism of action, mucolytic and mucus-clearing agents can generally be divided into the following categories: proteases that cleave the mucin glycoprotein protein core (e.g., pronase, papain); sulfhydryl compounds that split mucin disulfide bonds; and detergents that disrupt non-covalent bonds in mucus (e.g., Triton X-100, Tween 20). Other compounds in this context include, but are not limited to, bile salts and surfactants, such as sodium deoxycholate, sodium taurodeoxycholate, sodium glycocholate, and lysophosphatidylcholine.

[0300] The order of effectiveness of bile salts in causing mucus structural disruption is deoxycholate > taurocholate > glycocholate. According to the methods of the present disclosure, other effective agents for reducing mucus viscosity or adhesion to enhance intranasal delivery include, for example, short-chain fatty acids, and mucolytic agents that function by chelation, such as N-acyl collagen peptides, bile acids, and saponins (the latter in part by chelating Ca, which plays an important role in maintaining the structure of the mucus layer). 2+ and / or Mg 2+ function).

[0301] Other mucolytic agents used in the methods and compositions of the present disclosure include N-acetyl-L-cysteine (ACS), a potent mucolytic agent that reduces the viscosity and adhesiveness of bronchopulmonary mucus and has been reported to modestly increase the nasal bioavailability of human growth hormone in anesthetized rats (from 7.5% to 12.2%). These and other mucolytic or mucus-clearing agents are placed in contact with the nasal mucosa, typically at concentrations ranging from about 0.2 to about 20 mM, and act synergistically with the administration of the bioactive agent to reduce the polar viscosity and / or elasticity of intranasal mucus.

[0302] Other mucolytic or mucus-clearing agents can be selected from a range of glycosidases, which cleave glycosidic bonds within mucus glycoproteins. α-Amylase and β-amylase are representatives of this class of enzymes, although their mucolytic activity may be limited. In contrast, bacterial glycosidases allow these microorganisms to penetrate the host's mucus layer.

[0303] Non-ionic detergents can also generally act as mucolytic or mucus-clearing agents when used in combination with the bioactive agents described in the present disclosure.

[0304] Cilia inhibition reagents and methods

[0305] Because the self-cleaning ability of certain mucosal tissues, such as the nasal mucosa, through mucociliary clearance is an essential protective function (e.g., removal of dust, allergens, and bacteria), it is important to understand that mucosal-administered drugs should not substantially impair this function. Mucociliary transport in the respiratory tract is a particularly important defense mechanism against infection. To achieve this function, the beating of cilia in the nasal cavity and airways pushes the mucus layer along the mucosa to remove inhaled particles and microorganisms.

[0306] In some embodiments, the methods and compositions of the present disclosure may be incorporated with a cilium inhibitory agent to increase the residence time of a mucosal (e.g., intranasal) administration formulation comprising NMDP and other bioactive agents disclosed herein. In particular, in certain aspects, by co-administering or combining one or more cilium inhibitory agents, the delivery of such agents in the methods and compositions of the present disclosure can be significantly enhanced, the function of the agent being to reversibly inhibit the ciliary activity of mucosal cells, thereby providing a temporary, reversible increase in the residence time of the pharmaceutically active agent administered to the mucosa. When used in these aspects of the present disclosure, the cilium inhibitory factors described herein (whether their activity is direct or indirect) are all candidates for successful use as cilium inhibitory agents at appropriate doses (depending on concentration, duration, and mode of delivery), thereby producing a transient (i.e., reversible) reduction or cessation of mucociliary clearance at the site of administration of the mucosa to enhance the delivery of NMDP and other bioactive agents disclosed herein, without unacceptable side effects.

[0307] In more detailed aspects, specific cilia inhibitors can be used with NMDP and its salts and one or more other bioactive agents disclosed herein in a combined formulation or a coordinated administration scheme. Various bacterial cilia inhibitors isolated and characterized in the literature can be used in certain embodiments of the present disclosure. For example, cilia inhibitors from Pseudomonas aeruginosa include phenazine derivatives, pyo compounds (2-alkyl-4-hydroxyquinolines) and rhamnolipids (also known as hemolysins). In some embodiments, phenazine derivatives can also inhibit ciliary motility. In some embodiments, the composition may include rhamnolipids associated with changes in the ciliary membrane.

[0308] Surfactants and methods

[0309] In more detailed aspects of the present disclosure, one or more membrane permeation enhancers may be used in the mucosal delivery methods or formulations of the present disclosure to enhance mucosal delivery of NMDPs and other bioactive agents disclosed herein. The membrane permeation enhancers in this context may be selected from: (i) surfactants; (ii) bile salts; (iii) phospholipid additives, mixed micelles, liposomes or carriers; (iv) alcohols; (v) enamines; (vi) NO donor compounds; (vii) long-chain amphiphilic molecules; (viii) small hydrophobic permeation enhancers; (ix) sodium or salicylic acid derivatives; (x) acetoacetin; (xi) cyclodextrin or β-cyclodextrin derivatives; (xii) medium-chain fatty acids; (xiii) chelating agents; (xiv) amino acids or salts thereof; (xv) N-acetylamino acids or salts thereof; (xvi) enzymes that can degrade selected membrane components; (xvii) fatty acid synthesis inhibitors; (xviii) cholesterol synthesis inhibitors; or any combination of the membrane permeation enhancers described in (xix) (i)-(xviii).

[0310] Certain surfactants, also known as surfactants, can be readily incorporated into the mucosal delivery formulations and methods of the present disclosure as mucosal absorption enhancers. These agents, which can be co-administered or formulated in combination with NMDP and other bioactive agents disclosed herein, can be selected from a wide range of known surfactants. Surfactants are generally divided into three categories: (1) nonionic polyoxyethylene ethers, such as vitamin E TPGS and / or D-α-tocopheryl polyethylene glycol succinate; (2) bile salts, such as sodium glycocholate (SGC) and deoxycholate (DOC); and (3) fusidic acid and its derivatives, such as sodium taurodihydrofusidate (STDHF). The mechanism of action of these different classes of surfactants generally involves solubilizing the bioactive agent. For proteins and peptides that often form aggregates, the surface active properties of these absorption enhancers can allow interaction with the protein, thereby making it easier for smaller units (such as surfactant-coated monomers) to remain in solution. These monomers are presumably more easily transported than aggregates. Examples of other surfactants include L-α-lauroylphosphatidylcholine (DDPC), polysorbate 80, and polysorbate 20. Additional surfactants include polyethylene glycol, cetyl alcohol, polyvinyl pyrrolidone, polyvinyl alcohol, lanolin alcohol, sorbitan monooleate. In some embodiments, the surfactants of the present disclosure may be present in the pharmaceutical formulation alone or in any mixture or combination. In some embodiments, bile salts and certain fusidic acid derivatives have been reported to inhibit proteolytic degradation of proteins by nasal homogenates.

[0311] thickener

[0312] Thickeners or suspending agents can affect the release rate and / or absorption of the drug from the dosage form. Examples of materials that can be used as pharmaceutically acceptable thickeners include gelatin; methylcellulose (MC); hydroxypropyl methylcellulose (HPMC) and its derivatives; carboxymethylcellulose (CMC); cellulose; starch; hydroxyethyl starch; poloxamer; pluronic; sodium carboxymethylcellulose; sorbitol; gum arabic; povidone; carbomer (carbomer as used herein is a carbomer polymer, also known as type B carbomer homopolymer or 974PNF polymer); polycarbophil; chitosan; chitosan microspheres; alginate microspheres; glutamic acid chitosan; succinic acid resin; hyaluronic acid; ethylcellulose; maltodextrin DE; drum-dried corn starch (DDWM); degradable starch microspheres (DSM); deoxyglycocholate (GDC); hydroxyethyl cellulose (HEC); hydroxypropyl cellulose (HPC); microcrystalline cellulose (MCC); polymethacrylic acid and polyethylene glycol; sulfobutyl ether-β-cyclodextrin; cross-linked starch biospheres; sodium taurodihydrofusin (STDHF); N-trimethylchitosan chloride (TMC); degraded starch microspheres; succinic acid resin; chitosan nanoparticles; spray-dried crospovidone; spray-dried dextran microspheres; spray-dried microcrystalline cellulose; and cross-linked starch microspheres.

[0313] Carbomer thickeners useful herein also include, but are not limited to, the following: acrylic acid homopolymers, acrylic acid resins, acrylic acid polymers, acrylic acid resins, Acrysol A 1, Acrysol A 3, Acrysol A 5, Acrysol AC 5, Acrysol WS-24, Acrysol ase-75, Antiprex 461, Antiprex A, Arasorb 750, Arasorb S100F, Arolon, Aron, Aron A 10H, random poly(acrylic acid), CCRIS 3234, Carbomer 1342, Carbomer 910, Carbomer 1342, Carbomer 910, Carbomer 934, Carbomer 934P, Carbomer 940, Carbomer 941, Carbomer 960, Carbomer 961, Carbomer 971P, Carbomer 974P, Carbomer 980, Carbomer 981, Carboset 515, Carboset Resin 515, Carboxyvinyl Polymer, Carboxypolymethylene, Carpolene, Colloids 119 / 50, Cyguard 266, Dispex C40, Dow Latex 354, G-Cure, Good-rite K 37, Good-rite K 702, Good-rite K 732, Good-rite K-700, Good-rite K727, Good-rite WS 801, Haloflex 202, Haloflex 208, Joncryl 678, Junlon 110, Jurimer AC 10H, Jurimer AC 10P, NSC 106034, NSC 106035, NSC 106036, NSC106037, NSC 112122, NSC 112123, NSC 114472, NSC 165257, Nalfloc 636, Neocryl A-1038, OLD 01, P 11H, P 11H, P-11H, PA 11M, PAA-25, Pemulen TR-1, Pemulen TR-2, poly(acrylic acid), polyacrylate, polyacrylate elastomer, acrylic acid polymer, acrylic acid polymer cross-linked with pentaerythritol allyl ether, acrylic acid polymer cross-linked with pentaerythritol allyl ether, carboxyvinyl acrylic acid, Polytex 973, Primal ASE 60, acrylic acid polymer, R968, Racryl, Revacryl A 191, Rohagit SD 15, Sokalan PAS, Solidokoll N, Synthemul 90-588, TB1131, Tecpol, Texcryl, Versicol E 7, Versicol E15, Versicol E9, Versicol K 11, Versicol S25, Viscalex HV 30, Viscon 103, WS24, WS 801, XPA, etc. Other thickeners described in Ugwoke et al., Advances in Drug Delivery Reviews, 29:1656-57, 1998, are incorporated herein by reference. The pharmaceutical formulations disclosed herein may contain any one thickener or any combination or mixture of multiple thickeners.

[0314] Nitric oxide donor reagents and methods

[0315] In other related aspects of the present disclosure, nitric oxide (NO) donors are selected as membrane permeation enhancers to enhance mucosal delivery of one or more NMDPs and other bioactive agents disclosed herein. A variety of NO donors are known in the art and are used at effective concentrations in the methods and formulations of the present disclosure. Exemplary NO donors include, but are not limited to, nitroglycerin, sodium nitroprusside, NOC5 [3-(2-hydroxy-1-(methyl-ethyl)-2-nitrosohydrazino)-1-propylamine], NOC12 [N-ethyl-2-(1-ethyl-hydroxy-2-nitrosohydrazino)-ethylamine], SNAP [S-nitroso-N-acetyl-DL-penicillamine], NORI, and NOR4. In the methods and compositions of the present disclosure, an effective amount of a selected NO donor is co-administered or formulated in combination with one or more NMDPs and / or other bioactive agents disclosed herein to enter or cross the mucosal epithelium.

[0316] Agents that modulate epithelial junction structure and / or physiology

[0317] The present disclosure provides pharmaceutical compositions comprising one or more NMDPs and / or other bioactive agents, in combination with one or more mucosal delivery enhancers disclosed herein, formulated for mucosal delivery.

[0318] The permeabilizing agent typically reversibly enhances paracellular transport of mucosal epithelial cells by regulating the epithelial junction structure and / or physiology of the mucosal epithelial surface of the subject. This effect is generally related to the permeabilizing agent inhibiting the homotypic or heterotypic binding between the epithelial membrane adhesion proteins of adjacent epithelial cells. The target protein for blocking homotypic or heterotypic binding can be selected from various related tight junction adhesion molecules (JAMs), occludin or claudin proteins. Examples include antibodies, antibody fragments or single-chain antibodies that bind to the extracellular domains of these proteins.

[0319] In more detailed embodiments, the present disclosure provides permeabilizing peptides for enhancing paracellular transport in mucosal epithelial cells. The peptides generally function in the compositions and methods of the present disclosure by modulating epithelial junction structure and / or physiology in a mammalian subject. In certain embodiments, the peptides inhibit homotypic and / or heterotypic binding of epithelial membrane adhesion proteins selected from tight junction adhesion molecules (JAMs), occludin, or claudin.

[0320] One such agent that has been extensively studied is a bacterial toxin from Vibrio cholerae called "zonuloxin toxin" (ZOT). In these aspects of the present disclosure, ZOT is co-administered or formulated in combination with the active agent in the compositions and methods herein in an effective amount to significantly enhance the absorption of the active agent by reversibly increasing the permeability of the nasal mucosa without substantial side effects.

[0321] Vasodilators and methods

[0322] In some embodiments, the compositions and methods herein may include the administration of vasoactive compounds, more specifically vasodilators. These compounds are used in the present disclosure to modulate the structure and physiology of submucosal blood vessels, increase the transport rate of NMDPs and other bioactive agents into or across the mucosal epithelium and / or to specific target tissues or compartments (e.g., otic structures).

[0323] The vasodilators used in the present disclosure generally cause submucosal vasodilation by reducing cytoplasmic calcium concentration, increasing nitric oxide (NO), or inhibiting myosin light chain kinase. They are generally divided into nine categories: calcium antagonists, potassium channel openers, ACE inhibitors, angiotensin II receptor antagonists, α-adrenergic and imidazole receptor antagonists, β1-adrenergic agonists, phosphodiesterase inhibitors, eicosanoids, and NO donors.

[0324] In certain methods and compositions of the present disclosure, a selected vasodilator is co-administered (e.g., systemically or intranasally, simultaneously or in a combined effective time-dependent manner) or formulated in combination with one or more NMDPs in an amount effective to enhance mucosal absorption of the active agent to reach a target tissue or compartment (e.g., an otic structure) of a subject.

[0325] Selective transport enhancers and methods

[0326] The compositions and delivery methods disclosed herein may optionally incorporate selective transport enhancers to promote the transport of one or more bioactive agents. These transport enhancers can be used with one or more NMDP preparations disclosed herein in combination formulations or coordinated administration schemes to collaboratively enhance the delivery of one or more additional bioactive agents across mucosal transport barriers, thereby enhancing the mucosal delivery of the active agent to reach the target tissue or compartment (such as ear structure) of the subject. Alternatively, transport enhancers can be used in combination formulations or coordinated administration schemes to directly enhance the mucosal delivery of one or more NMDPs, whether or not enhancing the delivery of additional bioactive agents.

[0327] Exemplary selective transport enhancers used in this aspect of the present disclosure include, but are not limited to, glycosides, carbohydrate-containing molecules, and binders (such as lectin binders), which are known to specifically interact with epithelial transport barrier components. For example, specific "bioadhesive" ligands (including various plant and bacterial lectins) can bind to cell surface sugars through receptor-mediated interactions and are used as carriers or conjugated transport media in the present disclosure to enhance mucosal (e.g., intranasal) delivery of bioactive agents. Certain bioadhesive ligands in the present disclosure will mediate the transmission of biological signals to epithelial target cells, triggering the selective uptake of the adhesion ligand through specific cellular transport processes (endocytosis or transcytosis). Therefore, these transport media can be used as "carrier systems" to stimulate or guide the selective uptake of one or more NMDPs and other bioactive agents into and / or across mucosal epithelia. These and other selective transport enhancers can significantly enhance the mucosal delivery of large molecule biopharmaceuticals (especially peptides, proteins, oligonucleotides, and polynucleotide carriers) in the present disclosure. Lectins are plant proteins that can bind to specific sugars on the surface of eukaryotic cell glycoproteins and glycolipids. Concentrated solutions of lectins exhibit a "mucoadhesive" effect, and several studies have demonstrated rapid receptor-mediated endocytosis (RME) of lectins and lectin conjugates (such as concanavalin A conjugated to colloidal gold particles) across mucosal surfaces. Other studies have reported that the uptake mechanism of lectins can be used for intestinal drug targeting in vivo. In some of these studies, polystyrene nanoparticles (500 nm) covalently coupled to tomato lectin were reported to improve systemic uptake in rats after oral administration.

[0328] In addition to plant lectins, microbial adhesion and invasion factors provide a rich source of candidate adhesion / selective transport vectors for the mucosal delivery methods and compositions disclosed herein. The bacterial adhesion process requires two components: bacterial "adhesins" (adhesion or colonization factors) and receptors on the host cell surface. Bacteria that cause mucosal infections need to first penetrate the mucus layer and then attach to the epithelial surface. This attachment is usually mediated by bacterial pili or pili structures, although other cell surface components may also participate in this process. Adherent bacteria colonize the mucosal epithelium by proliferating and initiating a series of biochemical reactions in the target cell through signal transduction mechanisms (with or without the help of toxins). Related to these invasion mechanisms, a variety of biological adhesion proteins (such as invasins, intimins) produced by various bacteria and viruses are known. These enable microorganisms to attach extracellularly with remarkable selectivity for host species and even specific target tissues. The signals transmitted by such receptor-ligand interactions trigger the transport of intact live microorganisms into and ultimately across epithelial cells through endocytosis and transcytosis. According to the teachings herein, such natural phenomena can be exploited (eg, by complexing bioactive agents such as NMDPs with adhesins) to enhance the delivery of bioactive compounds into or across mucosal epithelia and / or to other designated drug target sites.

[0329] Various bacterial and plant toxins that bind to epithelial surfaces in a specific lectin-like manner can also be used in the methods and compositions of the present disclosure. For example, diphtheria toxin (DT) rapidly enters host cells via RME. Similarly, the B subunit of the Escherichia coli heat-labile toxin binds to the brush border of intestinal epithelial cells in a highly specific lectin-like manner. Uptake and transcytosis of this toxin to the basolateral side of intestinal cells have been reported to occur both in vivo and in vitro. Other studies have expressed the transmembrane domain of diphtheria toxin in Escherichia coli as a maltose-binding fusion protein and chemically coupled it to high-molecular-weight poly-L-lysine. The resulting complex was successfully used to mediate the internalization of a reporter gene in vitro. In addition to these examples, Staphylococcus aureus produces a group of proteins (such as Staphylococcus aureus enterotoxin A (SEA), SEB, and toxic shock syndrome toxin 1 (TSST-1)) that act as both superantigens and toxins. Studies on these proteins have reported that SEB and TSST-1 promote transcytosis in a dose-dependent manner in Caco-2 cells.

[0330] Viral hemagglutinins constitute another class of transporters that promote mucosal delivery of bioactive agents in the methods and compositions of the present disclosure. The initial step in many viral infections is the binding of surface proteins (hemagglutinins) to mucosal cells. These binding proteins have been identified in most viruses, including rotavirus, varicella zoster virus, Semliki Forest virus, adenovirus, potato leafroll virus, and reovirus. These and other exemplary viral hemagglutinins can be used with one or more NMDPs disclosed herein in combination formulations (such as mixtures or conjugate formulations) or coordinated administration schemes to synergistically enhance the mucosal delivery of one or more additional bioactive agents. Alternatively, viral hemagglutinins can be used in combination formulations or coordinated administration schemes to directly enhance the mucosal delivery of one or more NMDPs, whether or not enhancing the delivery of additional bioactive agents.

[0331] A variety of endogenous selective transport mediators can also be used in the present disclosure. Mammalian cells have developed a variety of mechanisms to promote the internalization of specific substrates and target them to specific compartments. In general, these membrane deformation processes are called "endocytosis", including phagocytosis, pinocytosis, receptor-mediated endocytosis (clathrin-mediated RME) and caveolae (non-clathrin-mediated RME). As the name suggests, RME is a highly specific cell biological process by which various ligands bind to cell surface receptors and are subsequently internalized and transported within the cell. In many cells, endocytosis is very active and the entire membrane surface can be internalized and replaced in less than half an hour. Based on its orientation in the cell membrane, two types of receptors have been proposed: the amino terminus of type I receptors is located on the extracellular side of the membrane, while the same protein tail of type II receptors is located in the intracellular environment.

[0332] Other embodiments of the present disclosure utilize transferrin as an RME carrier or stimulator for mucosal delivery of bioactive agents. Transferrin is an 80kDa iron transport glycoprotein that is efficiently taken up into cells through RME. Transferrin receptors are present on the surface of most proliferating cells, and the number on erythroblasts and various tumors is increased. It is reported that in the presence of the fungal metabolite brefeldin A (BFA), the transcytosis of transferrin (Tf) and transferrin conjugates is enhanced. In other studies, it is reported that BFA treatment can rapidly increase the apical endocytosis of ricin and HRP in MDCK cells. Therefore, BFA and other agents that stimulate receptor-mediated transport can be used as combined formulations (such as conjugation) and / or co-administered agents in the methods of the present disclosure to enhance receptor-mediated transport of bioactive agents, such as NMDP.

[0333] Polymer delivery vehicles and methods

[0334] In certain aspects of the present disclosure, NMDP, other bioactive agents disclosed herein, and delivery enhancers described herein can be incorporated into a formulation for mucosal (e.g., intranasal) administration, either alone or in combination, comprising a biocompatible polymer used as a carrier or matrix. Such polymer carriers include polymer forms such as polymer powders, matrices, or microparticle delivery vehicles. The polymer can be derived from plants, animals, or synthetics. The polymer is typically a cross-linked structure. In addition, in these delivery systems, the NMDP can be functionally modified so that it can be covalently bound to the polymer and inseparable from the polymer. In other embodiments, the polymer is chemically modified with enzyme inhibitors or other agents that can degrade or inactivate the bioactive agent and / or delivery enhancer. In certain formulations, the polymer is a partially or completely water-insoluble but water-swellable polymer, such as a hydrogel. In some embodiments, the polymer is water-interactive and / or hydrophilic in nature.

[0335] To prolong the biological activity of NMDP and other bioactive agents disclosed herein, as well as optional delivery enhancers, these agents can be incorporated into a polymer matrix, such as a polyorthoester, polyanhydride, or polyester. In some embodiments, this can result in sustained activity and release of the active agent, as determined, for example, by degradation of the polymer matrix. The absorption-enhancing polymers contemplated for use in the present disclosure may include derivatives and chemically or physically modified forms of the above-mentioned types of polymers, as well as other natural or synthetic polymers, gums, resins, and other agents, and blends of these materials with each other or with other polymers. In certain aspects of the formulations disclosed herein, synthetic polymers such as nylon, acrylic, and other generally hydrophobic polymers can be sufficiently modified by reaction to become water-swellable and / or form stable gels in aqueous media. The absorption-enhancing polymers disclosed herein may include homopolymers and copolymers based on various vinyl monomer combinations: acrylic acid and methacrylic acid, acrylamide, methacrylamide, hydroxyethyl acrylate or hydroxyethyl methacrylate, vinyl pyrrolidone, as well as polyvinyl alcohol and its copolymers and terpolymers, polyvinyl acetate and its copolymers with the above-mentioned monomers and 2-acrylamido-2-methylpropanesulfonic acid. Copolymer and terpolymer.In some embodiments, the copolymer of above-mentioned monomer and copolymerizable functional monomer (such as acryloyl or methacrylamide acrylate or methacrylate) can be used.Other absorption promoting polymers expected in the present disclosure are the polymers classified as dextran, dextrin, and the material classified as natural gum and resin, or the material classified as natural polymer, such as processed collagen, chitin, chitosan, pullulan, extracellular polysaccharide Zooglan, alginate and modified alginate, such as " Kelcoloid " (a kind of polypropylene glycol modified alginate)), gellan gum such as " Kelocogel ", xanthan gum such as " Keltrol ", estastin, alpha-hydroxybutyric acid and copolymer thereof, hyaluronic acid and derivative thereof, polylactic acid and glycolic acid.

[0336] In some embodiments, the polymers contemplated in the compositions and methods of the present disclosure are ethylenically unsaturated carboxylic acids containing at least one activated carbon-carbon olefinic double bond and at least one carboxyl group; that is, a functional group readily converted to an acid containing an olefinic double bond, which is readily functional in polymerization due to its presence in the monomer molecule in an α-β position relative to the carboxyl group or as part of a terminal methylene group. Such olefinically unsaturated acids include acrylic acids such as acrylic acid itself, α-cyanoacrylic acid, β-methacrylic acid (crotonic acid), α-phenylacrylic acid, β-acryloyloxypropionic acid, cinnamic acid, p-chlorocinnamic acid, 1-carboxy-4-phenylbutadiene-1,3, itaconic acid, citraconic acid, mesaconic acid, glutaric acid, aconitic acid, maleic acid, fumaric acid, and ethylene tricarboxylate.

[0337] In some embodiments contemplated in the compositions and methods herein, absorption enhancers may include methyl acrylate, ethyl acrylate, propyl acrylate, isopropyl acrylate, butyl acrylate, isobutyl acrylate, methyl methacrylate, ethyl methacrylate, ethyl methacrylate, octyl acrylate, heptyl acrylate, octyl methacrylate, isopropyl methacrylate, 2-ethylhexyl methacrylate, nonyl acrylate, hexyl acrylate, n-hexyl methacrylate, and the like. Higher alkyl acrylates include decyl acrylate, isodecyl methacrylate, lauryl acrylate, stearyl acrylate, behenyl acrylate, and their methacrylate versions. Mixtures of two or more long-chain acrylates can be successfully polymerized with one of the carboxylic acid monomers. Other comonomers include olefins, including α-olefins, vinyl ethers, vinyl esters, and mixtures thereof.

[0338] Other vinylidene monomers, including acrylic nitrile, can also be used as absorption enhancers in the methods and compositions of the present disclosure to enhance the delivery and absorption of one or more NMDPs and other bioactive agents herein, including enhancing the delivery of the active agent to a target tissue or compartment of a subject (e.g., an ear structure). In some embodiments, the α,β-ethylenically unsaturated nitrile can be a monoethylenically unsaturated nitrile having 3 to 10 carbon atoms, such as acrylonitrile, methacrylonitrile, etc. Acrylamides containing 3 to 35 carbon atoms can be used, including monoethylenically unsaturated amides. Representative amides include acrylamide, methacrylamide, N-tert-butylacrylamide, N-cyclohexylacrylamide, higher alkyl amides (wherein the alkyl group on the nitrogen contains from 8 to 32 carbon atoms), acrylamides, including N-alkanolamides of α,β-ethylenically unsaturated carboxylic acids (including those having from 4 to 10 carbon atoms), such as N-methylolacrylamide, N-propanolacrylamide, N-methylolmethacrylamide, N-methylolmaleimide, N-methylolmaleamate, N-methylolp-vinylbenzamide, and the like. In other embodiments, the absorption-enhancing material is an alpha-olefin containing from 2 to 18 carbon atoms (more preferably from 2 to 8 carbon atoms); a diene containing from 4 to 10 carbon atoms; vinyl esters and allyl esters (such as vinyl acetate); vinyl aromatic hydrocarbons (such as styrene, methylstyrene, and chlorostyrene); vinyl and allyl ethers and ketones (such as vinyl methyl ether and methyl vinyl ketone); chloroacrylates; cyanoalkyl acrylates (such as alpha-cyanomethacrylate, and alpha-, beta-, and gamma-cyanopropyl acrylate); Ester); alkoxy acrylate (such as methoxyethyl acrylate); haloacrylate (such as chloroethyl acrylate); vinyl halide and vinyl chloride, vinylidene chloride, etc.; divinyl compounds, diacrylates and other multifunctional monomers (such as divinyl ether, diethylene glycol diacrylate, ethylene glycol dimethacrylate, methylenebisacrylamide, allyl pentaerythritol, etc.); and bis (β-haloalkyl) alkenyl phosphonates (such as bis (β-chloroethyl) vinyl phosphonate, etc.), as known to those skilled in the art. Copolymers in which carboxyl-containing monomers are minor components and other vinylidene monomers are present as major components can be easily prepared according to the methods disclosed herein.

[0339] In some embodiments, the absorption enhancers contemplated in the methods and compositions herein may be composed of synthetic copolymers of acrylic and methacrylic acid, acrylamide, methacrylamide, hydroxyethyl acrylate (HEA) or hydroxyethyl methacrylate (HEMA), and water-interactive and swellable vinyl pyrrolidone. Other very useful hydrogel polymers are swellable but insoluble versions of poly(vinyl pyrrolidone) starch, carboxymethyl cellulose, and polyvinyl alcohol. In some embodiments, polymer hydrogel materials contemplated in the present disclosure include (poly)hydroxyalkyl (meth)acrylates: anionic and cationic hydrogels: poly(electrolyte) complexes: poly(vinyl alcohol) with low acetate residues: swellable mixtures of cross-linked agar and cross-linked carboxymethyl cellulose: swellable compositions comprising methyl cellulose mixed with a small amount of cross-linked agar: water-swellable copolymers produced from finely divided copolymer dispersions of maleic anhydride with styrene, ethylene, propylene, or isobutylene: water-swellable polymers of N-vinyl lactam: swellable sodium salts of carboxymethyl cellulose, etc.

[0340] In some embodiments, polymers contemplated herein for mucosal delivery of bioactive agents (including NMDPs) include pectin; polysaccharides (such as agar, gum arabic, gum karaya, gum tragacanth, alginates, and guar gum and cross-linked versions thereof); acrylic acid polymers, copolymers, and salt derivatives, polyacrylamides; water-swellable indene maleic anhydride polymers; starch graft copolymers; acrylate polymers and copolymers having a water absorption capacity of about 2 to 400 times their original weight; polyglucan diesters; a mixture of cross-linked poly(vinyl alcohol) and poly(N-vinyl-2-pyrrolidone); polyoxybutylene-polyethylene block copolymer gels; carob bean gum; polyester gels; polyurea gels; polyether gels; polyamide gels; polyimide gels; polypeptide gels; polyamino acid gels; polycellulose gels; cross-linked indene-maleic anhydride acrylate polymers; and polysaccharides. In some embodiments, the hydrogel polymers contemplated in the present disclosure are cross-linked to effectively contain the bioactive agent.

[0341] The cross-linked network can be formed by free radical copolymerization of unsaturated monomers. Polymer hydrogels can also be formed by reacting functional groups on the polymer (such as alcohols, acids, amines) with glyoxal, formaldehyde or glutaraldehyde, dianhydride, etc. to crosslink preformed polymers. The polymer can also be cross-linked with any polyene (such as decadiene or trivinylcyclohexane); acrylamide (such as N,N-methylenebis(acrylamide)); multifunctional acrylate (such as trimethylolpropane triacrylate); or a multifunctional vinylidene monomer containing at least 2 terminal CH2< groups (including, for example, divinylbenzene, divinylnaphthalene, allyl acrylate, etc.). In certain embodiments, the cross-linking monomer for preparing the copolymer is a polyalkenyl polyether having one or more alkenyl ether groups per molecule, which may optionally have an alkenyl group, wherein there is an olefinic double bond connected to the terminal methylene group (for example, prepared by etherification of a polyol containing at least 2 carbon atoms and at least 2 hydroxyl groups). Other crosslinking monomers include, for example, diallyl esters, dimethyl allyl ethers, allyl or methallyl acrylates and acrylamides, tetravinylsilane, polyalkenyl methane, diacrylates and dimethacrylates, divinyl compounds such as divinylbenzene, polyalkenyl phosphates, diallyloxy compounds, and phosphites, and the like.

[0342] In other aspects of the present disclosure, mucosal delivery of NMDP and other bioactive agents disclosed herein can be enhanced by retaining the active agent in a slow-release or enzyme or physiologically protective carrier or vehicle (e.g., a hydrogel that protects the active agent from the action of degradative enzymes). In certain embodiments, the active agent is chemically bound to the carrier or vehicle, and additional reagents (such as enzyme inhibitors, cytokines, etc.) can also be mixed or bound to the carrier or vehicle. The active agent can alternatively be immobilized by being fully physically embedded in a carrier or vehicle (e.g., a polymer matrix).

[0343] The polymers (e.g., hydrogels) disclosed herein may incorporate functional linking agents, such as glycosides chemically incorporated into the polymer, to enhance the intranasal bioavailability of the active agent formulated therewith. Examples of such glycosides include glucosides, fructosides, galactosides, arabinosides, mannosides, and their alkyl-substituted derivatives, and natural glycosides such as arbutin, phlorizin, amygdalin, digitonin, saponins, and indigosides. Typical glycosides can be incorporated into the polymer in several ways. For example, the hydrogen of the hydroxyl group of a glycoside or other similar carbohydrate can be replaced by an alkyl group from the hydrogel polymer to form an ether. In addition, the hydroxyl group of the glycoside can react to esterify the carboxyl group of the polymer hydrogel to form a polymer ester in situ. Another method is to use the condensation of acetylglucose bromide and cholester-5-en-3β-ol on a maleic acid copolymer. N-substituted polyacrylamides can be synthesized by reacting activated polymers with ω-aminoalkyl glycosides: (1) (carbohydrate-spacer) (n)-polyacrylamide, a "pseudopolysaccharide"; (2) (carbohydrate-spacer) (n)-phosphatidylethanolamine (m)-polyacrylamide, a neoglycolipid, a derivative of phosphatidylethanolamine; and (3) (carbohydrate-spacer) (n)-biotin (m)-polyacrylamide. These biotinylated derivatives can be attached to lectins on mucosal surfaces to promote the absorption of bioactive agents (such as polymer-encapsulated NMDP).

[0344] Bioadhesive delivery vehicles and methods

[0345] In some embodiments, the combination formulations and / or coordinated administration methods herein incorporate an effective amount of a non-toxic bioadhesive as an adjunct compound or carrier to enhance mucosal delivery of one or more bioactive agents. Bioadhesives in this context exhibit general or specific adhesion to one or more components or surfaces of the target mucosa. This enhancement of epithelial permeation generally allows for effective transmucosal delivery of macromolecules, for example, to the basal portion of the nasal epithelium or into adjacent extracellular compartments or plasma or central nervous system tissues or fluids. In some embodiments, the bioadhesives disclosed herein can be used in the combination formulations and coordinated administration methods of the present disclosure, which optionally incorporate an effective amount and form of bioadhesive to prolong the persistence of one or more NMDPs and other bioactive agents or otherwise increase their mucosal absorption. Bioadhesives can be co-administered as adjunct compounds or incorporated into the combination formulations of the present disclosure as additives. In certain embodiments, the bioadhesive acts as a "drug glue," while in other embodiments, adjunctive delivery or combined formulations of bioadhesives are used to enhance contact of bioactive agents with the nasal mucosa, in some embodiments by promoting specific receptor-ligand interactions with epithelial cell "receptors," and in other embodiments by increasing epithelial permeability to significantly increase the drug concentration gradient measured at the target site (e.g., liver, plasma, or central nervous system tissue or fluid). Additional bioadhesives in the present disclosure act as enzyme (e.g., protease) inhibitors to enhance the stability of mucosally administered biotherapeutics delivered in a coordinated or combined formulation with a bioadhesive.

[0346] By determining the ability of various bioadhesive polymers to retain and release NMDPs, as well as their ability to interact with mucosal surfaces after incorporation of an active agent therein, their potential as mucosal (e.g., intranasal) delivery platforms in the methods and compositions of the present disclosure can be readily assessed. Furthermore, the biocompatibility of the selected polymer with the tissue at the mucosal application site is determined using known methods. When the target mucosa is covered with mucus (i.e., in the absence of mucolytic or mucus-clearing treatments), it can serve as a connecting link to the underlying mucosal epithelium. Therefore, the term "bioadhesive," as used herein, also encompasses mucoadhesive compounds used to enhance mucosal delivery of bioactive agents in the present disclosure. However, adhesive contact with mucosal tissue mediated by adhesion to a mucus gel layer can be limited by incomplete or transient adhesion between the mucus layer and the underlying tissue, particularly on nasal surfaces where rapid mucus clearance occurs. In this regard, mucin glycoproteins are continuously secreted and form viscoelastic gels immediately upon release from cells or glands. However, the luminal surface of the adhesive gel layer is continuously eroded by mechanical, enzymatic, and / or ciliary action. In cases where these activities are more prominent or longer adhesion times are desired, the coordinated administration methods and combined formulation methods herein may further incorporate mucolytic and / or ciliary inhibitory methods or agents as disclosed above.

[0347] In general, the mucus-adhesive polymers used in this disclosure are natural or synthetic macromolecules that adhere to the surface of moist mucosal tissues through complex but non-specific mechanisms. In addition to these mucus-adhesive polymers, this disclosure also describes methods and compositions of bioadhesives that adhere directly to cell surfaces rather than mucus through specific (including receptor-mediated) interactions. One example of a bioadhesive that acts in this specific manner is the group of compounds known as lectins.

[0348] In certain aspects of the present disclosure, bioadhesive materials for enhancing intranasal delivery of bioactive agents comprise a matrix of a hydrophilic (e.g., water-soluble or swellable) polymer or polymer mixture that adheres to wet mucosal surfaces. These adhesives can be formulated as ointments, hydrogels (see above), films, and other forms of administration. Typically, these adhesives are mixed with bioactive agents to achieve slow release or local delivery of the active agent. Some formulations contain additional ingredients to promote penetration of the active agent across the nasal mucosa, for example, into the ear canal of an individual.

[0349] Various natural and synthetic polymers exhibit significant binding to mucus and / or mucosal epithelial surfaces under physiological conditions. The strength of this interaction can be easily measured by mechanical peeling or shear tests. When applied to a moist mucosal surface, many dry materials will spontaneously adhere, at least slightly. After this initial contact, some hydrophilic materials begin to attract water through adsorption, swelling, or capillary forces. If this water is absorbed from the underlying matrix or from the polymer-tissue interface, the adhesion may be sufficient to achieve the goal of enhancing mucosal absorption of the bioactive agent. This "adhesion through hydration" can be quite strong, but formulations employing this mechanism must take into account that the drug will continue to swell after conversion to hydrated mucus (e.g., certain cellulose derivatives), which are generally not adhesive in the pre-hydrated state. In some embodiments, such materials for mucosal administration in the present disclosure are effective when the bioadhesive drug delivery system is applied in the form of dry polymer powders, microspheres, or film-type delivery.

[0350] Acrylic acid-based hydrogels are well-suited for bioadhesion due to their flexibility and non-abrasive properties in a partially swollen state, which reduces wear that can damage contacting tissue. In some embodiments, the methods and compositions of the present disclosure optionally include the use of a carrier, such as a polymeric delivery vehicle, which functions in part to protect the bioactive agent from proteolytic degradation while providing enhanced penetration of the peptide or protein into or through the nasal mucosa, such as the mucoadhesive poly(acrylic acid) derivative polycarbophil.

[0351] Other mucoadhesive polymers disclosed herein, such as chitosan, have been reported to enhance the permeability of certain mucosal epithelia even when administered in aqueous solution or gel form. Another mucoadhesive polymer reported to directly affect epithelial permeability is hyaluronic acid and its ester derivatives. Chitosan is the N-deacetylated product of chitin, a naturally occurring polymer that has been widely used to prepare microspheres for oral and intranasal formulations. In one aspect of the disclosure, O-methylisourea is used to convert chitosan amines to their guanidino moieties. For example, guanidino compounds are prepared by reacting isotactic solutions of chitosan and O-methylisourea at a pH above 8.0.

[0352] In some embodiments, the bioadhesive is a lectin. Lectins are (glyco)proteins of non-immune origin that bind to polysaccharides or glycoconjugates. Several plant lectins have been studied as possible drug absorption enhancers. One plant lectin, Phaseolus vulgaris hemagglutinin (PHA), exhibits a high oral bioavailability of over 10% after feeding to rats. Tomato (Lycopersicon esculeutum) lectin (TL) appears to be safe for various routes of administration.

[0353] Microemulsion delivery method

[0354] In some embodiments, the NMDP intranasal delivery formulation may comprise a microemulsion system stabilized by a nonionic surfactant (such as CremophorRH 40 or Labrasol) and may comprise various oils, including isopropyl myristate, LabrafilM1944CS or Maisine 35-1. The formulation may comprise 8% Labrafil M 1944CS, 30% Cremophor RH 40 / ethanol (3:1) and water, with an NMDP solubility of up to 6.4 mg / ml, a droplet size of 30.3+ / -5.3 nm, and no ciliary toxicity. Following a single intranasal administration of this formulation at a dose of 2 mg / kg, plasma concentrations may reach a peak in 1 hour (such as C max ), the absolute bioavailability is about 32%.

[0355] Liposome and micelle delivery vehicles

[0356] The co-administration methods and combined formulations disclosed herein are optionally incorporated with effective lipid or fatty acid-based carriers, processing agents, or delivery vehicles to provide improved formulations for mucosal delivery of NMDP and other bioactive agents. For example, a variety of formulations and methods for mucosal delivery are provided, comprising one or more of these active agents, in addition to peptides or proteins, mixed or encapsulated with liposomes, mixed micelle carriers, or emulsions, or co-administered therewith, to enhance the chemical and physical stability of NMDP and other active agents herein during mucosal delivery and increase their half-life.

[0357] In some embodiments, the delivery system for bioactive agents comprises small lipid vesicles called liposomes. They can be made of natural, biodegradable, non-toxic and non-immunogenic lipid molecules and can effectively embed or bind drug molecules into or on their membranes.

[0358] In other embodiments, nimodipine and other active agents described herein can be combined with the use of polymers and liposomes to combine the advantageous properties of the two carriers, such as being embedded in the natural polymer fibrin and controlled release by using covalent cross-linking and adding antifibrinolytic agents to the fibrin polymer. In other embodiments, liposomes may comprise cationic lipids, long-chain and medium-chain fatty acids, and surfactant mixed micelles with fatty acids, long-chain fatty acids, fusion lipids, unsaturated fatty acids and monoglycerides (such as oleic acid, linoleic acid, linoleic acid, monoolein glyceride, etc.), medium-chain fatty acids (C6 to C12) and monoglycerides, sodium salts of medium-chain fatty acids (C6 to 12) and carriers to enhance mucosal delivery of NMDP and other bioactive agents disclosed herein. Fatty acids can be used in a soluble form of a sodium salt, or by adding a non-toxic surfactant (such as polyoxyethylene hydrogenated castor oil, sodium taurocholate, etc.). Fatty acids and mixed micelle formulations contemplated in the present disclosure include, but are not limited to, sodium caprylate (C8), sodium caprate (C10), sodium laurate (C12), or sodium oleate (C18), optionally in combination with bile salts such as glycocholate and taurocholate.

[0359] PEGylation

[0360] In alternative embodiments, NMDP salts are conjugated to polyalkylene oxide polymers, particularly polyethylene glycol (PEG), with other bioactive peptides and proteins. See U.S. Patent No. 4,179,337. PEG polymers contemplated in the present disclosure include SC-PEG with molecular weights of 2000, 5000, 10000, 12000, and 20000; U-PEG-10000; NHS-PEG-3400-biotin; T-PEG-5000; T-PEG-12000; and TPC-PEG-5000.

[0361] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0362] Example

[0363] The following examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0364] Example 1: NMDP intranasal formulation

[0365] In the present embodiment, NMDP is dissolved in the solution containing carrier, citrate buffer, benzalkonium chloride and water, and the NMDP solution that concentration is about 50mg / mL is obtained by ultrasonic treatment. Subsequently, 50 microlitres of said compositions are applied to each nasal cavity of experimenter using intranasal drug delivery device. Blood samples are collected at 0, 2, 5, 10, 15, 30 and 60 minutes, and NMDP concentration is measured by high performance liquid chromatography (HPLC). Subsequently, the pharmacokinetics of intranasal administration of NMDP can be compared with the pharmacokinetics of intravenous administration of NMDP. The pharmacokinetics of intranasal administration of NMDP is suitable with the pharmacokinetics of intravenous administration of NMDP.

[0366] Example 2: NMDP intranasal formulation containing surfactant

[0367] In this embodiment, NMDP is dissolved in a solution containing one or more natural or synthetic carriers (content of about 30% to about 95% (w / w)), a non-aqueous solvent (about 10% to about 70% (w / w)) and a surfactant, and an NMDP solution having a concentration of about 50 mg / mL is obtained by ultrasonic treatment. Subsequently, 50 microliters of the composition is applied to each nasal cavity of the subject using an intranasal drug delivery device. Blood samples are collected at 0, 2, 5, 10, 15, 30 and 60 minutes, and the NMDP concentration is determined by high performance liquid chromatography (HPLC). The pharmacokinetics of intranasally administered NMDP can then be compared with the pharmacokinetics of intravenously administered NMDP. The pharmacokinetics of intranasally administered NMDP are comparable to those of intravenously administered NMDP.

[0368] Example 3: Surfactant-free NMDP intranasal formulation

[0369] In this embodiment, NMDP is dissolved in a solution comprising about 60% to about 80% (w / w) mPEG350, about 10% to about 30% (w / w) PEG400, about 5% to about 15% (w / w) water and not containing a nonionic surfactant. Subsequently, 50 microliters of a composition comprising about 100 mg / mL NMDP is applied to each nasal cavity of the subject using an intranasal drug delivery device. Blood samples are collected at 0, 2, 5, 10, 15, 30, and 60 minutes, and the NMDP concentration is determined by high performance liquid chromatography (HPLC). The pharmacokinetics of intranasal administration of NMDP can then be compared with the pharmacokinetics of intravenous administration of NMDP. The pharmacokinetics of intranasal administration of NMDP is comparable to the pharmacokinetics of intravenous administration of NMDP.

[0370] Example 4: Intranasal NMDP Formulation without Surfactant but Containing Ethanol

[0371] In the present embodiment, NMDP is dissolved in a solution comprising about 60% to about 80% (w / w) mPEG350, about 10% to about 30% (w / w) PEG400, about 5% to about 15% (w / w) water, about 5% to about 15% ethanol and not containing a nonionic surfactant. Subsequently, 50 microliters of a composition comprising about 100 mg / mL NMDP is applied to each nasal cavity of the subject using an intranasal drug delivery device. Blood samples are collected at 0, 2, 5, 10, 15, 30 and 60 minutes, and the NMDP concentration is determined by high performance liquid chromatography (HPLC). The pharmacokinetics of intranasal administration of NMDP can then be compared with the pharmacokinetics of intravenous administration of NMDP. The pharmacokinetics of intranasal administration of NMDP is comparable to the pharmacokinetics of intravenous administration of NMDP.

[0372] Example 5: Preparation and intranasal preparation of NMDP

[0373] NMDP can be formulated into a nasal composition and administered to a patient as a drug for the treatment of one of the indications described herein. In short, NMDP, a buffer, benzalkonium chloride, and optionally other ingredients (such as sodium chloride or other osmotic pressure regulators, sorbitol or other sweeteners, flavorings, etc.) can be formulated into a solution having a volume less than the target final volume. The ingredients are then mixed until completely dissolved. If necessary, the pH can be adjusted by adding a suitable acid or base (such as HCl, NaOH, or a complementary acid or base to the buffer). After the desired pH is reached, water can be added to bring the solution to a specified volume. The resulting solution can be dispensed into suitable containers for transport and distribution. In some embodiments, the suitable container includes a nasal pump. In other embodiments, the suitable container can be a vial, such as an amber glass bottle, which can be a glass ampoule, a glass bottle topped with an inert rubber septum and a crimped cap, or other suitable pharmaceutical vial.

[0374] Example 6: Treating Tinnitus with NMDP

[0375] A subject suffering from tinnitus is treated by intranasal administration of about 1-10 mg / kg of NMDP. The subject is treated using the intranasal delivery formulation of F1 or F2 in Example 15. max The duration was approximately 20 minutes. Subjects noted a reduction in tinnitus or other tinnitus symptoms following intranasal administration. Subjects were re-administered as needed to treat tinnitus symptoms, approximately every 2-4 hours. Lower doses of NMDP were administered to subjects using the intranasal delivery formulation compared to oral administration.

[0376] Example 7: Intranasal Administration of NDMP

[0377] Six volunteers with tinnitus were treated with approximately 1-10 mg / kg of NMDP intranasally. A nasal spray was prepared as described in Example 5, with 50 μL administered to each nostril every two to four hours. A 100 μL dose delivered approximately 20 mg of NMDP. Blood samples were collected at 0, 2, 5, 10, 15, 30, and 60 minutes, and NMDP concentrations were determined by high-performance liquid chromatography (HPLC). max The time is about 1 hour. The pharmacokinetics of intranasal NMDP can then be compared with the pharmacokinetics of intravenous NMDP. The pharmacokinetics of intranasal NMDP are comparable to those of intravenous NMDP.

[0378] Example 8: Intranasal vesicle composition

[0379] In this example, approximately 2500 mg of NMDP was dissolved in a solution containing a phospholipid, one or more C2-C4 alcohols, and water to produce a solution containing approximately 200 mg / mL of NMDP. This composition contained vesicles ranging in size from 50 nm to several microns, more specifically up to 5 μm, which exhibit promising properties for enhanced nasal absorption. The vesicles were then observed using transmission electron microscopy (TEM) and scanning electron microscopy. TEM analysis was performed using an electron microscope operating at an accelerating voltage of 100 kV.

[0380] Then, 50 μL of the composition was administered to each nasal cavity of the subject using an intranasal administration device. Blood samples were collected at 0, 2, 5, 10, 15, 30, and 60 minutes, and NMDP concentrations were determined by high performance liquid chromatography (HPLC). The pharmacokinetics of intranasal administration of NMDP were then compared with those of intravenous administration of NMDP. The pharmacokinetics of intranasal administration of NMDP were comparable to those of intravenous administration of NMDP.

[0381] Example 9: NMDP dry powder formulation

[0382] Approximately 2500 mg of NMDP was dissolved in a mixture of acetone (12 g) and ethanol (12 g) with stirring at 300 rpm. A suitably sized magnetic bar was placed in a receiver, and lactose monohydrate (2.3 g) was added, with the stirring rate set to 150 rpm. The clear, homogeneous solution of the active agent (sumatriptan succinate) was spray-dried at an inlet air temperature of 60°C and an outlet temperature of 55°C to obtain a dry active agent powder, which was then blended in situ with lactose monohydrate in the receiver. Stirring was maintained throughout the process. The actual weight of NMDP as the active agent in the resulting sumatriptan / lactose composition was approximately 15% w / w. This composition was then mixed with additional lactose to achieve the desired active agent (NMDP) concentration of 10%.

[0383] Example 10: Mucoadhesive Preparation

[0384] A 20 g batch of mucoadhesive gel formulation containing 2.0% NMDP was prepared by suspending 20.0 mg of carbomer 934P and 1.80 g of poloxamer 188 (BASF) in 5.00 g of TRIS-HCl buffer (0.1 M) and stirring overnight at 4°C to ensure complete dissolution. Additional ingredients, including hydroxypropyl methylcellulose (100.0 mg), methylparaben (10 mg), and additional TRIS-HCl buffer (0.1 M) (2.87 g), were added and stirring continued until complete dissolution. In addition to NMDP, tacrolimus (100 g) was added and mixed while maintaining activity. The mixture was maintained below room temperature until use.

[0385] Example 11: Microemulsion Formulation

[0386] NMDP was dissolved in a microemulsion stabilized by nonionic surfactants (Cremophor RH 40 and Labrasol) at a concentration of approximately 50 mg / mL. The system contained an oil phase consisting of isopropyl myristate, Labrafil M 1944CS, and Maisine 35-1. This formulation, comprising 8% Labrafil M 1944CS, 30% Cremophor RH 40 / ethanol (3:1), and water, achieved a maximum solubility of 6.4 mg / mL for NMDP, a droplet size of 30.3 ± 5.3 nm, and was non-ciliary toxic.

[0387] Subsequently, 50 μL of the composition was administered to each nasal cavity of the subject using an intranasal administration device. Blood samples were collected at 0, 2, 5, 10, 15, 30, and 60 minutes, and NMDP concentrations were determined by high performance liquid chromatography (HPLC). The pharmacokinetics of intranasal NMDP were then compared with those of intravenous NMDP. Following a single intranasal administration of the formulation at a dose of 2 mg / kg, plasma concentrations peaked at 1 hour, with an absolute bioavailability of approximately 32%.

[0388] Example 12: Treatment of induced tinnitus in mice

[0389] In this example, mice with noise-induced tinnitus were treated with an intranasal delivery formulation of NMDP (such as the formulation of Example 1).

[0390] Use Figure 3The Sound-Based Avoidance Detection (SBAD) assay, described here, detects tinnitus and tests animals' responses to different pharmacological doses of NMDP. Using a two-compartment shuttle box, mice were trained to shuttle back and forth in response to a sound cue during "Go" trials and remain motionless in the absence of a sound during "No-Go" trials for 15 days. Before testing began, animals were acclimated to the shuttle box for 5 minutes, with 100 trials randomly assigned daily for approximately 30-40 minutes. The sound cue consisted of either white noise or narrowband noise at 5, 10, 16, 20, and 32 kHz, with a randomized intensity of 75, 80, or 85 dB. To reinforce training, mice were given an electric shock if they failed to shuttle back and forth during "Go" trials or moved between compartments in the absence of a sound during "No-Go" trials. Furthermore, an obstacle course was added on the 11th day of training to reinforce the training. Following training on the 15th day, mice were tested for 3 days to obtain baseline scores for comparison after noise exposure, and then retrained for 3 days.

[0391] After training was complete and mice achieved a high success rate on the trial, they were subjected to noise-induced tinnitus. To ensure that the mice still had hearing, one ear was protected with an earplug while the other ear was exposed to noise. Mice were placed in a soundproof box and exposed to broadband noise at 4-25 kHz and 120 dB for 2-4 hours. After noise exposure, the mice were housed for one month and then tested for the development of tinnitus. During these tests, mice were tested in a shuttle box for three days, undergoing both "Go" and "No-Go" trials. However, mice were not shocked during "No-Go" trials because tinnitus-producing mice were expected to shuttle between compartments in the absence of the audible cue. Mice continued to receive shocks during "Go" trials to ensure that their hearing was preserved and that they could hear the audible cue. When testing results after noise exposure, tinnitus-positive mice had higher average error rates on "No-Go" trials than during training. To ensure significant differences in "No-Go" trial scores, a chi-square test was performed between the post-noise exposure test and the baseline test. If mice did not develop tinnitus, they were retested after one month, or another month if necessary.

[0392] Mice were initially dosed with NMDP via intraperitoneal (ip) injection. "Go" and "No-Go" scores were compared using the chi-square test, a statistical tool used in social sciences. A chi-square test was first used to determine if mice were positive for tinnitus: baseline "No-Go" scores before noise exposure or saline injection were compared with "No-Go" scores after noise exposure. Mice were considered tinnitus-positive if significant differences (p-value < 0.05) were found in at least two tests. Once this conclusion was reached, mice were tested to determine whether different drug doses and combinations could improve their "No-Go" scores. NMDP had no effect on the mice's ability to follow the acoustic cue in the "Go" test. In the "No-Go" test, NMDP did not affect the mice's scores, indicating that concentrations of 10, 30, and 50 mg / kg (ip) did not induce any behavioral changes and should be appropriate test doses.

[0393] Tinnitus was assessed by comparing "No-Go" scores three months after noise exposure to baseline. Mice whose scores were significantly reduced on all three days of testing demonstrated tinnitus in these mice. One or more mice served as control animals. When observing the "Go" scores of tinnitus animals in a specific drug test, NMDP-treated mice showed a significant improvement in their "No-Go" correct response scores, indicating that NMDP is an effective treatment for tinnitus in these mice.

[0394] Figure 1A and 1B Shown are the results of mice injected intraperitoneally with 10, 30, and 50 mg / kg NMDP in the "No-Go" and "Go" tests. Figure 1A The results showed that mice showed only moderate improvement at a dose of 10 mg / kg, which was within the error range of at least one saline control group; at doses of 30 mg / kg and 50 mg / kg, mice showed significant improvement, approaching 100% correct, indicating that NMDP at doses of 30 mg / kg and 50 mg / kg can indeed reduce tinnitus in mice. In addition, Figure 1B The results showed that mice treated with saline, as well as those treated with 10mg / kg and 30mg / kg NMDP, achieved nearly 100% accuracy in the "Go" test, indicating that the mice completed the test as trained and that the experimental data were reliable. The lower accuracy of the 50mg / kg-treated mice in the "Go" test may indicate that they were beginning to experience drug side effects caused by the 50mg / kg dose of NMDP.

[0395] Figure 2A and 2CGraphs depicting the performance of each mouse on the "Go" and "No-Go" tests in each specific test at baseline, saline treatment, 10 mg / kg, 30 mg / kg, and 50 mg / kg NMDP treatment, and treatment with 0.9 mg / mouse of the intranasal formulation of Example 15F1 (or 30 mg / kg based on an average body weight of 30 g at this stage, approximately 5-6 months of age). Figure 2B and 2D is a bar graph showing the percentage of correct answers of mice in “Go” and “No-Go” trials. Figure 2B As shown in the "Go" test, the correct rate of mice treated with normal saline and mice treated with 10mg / kg and 30mg / kg NMDP in the "Go" test was close to 100%, indicating that the mice completed the test according to the training and the experimental data was reliable. The correct rate of mice treated with 50mg / kg and 0.9mg / mouse intranasal injection in the "Go" test was lower, which may indicate that they began to experience some degree of drug side effects. Figure 2C As shown in the "No-Go" test, mice showed only moderate improvement at a dose of 10 mg / kg, and were within the error range of at least one saline control group; at doses of 30 mg / kg, 50 mg / kg and intranasal administration of 0.9 mg / mouse, mice improved significantly, approaching 100% correct, indicating that 30 mg / kg, 50 mg / kg and 0.9 mg / mouse can indeed treat tinnitus in mice.

[0396] also, Figure 2C The correct rate of mice treated intranasally with 50 mg / kg (ip) and 0.9 mg / mouse was closely correlated with the decrease (possibly indicating a side effect of the drug); Figure 2D The results showed that the correct rate of 30 mg / kg, 50 mg / kg and 0.9 mg / mouse was significantly improved (indicating effective treatment of tinnitus), indicating that the effect of 0.9 mg / mouse intranasal NMDP on mice was similar to that of 30 mg / kg and 50 mg / kg intraperitoneal injection. The 0.9 mg intranasal dose of NMDP was only equivalent to about 30 mg / kg of NMDP, which provides strong evidence for the practicality of the intranasal delivery formulation, because its effect was comparable to that of 30 mg / kg and 50 mg / kg intraperitoneal injection, indicating that the dose of NMDP can be significantly reduced when the intranasal formulation is administered.

[0397] Although preferred embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, modifications, and alternatives will now occur to those skilled in the art without departing from the present disclosure. It should be understood that various alternatives to the embodiments of the present disclosure described herein may be employed to implement the present disclosure. It is intended that the following claims define the scope of the present disclosure and encompass methods and structures within the scope of these claims and their equivalents.

Claims

1. A method for treating hearing impairment or a symptom of hearing impairment in a subject in need thereof, comprising intranasally administering to the subject a therapeutically effective amount of an NMDP or L-type calcium channel blocker. The method according to claim 1 , wherein the hearing disorder is tinnitus or Meniere's disease.

3. A method of treating hearing impairment or a symptom of hearing impairment in a subject in need thereof, comprising: a. administering an L-type calcium channel blocker to the subject; b. assessing whether the subject responds to an L-type calcium channel blocker; and c. If the subject is responsive to an L-type calcium channel blocker, administering to the subject a therapeutically effective amount of NMDP or a salt thereof. The method according to claim 3 , wherein the hearing disorder is tinnitus or Meniere's disease.

5. A method for selecting a subject in need of treatment for tinnitus or Meniere's disease or a symptom of tinnitus or Meniere's disease, comprising: a. administering an L-type calcium channel blocker to the subject; b. if the subject responds to the L-type calcium channel blocker, selecting the patient for treatment of tinnitus or Meniere's disease, or a symptom of tinnitus or Meniere's disease; and c. If the subject is responsive to the L-type calcium channel blocker, administering to the subject a therapeutically effective amount of NMDP or a salt thereof.

6. The method of any one of claims 3 or 5, wherein the L-type calcium channel blocker comprises carbamazepine.

7. The method of any one of claims 3 or 5, further comprising administering the therapeutically effective amount of NMDP or a salt thereof intranasally to the patient to treat tinnitus or Meniere's disease, or a symptom of tinnitus or Meniere's disease.

8. The method of claim 7, wherein the therapeutically effective amount of NMDP is administered in a composition comprising: a. the therapeutically effective amount of NMDP; b. carrier; c. Citrate buffer; d. benzalkonium chloride; and e. Ethylenediaminetetraacetic acid.

9. The method of claim 7, wherein the therapeutically effective amount of NMDP is administered in a composition comprising: a. the therapeutically effective amount of NMDP; b. one or more natural or synthetic carriers, or any combination thereof, in an amount of about 10% to about 95% (w / w); and c. a non-aqueous solvent in an amount of about 10% to about 95% (w / w); and d. surfactant.

10. The method of claim 7, wherein the therapeutically effective amount of NMDP is administered in a composition comprising at least one of: a. Polyethylene glycol; b. methoxy polyethylene glycol; and c. Water.

11. The method of any one of claims 1-5, wherein the therapeutically effective amount of NMDP is from about 0.1 mg to about 5 mg per kilogram of the subject.

12. The method of any one of claims 1-11, further comprising administering the pharmaceutical composition in a volume of about 10 microliters to about 300 microliters per dose.

13. The method of any one of claims 1-12, further comprising contacting at least a portion of the therapeutically effective amount of the pharmaceutical composition with a mucous membrane of at least one nasal cavity.

14. The method of any one of claims 1-13, further comprising spraying a first dose of the pharmaceutical composition into a first nasal cavity, spraying a second dose of the pharmaceutical composition into a second nasal cavity, and optionally after a preselected time delay, spraying a third dose of the pharmaceutical composition into the first nasal cavity.

15. The method of claim 14, further comprising administering at least a fourth dose of the pharmaceutical composition to the second nostril, optionally after a preselected time delay.

16. The method of any one of claims 1-15, wherein the method of treatment achieves a bioavailability that is about 80% to 125% of the bioavailability achieved by intravenous administration of the same pharmaceutical composition.

17. The method of any one of claims 1-16, further comprising administering the pharmaceutical composition at any time before or after the onset of symptoms of tinnitus or Meniere's disease.

18. The method of any one of claims 1-17, further comprising administering the pharmaceutical composition at least once a day, twice a day, three times a day, four times a day, five times a day, six times a day, seven times a day, eight times a day, nine times a day, or ten times a day.

19. The method of any one of claims 1-5, wherein the therapeutically effective amount of NMDP is administered intranasally once every 2 hours.

20. The method of any one of claims 1-5, wherein the therapeutically effective amount of NMDP is administered intranasally once every 4 hours.

21. The method of any one of claims 1-5, further comprising administering the pharmaceutical composition weekly.

22. The method of any one of claims 1-12, further comprising administering the pharmaceutical composition via low-dose therapy.

23. The method of any one of claims 1-22, further comprising titrating the pharmaceutical composition for vestibular symptoms.

24. The method of any one of claims 1-23, further comprising administering the pharmaceutical composition for at least 1, 2, 3, 4, 5, 6, 14, 21, 28, 60, 120, or 400 days.

25. The method of any one of claims 1-24, further comprising administering the pharmaceutical composition in a unit dosage form.

26. The method of any one of claims 1-25, further comprising administering the pharmaceutical composition so as not to induce changes in cochlear electrical potentials.

27. The method of any one of claims 1-26, further comprising increasing the subject's dosage of the pharmaceutical composition until symptoms of inner ear dysfunction are observed.

28. The method of claim 27, wherein the inner ear dysfunction symptoms include spontaneous nystagmus, balance disorders, motion intolerance, or hearing loss / decrease observed using Frenzel glasses.

29. The method of any one of claims 1-27, further comprising administering the pharmaceutical composition via a drug delivery device.

30. The method of any one of claims 1-29, wherein the effect of the composition is measured by one of the following assessments: a) measuring changes in ABR and / or DPOAE amplitudes and thresholds, b) changes in auditory speech recognition as measured by a speech-in-noise test, c) changes in auditory speech recognition as measured by a digit-in-noise test, d) changes in low-frequency hearing thresholds, e) changes in the incidence of adverse events following administration of the pharmaceutical composition, f) changes in the severity of tinnitus or Meniere's disease, g) changes in the loudness of tinnitus or Meniere's disease, h) changes in the severity of vertigo, i) changes in aural fullness, j) changes in dizziness, and k) changes in hair cell function as observed by changes in ABR thresholds following administration of the pharmaceutical composition.

31. The method of claim 30, wherein the ABR threshold is measured in the frequency range of 250 Hz to 20 kHz.

32. A method for achieving a therapeutically effective area under the NMDP curve (AUC0- 无穷 ), comprising intranasally administering to the subject an intranasal pharmaceutical composition, wherein the intranasal pharmaceutical composition comprises: (i) about 0.2 to about 250 mg / kg of NMDP or a pharmaceutically acceptable salt thereof; (ii) a buffer; and (iii) surfactants, wherein after administering the intranasal pharmaceutical composition to the subject, the subject's NMDP AUC 0-无穷 Between 270h*ng / mL and 340h*ng / mL.

33. A pharmaceutical composition comprising a therapeutically effective amount of NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a. A therapeutically effective amount of NMDP; b. buffer; c. penetration enhancers; and d. surfactant.

34. The pharmaceutical composition of claim 33, wherein the composition further comprises one or more ingredients selected from the group consisting of vitamin E, vitamin E TPGS, ethanol, benzyl alcohol, and dodecyl maltoside.

35. according to the pharmaceutical composition described in any one in claim 33 or 34, the concentration of NMDP in the pharmaceutical composition wherein administered is 0.5,1,1.5,2,2.5,3,3.5,4,4.5,5,5.5,6,6.5,7,7.5,8,8.5,9,9.5,10,12.5,15,17.5,20,22.5,25,27.5,30,32.5,35,37.5,40,42.5,45,47.5,50,55,60,65,70,75,80,85,90,95,100 mg / ml.

36. The pharmaceutical composition of any one of claims 33-35, wherein the volume of the pharmaceutical composition administered is 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, or 1.0 ml.

37. The pharmaceutical composition according to any one of claims 33 to 35, wherein the pharmaceutical composition is a sustained release formulation.

38. The pharmaceutical composition according to any one of claims 33-35, wherein the pharmaceutical composition is a sustained release formulation.

39. The pharmaceutical composition according to any one of claims 33-35, wherein the pharmaceutical composition is a controlled release formulation.

40. The pharmaceutical composition of any one of claims 33-35, wherein the pharmaceutical composition releases in a continuous, variable or pulsatile manner, or a combination thereof.

41. The pharmaceutical composition of any one of claims 33-35, wherein the pharmaceutical composition is in unit dosage form.

42. The pharmaceutical composition of claim 41, wherein the unit dosage form is a dry powder, a semisolid, a mucoadhesive preparation, an intranasal vesicle unit, or a solution dosage form.

43. The pharmaceutical composition of claim 42, wherein the composition is aqueous.

44. The pharmaceutical composition of claim 43, wherein the composition is in the form of a gel or a film.

45. The pharmaceutical composition of any one of claims 33-44, wherein the composition comprises micronized particles.

46. The pharmaceutical composition of claim 41, wherein the unit dosage form has a unit weight of about 10 mg to about 10 g.

47. The method of claim 42, wherein the unit dose volume of the solution dosage form is less than about 600 μL.

48. The pharmaceutical composition of any one of claims 33-47, comprising NMDP in an amount from about 0.1% to about 20% w / w of the formulation.

49. The pharmaceutical composition of any one of claims 33-48, wherein the pharmaceutical composition prolongs the residence time of the composition in an otic structure.

50. The pharmaceutical composition of claim 49, wherein the formulation extends the residence time of the composition in the otic structure by at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 8 days, or at least 14 days, at least 21 days, or at least 1 month, or at least 6 weeks after a single administration.

51. The pharmaceutical composition of claim 33, wherein the formulation increases the bioavailability of the composition in otic structures.

52. The pharmaceutical composition of claim 51, wherein the formulation increases steady-state levels of the composition in otic structures.

53. The pharmaceutical composition of claim 52, wherein the formulation increases the C to a therapeutic concentration capable of alleviating symptoms of hearing impairment in a subject in need thereof. max time.

54. The pharmaceutical composition according to claim 53, wherein the formulation prolongs the maintenance of the concentration of the composition at the minimum therapeutic concentration (i.e., C min ) or more time.

55. The pharmaceutical composition of claim 54, wherein the concentration of the composition in the otic structure is maintained at or above the concentration of Cmin for at least 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days, 3 weeks, or 1 month.

56. The pharmaceutical composition of any one of claims 33-55, further comprising a second pharmaceutically active agent.

57. A pharmaceutical composition comprising a therapeutically effective amount of NMDP, the composition being formulated for intranasal administration, the composition comprising: a. a therapeutically effective amount of NMDP; b. a surfactant; c. a solvent; and d. a water-soluble cellulose polymer.

58. The pharmaceutical composition of claim 57, further comprising water.

59. The pharmaceutical composition of any one of claims 57-58, further comprising ethanol.

60. The pharmaceutical composition of any one of claims 57-59, wherein the ethanol is at most 5% (w / w).

61. The pharmaceutical composition of any one of claims 57-60, wherein the surfactant is a polysorbate or a combination of polysorbates.

62. The pharmaceutical composition of any one of claims 57-61, wherein the surfactant is polysorbate 20 (TWEEN-20).

63. The pharmaceutical composition of any one of claims 57-61, wherein the surfactant is polysorbate 80 (TWEEN-80).

64. The pharmaceutical composition of any one of claims 57-61, wherein the surfactant is at least 0.00001% and at most 2% (w / w).

65. The pharmaceutical composition of any one of claims 57-64, wherein the surfactant is at least 0.05% (w / w) and at most 2% (w / w).

66. The pharmaceutical composition of any one of claims 57-65, wherein the water-soluble cellulose polymer is HPMC, HEC, CMC, sodium carboxymethylcellulose, or a combination thereof.

67. The pharmaceutical composition of any one of claims 57-66, wherein the water-soluble cellulose polymer is at least 0.00001% and at most 2% (w / w).

68. The pharmaceutical composition of any one of claims 57-67, wherein the water-soluble cellulose polymer is at least 0.05% (w / w) and at most 2% (w / w).

69. The pharmaceutical composition of any one of claims 57-68, wherein the solvent is PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof.

70. The pharmaceutical composition of any one of claims 57-69, wherein the solvent is at least 0.00001% and at most 94% (w / w).

71. The pharmaceutical composition of any one of claims 57-69, wherein the solvent is at least 66% (w / w) and at most 94% (w / w).

72. The pharmaceutical composition of any one of claims 57-71, wherein the water is at least 0.00001% and at most 50% (w / w).

73. The pharmaceutical composition of any one of claims 57-72, wherein the water is at least 0.00001% and at most 30% (w / w).

74. The pharmaceutical composition of any one of claims 57-73, wherein the water is at least 5% (w / w) and at most 50% (w / w).

75. The pharmaceutical composition of any one of claims 57-74, wherein the water is at least 5% (w / w) and at most 30% (w / w).

76. A pharmaceutical composition comprising a therapeutically effective amount of NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a. a therapeutically effective amount of NMDP; b. a surfactant; c. a water-soluble cellulosic polymer; d. a solvent; and e. water.

77. The pharmaceutical composition of claim 76, wherein the pharmaceutical composition further comprises three solvents.

78. The pharmaceutical composition of any one of claims 76-77, wherein the three solvents comprise at least three PEGs, alkoxy derivatives of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof.

79. The pharmaceutical composition of any one of claims 76-78, wherein the surfactant is a polysorbate or a combination of polysorbates.

80. The pharmaceutical composition of any one of claims 76-79, wherein the surfactant is polysorbate 20 (Tween-20).

81. The pharmaceutical composition of any one of claims 76-79, wherein the surfactant is polysorbate 80 (Tween-80).

82. The pharmaceutical composition of claim 81, wherein the polysorbate 80 (Tween-80) is at least 0.00001% and at most 2% (w / w).

83. The pharmaceutical composition of claims 81-82, wherein the polysorbate 80 (Tween-80) is at least 0.05% (w / w) and at most 2% (w / w).

84. The pharmaceutical composition of any one of claims 73-83, wherein the water-soluble cellulose polymer is HPMC, HEC, CMC, sodium carboxymethylcellulose, or a combination thereof.

85. The pharmaceutical composition of any one of claims 73-84, wherein the water-soluble cellulose polymer comprises only HPMC.

86. The pharmaceutical composition of claims 76-85, wherein the HPMC is at least 0.00001% and at most 2% (w / w).

87. The pharmaceutical composition of claims 76-86, wherein the HPMC is at least 0.05% (w / w) and at most 2% (w / w).

88. The pharmaceutical composition of any one of claims 73-87, wherein the solvent is PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof.

89. The pharmaceutical composition of any one of claims 73-88, wherein the PEG, alkoxy derivatives of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof is at least 0.00001% and at most 94% (w / w).

90. The pharmaceutical composition of any one of claims 73-89, wherein the PEG, alkoxy derivatives of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof is at least 66% (w / w) and at most 94% (w / w).

91. The pharmaceutical composition of any one of claims 73-90, wherein the water is at least 0.00001% and at most 50% (w / w).

92. The pharmaceutical composition of any one of claims 73-91, wherein the water is at least 5% (w / w) and at most 50% (w / w).

93. The pharmaceutical composition of any one of claims 73-92, wherein the water is at least 0.00001% and at most 30% (w / w).

94. The pharmaceutical composition of any one of claims 73-93, wherein the water is at least 5% (w / w) and at most 30% (w / w).

95. A method for preparing a liquid NMDP composition, the method comprising the steps of: 1) Dissolve Tween 80 (<2% w / w) in mPEG350 and PEG400, respectively; 2) mixing NMDP with the solution of step 1) to saturated solubility or desired effective concentration; 3) preparing an aqueous solution of HPMC or a solution of HPMC in a buffer (<2% w / w) with a pH in the range of about 6.4 to about 7.4; and 3) Adding the HPMC aqueous solution or HPMC buffer of step 3) to the corresponding solution of step 2) under stirring, and allowing the mixture to stand to observe the stability of the solution or suspension.

96. The method according to claim 95, wherein the solution in step 2) is NMDP in PEG / Tween 80 solution or NMDP in mPEG / Tween 80 solution.

97. The pharmaceutical composition of any one of claims 57-96, wherein the concentration of NMDP is at least 68 mg / mL.

98. The pharmaceutical composition of any one of claims 57-97, wherein the concentration of NMDP is at least 0.1 mg / mL to 1 mg / mL.

99. The pharmaceutical composition of any one of claims 57-98, wherein the concentration of NMDP is 68 mg / mL.

100. The pharmaceutical composition of any one of claims 57-99, wherein the concentration of NMDP is 50 mg / mL.

101. The pharmaceutical composition of any one of claims 57-100, wherein the surfactant, water-soluble cellulosic polymer, solvent, water, and ethanol comprise a liquid carrier.

102. The pharmaceutical composition of any one of claims 57-101, wherein the liquid carrier is less than 1000 uL.

103. The pharmaceutical composition of any one of claims 57-102, wherein the liquid vehicle is less than 300 uL.

104. The pharmaceutical composition of any one of claims 57-103, wherein the liquid vehicle is less than 150 uL.

105. The pharmaceutical composition of any one of claims 57-104, wherein the NMDP is dissolved, suspended, or both dissolved and suspended in the liquid carrier.

106. A pharmaceutical composition comprising a therapeutically effective amount of an NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a. a therapeutically effective amount of NMDP; and b. a liquid carrier, wherein the liquid carrier further comprises i. surfactants; ii. water-soluble cellulose polymers; iii. solvent; and iv. Water.

107. The pharmaceutical composition of claim 76 or 96, wherein: a. The liquid carrier further comprises: i. the surfactant; ii. the water-soluble cellulose polymer; iii. the solvent; and iv. said water.

108. A pharmaceutical composition comprising a therapeutically effective amount of an NMDP, wherein the composition is formulated for intranasal administration, the composition comprising: a. A therapeutically effective amount of NMDP; b. solvent; and c. Water.

109. The pharmaceutical composition of claim 108, wherein the solvent is PEG, an alkoxy derivative of polyethylene glycol, mPEG, mPEG350, PEG400, or a combination thereof.

110. The pharmaceutical composition of any one of claims 108-109, wherein the solvent is at least 0.00001% and at most 94% (w / w).

111. The pharmaceutical composition of any one of claims 108-110, wherein the solvent is at least 66% (w / w) and at most 94% (w / w).

112. The pharmaceutical composition of any one of claims 108-111, wherein the water is at least 0.00001% and at most 20% (w / w).

113. The pharmaceutical composition of claim 108, wherein the composition further comprises a first solvent and a second solvent.

114. The pharmaceutical composition of claim 113, wherein the first solvent comprises mPEG and the second solvent comprises PEG.

115. The pharmaceutical composition of claim 114, wherein the mPEG comprises mPEG350.

116. The pharmaceutical composition of claim 114, wherein the PEG comprises PEG400.

117. The pharmaceutical composition of claim 108, wherein the therapeutically effective amount of NMDP comprises at least 10 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / mL, at least 30 mg / mL, at least 35 mg / mL, at least 40 mg / mL, at least 45 mg / mL, at least 50 mg / mL, at least 55 mg / mL, at least 60 mg / mL, at least 65 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL, at least 95 mg / mL, at least 100 mg / mL, at least 110 mg / mL, or greater than 110 mg / mL.

118. The pharmaceutical composition of claim 114, wherein the mPEG comprises about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, or about 80% to about 90% (w / w) of the composition, the PEG comprises about 10% to about 50%, about 20% to about 50%, about 30% to about 50%, or about 40% to about 50% (w / w) of the composition, and the water comprises about 5% to about 20%, about 10% to about 20%, or about 15% to about 20% (w / w) of the composition.

119. The pharmaceutical composition of claim 108, wherein the composition further comprises ethanol.

120. The pharmaceutical composition of claim 119, wherein the ethanol comprises about 1% to about 10%, about 2% to about 10%, about 3% to about 10%, about 4% to about 10%, about 5% to about 10%, about 6% to about 10%, about 7% to about 10%, about 8% to about 10%, or about 9% to about 10% of the composition.

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