Novel formulations of adrenaline and uses thereof

Abstract

The present disclosure relates to novel formulations of epinephrine comprising polyoxyethylene alkyl ethers as penetration enhancers, methods of administration of the formulations, and uses thereof.

Description

Invention Field

[0001] The present invention relates to a novel adrenaline formulation containing polyoxyethylene alkyl ether as a penetration enhancer, a method of administration of the formulation, and its use.

[0002] Cross-reference to related applications

[0003] This application claims priority and benefit from international patent application PCT / CN2022 / 126776, filed on October 21, 2022, the entire contents of which are incorporated herein by reference.

[0004] background

[0005] Adrenaline is a hormone and neurotransmitter secreted by the adrenal glands. It can also be chemically synthesized. Adrenaline is used as a drug to treat a variety of conditions.

[0006] Specifically, adrenaline is the first-line drug for treating type I anaphylactic reactions, including anaphylactic shock. It is estimated that up to 2% of the world’s population will experience an anaphylactic reaction at some point in their lives, and this is on the rise (Simonset et al., The World Allergy Organization Journal, 4(2):13–37 (2006)). Anaphylactic shock is a severe, potentially fatal anaphylactic reaction and medical emergency that develops rapidly and requires immediate medical attention, regardless of whether emergency medications are used on-site (Sampson et al., J. Allergy and Clinical Immunology, 117(2):391–7; Tintinalli, Judith E., Emergency Medicine: A Comprehensive Study Guide (2010) New York: McGraw-Hill Companies.pp.177–182. ISBN 978-0-07-148480-0). Therefore, an auto-injector for adrenaline was developed to allow for faster self-administration of adrenaline via intramuscular injection in emergency situations (Mylan Specialty LP). -Epinephrine injection, EPIPEN - Epinephrine Injection (FDA product label (archived February 1, 2014 (PDF), retrieved January 22, 2014)). However, epinephrine auto-injectors have a higher probability of causing dangerous situations such as misuse as subcutaneous or intravenous injection, or use of incorrect dosage (Bilò, M. Beatrice., Anaphylaxis caused by Hymenoptera stings: from epidemiology to treatment, Allergy 66, pages 35-37 (2011)). The structural complexity of epinephrine injectors also makes them prone to mechanical failures, reducing the reliability of this life-saving drug as expected. In addition, patients (whether or not they have needle phobia) are reluctant to use auto-injectors in public places and often delay using them until their condition becomes severe, thus delaying treatment and ultimately leading to the inability to effectively reverse the rapid progression of type I hypersensitivity reactions.

[0007] Given the challenges faced by auto-injectors for epinephrine, there is an urgent need for more reliable and patient-compliant epinephrine delivery methods. Nasal administration is one of the promising routes of administration, and several nasal spray formulations of epinephrine are currently under development. However, existing nasal spray formulations of epinephrine are plagued by low absorption, nasal mucosal damage, and poor chemical stability. For example, compared to intramuscular injection, even with the use of penetration enhancers, existing nasal spray formulations of epinephrine still require a dose greater than three times (0.3 mg vs 1 mg) (Australian Patent No. AU2019217643B2). It is well known that epinephrine has a very narrow therapeutic window and poor absorption, which is often associated with large variability in absorption rates. Therefore, when using higher doses to address poor absorption, it may lead to overdose in patients with high absorption rates, resulting in serious adverse reactions such as cerebral hemorrhage, hemiplegia, subarachnoid hemorrhage, and respiratory distress. Similar situations have been identified as a major safety risk requiring further improvement for another drug (Hayley B. Schultz, et al., Oral Formulation Strategies to Improve the Bioavailability and Mitigate the Food Effect of Abiraterone Acetate, International Journal of Pharmaceutics, Volume 577, 119069, 2020). Furthermore, the European Medicines Agency (EMA) has explicitly stated that early partial AUC values, particularly the AUC within the first 10 minutes after administration, are crucial to the efficacy of epinephrine nasal sprays; Tmax It also has a correlation; at the same adrenaline dose, T max It should be equivalent to intramuscular (IM) or subcutaneous (SC) injection; C max Total AUC is considered to be the most relevant parameter to drug safety and efficacy. Neffy is an adrenaline nasal spray that submitted a New Drug Application (NDA) to the EMA, but was not approved due to its lower early-stage AUC compared to intramuscular injection (IM) (Assessment Report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0008] Furthermore, reports indicate that the simultaneous use of penetration enhancers and epinephrine in nasal spray formulations can lead to severe damage to the nasal mucosa (Bleske et al., Effect of Vehicle on the Nasal Absorption of Epinephrine during Cardiopulmonary Resuscitation, Pharmacotherapy, 16(6), 1039–1045 (1996)). In addition, epinephrine is highly susceptible to oxidation, making the maintenance of the stability of epinephrine formulations extremely challenging (GBWest, Oxidation of Adrenaline in Alkaline Solution, British Journal of Pharmacology and Chemotherapy, Volume 2, Issue 2, p. 121-130, 1947). Therefore, there is a need to develop an epinephrine nasal spray formulation with higher bioavailability, less nasal mucosal damage, less individual variability, faster absorption rate, and better chemical stability. Summary of the Invention

[0009] This invention discloses a novel adrenaline formulation comprising polyoxyethylene alkyl ether as a penetration enhancer, a method of administration of the adrenaline formulation, and its uses. The formulation disclosed in this invention can have higher bioavailability, faster absorption rate, better chemical stability, and less damage to the nasal mucosa.

[0010] On the one hand, this invention discloses a pharmaceutical composition containing adrenaline and a penetration enhancer having the following formula: CH3(CH2) n-1 [OCH2CH2] mOH, where n is an integer selected from 10 to 16, and m is an integer selected from 4 to 8.

[0011] In some implementations, n equals 12, and m is selected from 4, 7, and 8.

[0012] In some implementations, n equals 10 and m equals 6.

[0013] In some embodiments, the concentration of the penetration enhancer in the pharmaceutical composition described herein ranges from 0.1% to 2.50% (v / v).

[0014] In some embodiments, the concentration of the penetration enhancer in the pharmaceutical composition described herein is 0.25% (v / v).

[0015] In the pharmaceutical composition of any one of claims 1-5, the pH value of the pharmaceutical composition is less than 7.

[0016] In the pharmaceutical composition of any one of claims 1-6, the pH value of the pharmaceutical composition ranges from 4 to 6.

[0017] In some embodiments, the pharmaceutical composition is used for nasal administration or for use in a nasal administration device.

[0018] In some embodiments, the pharmaceutical composition further includes at least one pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is a pH adjuster, antioxidant, preservative, or osmotic pressure regulator.

[0019] In some embodiments, the antioxidant is selected from sodium bisulfite, sodium metabisulfite (SMB), propyl gallate (PG), sodium sulfite, ascorbic acid (VC), methionine, alpha-lipoic acid, cysteine ​​(CYS), D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS), butylated hydroxytoluene (BHT), and butylated hydroxyanisole (BHA).

[0020] In some embodiments, the concentration of adrenaline in the pharmaceutical composition is in the range of 0.3% to 5% (w / v).

[0021] In some embodiments, the pharmaceutical composition is in the form of a liquid or a spray.

[0022] In some embodiments, the recovery rate of the pharmaceutical composition on day 30 at pH 4.0 and 60°C is not less than 90% by weight.

[0023] In some implementations, the effective penetration coefficient (Pe) of the pharmaceutical composition is tested using PAMPA and is greater than 3 × 10⁻⁶. - 6 cm / s.

[0024] In some embodiments, the pharmaceutical compositions disclosed in this invention do not cause irreversible damage to the nasal mucosa.

[0025] On the other hand, the present invention discloses a method comprising administering an effective amount of a pharmaceutical composition to a test subject.

[0026] In some implementations, this method is used to treat allergic reactions in test subjects, particularly type I allergic reactions.

[0027] In some implementations, the allergic reaction is selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, anaphylactic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

[0028] In some embodiments, the pharmaceutical composition is administered to the test subject via nasal administration.

[0029] Brief description of the attached figures

[0030] Figure 1 This shows the change in plasma adrenaline concentration over time in the group with an adrenaline concentration of 10 mg / mL (1 mg / kg dose).

[0031] Figure 2 This shows the change in plasma adrenaline concentration over time in the 3 mg / mL adrenaline concentration group (3 mg / mL adrenaline concentration is much lower than the clinically used concentration, with a dose of 0.3 mg / kg).

[0032] Figure 3 This study shows the changes in plasma adrenaline concentration over time after administration of compositions with different adrenaline concentrations and different types and concentrations of osmotic enhancers.

[0033] Figure 4 This study shows the changes in plasma adrenaline concentration over time after administration of compositions with different adrenaline concentrations and different types of osmotic enhancers.

[0034] Figure 5 The C values ​​for each subgroup of each treatment group are shown. max (μU / mL).

[0035] Figure 6 The AUC (min*μU / mL) of each subgroup in each treatment group is displayed.

[0036] Detailed description

[0037] definition

[0038] In this disclosure, unless otherwise specified, the technical terms used herein have the meanings commonly understood by those skilled in the art. Accordingly, the terms defined herein are described more fully with reference to the entire specification.

[0039] As used in this article, unless otherwise specified, the absence of a plural quantifier before an element also includes the meaning of plural.

[0040] As used herein, “and / or” means and includes any and all possible combinations of one or more of the listed items, as well as no combination when understood as an alternative form (“or”). Furthermore, the invention also contemplates that in some disclosed embodiments, any feature or combination of features described herein may be excluded or omitted.

[0041] Unless otherwise stated in the text, the terms “contains,” “comprising,” or similar terms are intended to indicate non-exclusionary inclusion, and therefore the list of elements or features described includes not only those explicitly identified or listed, but may also include other elements or features that are not listed or explicitly identified.

[0042] It should be understood that the disclosure of this invention is not limited to the specific methods, schemes and reagents described, and the above factors may vary depending on the circumstances in which those skilled in the art use them.

[0043] As used herein, “systemic administration” is defined as a route of administration of a therapeutic product that produces widespread exposure of the active agent in a living organism. As used herein, “local administration” refers to a route of administration that delivers the active agent directly to the target site in a living organism. Local administration does not exclude systemic pharmacological effects.

[0044] As used herein, the term "pharmaceutically acceptable" means that a material, such as a carrier or diluent, does not diminish the biological activity or properties of a therapeutic compound and is relatively non-toxic; that is, the material can be applied to a test subject without causing adverse biological effects or having a harmful effect on any component of a composition containing the material. Pharmaceutically acceptable components include compounds, materials, compositions, and / or dosage forms that are suitable for use in human and animal tissues, within reasonable medical judgment, without excessive toxicity, irritation, allergic reactions, or other problems or complications, and in proportion to a reasonable benefit / risk ratio.

[0045] As used herein, “effective amount” means an amount of pharmaceutical composition sufficient to significantly and positively alter the symptoms and / or condition to be treated (e.g., provide a positive clinical response). The effective amount of a pharmaceutical composition will vary depending on factors such as the specific condition being treated, the severity of the condition, the duration of treatment, the nature of concurrent treatments, the specific composition used, the specific pharmaceutically acceptable excipients and / or carriers used, and the knowledge and experience of the physician.

[0046] As used in this article, "disease" or "disorder" refers to a condition that requires and / or is expected to be treated.

[0047] As used herein, the term "treatment" or "therapy" refers to improving a disease or condition, such as slowing, halting, or reducing the development of a disease or condition or reducing at least one of its clinical symptoms. For example, in some embodiments, improving a disease or condition may include obtaining beneficial or desired clinical outcomes, including but not limited to one or more of the following: relieving one or more symptoms, reducing the severity of the disease, halting or delaying the spread of the disease, halting or delaying the recurrence of the disease, delaying or slowing the progression of the disease, improving the disease state, inhibiting the disease or its progression, inhibiting or slowing the disease or its progression, halting its development, and alleviating (whether partially or completely).

[0048] As used herein, the term "subject" refers to an animal. For example, in some embodiments, the animal is a mammal. In some embodiments, the animal includes humans, rodents, primates, felines, canines, equines, bovines, suidae, sheep, goats, laboratory mammals, farm mammals, livestock mammals, or pet mammals. The animal can be male or female and can be of any suitable age, including infants, young children, adolescents, adults, and the elderly. In some embodiments, "individual" or "subject" refers to an animal that requires treatment for a disease or condition. In some embodiments, the animal to be treated may be referred to as a "patient," meaning an animal that has been identified as having a treatment-related disease or is at sufficient risk of developing the disease. In some embodiments, this animal is a human, such as a human patient.

[0049] As used herein, “permeability” refers to the ability of a pharmaceutical composition to pass through a biological membrane. In some embodiments, the biological membrane is the nasal mucosa. Permeability can be measured using various permeability models, which can be performed in situ, in vivo, or in vitro. Some exemplary permeability models will be discussed in later sections.

[0050] As used herein, the term "penetration enhancer" refers to an excipient contained in a formulation for improving the permeability of the active pharmaceutical ingredient. Sometimes "penetration enhancer" is also referred to as "absorption enhancer" or "permeability enhancer." In some embodiments, a penetration enhancer promotes nasal mucosal permeability. In some embodiments, a penetration enhancer promotes the passage of bypass pathways. In some embodiments, a penetration enhancer promotes the passage of transcellular channels.

[0051] As used herein, the terms "nasal administration" or "intranasal administration" refer to the application of a pharmaceutical composition into the nasal cavity of a test subject for local or systemic administration. As used herein, "nasal" and "intranasal" are used interchangeably.

[0052] As used herein, the terms “allergic reaction,” “allergic reaction,” “allergy,” and “hypersensitivity” are used interchangeably.

[0053] As used herein, in the context of pharmaceutical compositions, the term "active ingredient" refers to any component that provides pharmacological activity or other direct action for the diagnosis, cure, relief, treatment, or prevention of disease, or affects the structure or any function of a subject's body. For the purposes of this disclosure, the active ingredient of the adrenaline compositions described herein is adrenaline.

[0054] As used in this article, the term "therapeutic window" refers to the range of blood concentrations of a drug within which the desired effect occurs, below which there is little effect, and above which excessive toxicity occurs.

[0055] Overview

[0056] Epinephrine is a first-line treatment for type I anaphylaxis, also known as immediate anaphylaxis, which involves the release of immunoglobulin E (IgE)-mediated antibodies against soluble antigens, leading to mast cell degranulation and the release of histamine and other inflammatory mediators. Type I anaphylaxis includes atopic diseases, which are excessive IgE-mediated immune responses (e.g., rhinitis, conjunctivitis, and dermatitis); and allergic diseases, which are immune responses to exogenous allergens (e.g., anaphylactic shock, urticaria, angioedema, and food and drug allergies). By binding to multiple receptors on cells, epinephrine helps increase blood flow, relax lung muscles, and inhibit the release of chemicals that trigger anaphylactic reactions. At specific doses and routes of administration, alpha-adrenergic vasoconstriction reverses peripheral vasodilation, thereby relieving hypotension and reducing erythema, urticaria, and angioedema. The β-adrenergic properties of adrenaline lead to bronchodilation, increased myocardial output and contractility, and inhibition of further mediator release from mast cells and basophils.

[0057] When a patient experiences a type I anaphylactic reaction, epinephrine needs to be administered rapidly. However, due to patient fear of needles, lack of training, and misunderstandings about the correct timing of administration, intramuscular administration of epinephrine can be significantly delayed. This delay can lead to adverse outcomes and even death. Furthermore, intramuscular administration of epinephrine may result in it being mistakenly administered intravenously or subcutaneously, causing serious side effects and significantly delaying the onset of action. Nasal administration overcomes these problems, providing patients with a more convenient and timely way to administer epinephrine.

[0058] Nasal drug delivery is a non-invasive route of administration in which a drug composition is inhaled through the nasal cavity and absorbed through the nasal mucosa. The drug must first penetrate the nasal mucosal layer and then the epithelial layer to be absorbed and achieve a systemic effect. Drugs administered nasally can permeate passively via cellular bypass pathways, or via both passive and active transcellular pathways. This process is largely influenced by the lipophilicity of the compound. Besides passive transport pathways, carrier-mediated transport, transmembrane transport, and transport through intercellular tight junctions are other possible pathways for drug penetration through the nasal mucosa (Arora et al., Permeability Issues in Nasal Drug Delivery, Drug DiscoveryToday Vol. 7-18, 2002). For polar and hydrophilic drugs, such as adrenaline, the primary pathway is the cellular bypass pathway, which involves the intercellular space and tight junctions.

[0059] Two major obstacles to drug absorption in nasal administration are the low membrane permeability of polar drugs and the rapid clearance rate by the nasal mucosa and cilia. Therefore, finding more effective penetration enhancers for intranasal formulations is crucial to achieving systemic absorption and pharmacokinetics comparable to intramuscular injection. This invention provides such effective penetration enhancers and their uses.

[0060] Penetration enhancer

[0061] Polyoxyethylene alkyl ethers are nonionic surfactants composed of a linear alkyl chain (containing n-1 methylene groups) and a hydrophilic moiety (containing m oxyethylene units). They have the common chemical formula CH3(CH2). n-1 (OCH2CH2) m OH (chemical formula I). ​​They are also known as CnEm, where n represents the number of carbons in the alkyl chain and m represents the number of ethylene oxide units in the hydrophilic part.

[0062]

[0063] Although polyoxyethylene alkyl ethers have been nominated as penetration enhancers in previous publications, they constitute a large family containing an infinite number of compounds, few of which are effective and safe for use as penetration enhancers in any commercially available nasal formulation. For example, nasal administration of polyoxyethylene-9-lauryl ether (C12E9) at a concentration of 1% has been reported to cause severe multiple erosions of the nasal epithelium in dogs, suggesting that polyoxyethylene alkyl ethers may not be suitable for nasal administration of epinephrine (Bleske et al., Effect of Vehicle on the Nasal Absorption of Epinephrine During Cardiopulmonary Resuscitation, Pharmacotherapy 1996; 16(6):1039-1045). However, the current study has unexpectedly found that certain types of polyoxyethylene alkyl ethers exhibit significantly higher bioavailability, shorter onset of action, better safety, and chemical stability in nasal administration of epinephrine compared to other formulations using C12E9 and other types of penetration enhancers.

[0064] On one hand, the present invention discloses a pharmaceutical composition comprising adrenaline and a penetration enhancer having the following formula: CH3(CH2) n-1 [OCH2CH2] m OH, where n is an integer selected from 10, 11, 12, 13, 14, 15 and 16, and m is an integer selected from 4, 5, 6, 7 and 8.

[0065] In some implementations, n is 12, and m is selected from 4, 7, and 8.

[0066] In some implementations, n is 10 and m is 6.

[0067] For the purposes of this invention, adrenaline includes the free form of adrenaline and pharmaceutically acceptable salts of adrenaline, including acid addition salts and base addition salts.

[0068] "Pharmaceutically acceptable acid addition salts" refer to salts that retain the bioavailability and properties of the free base without acquiring biological or other adverse properties, formed by the reaction of a free base with an inorganic or organic acid. Some exemplary acids include hydrochloric acid, tartaric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid.

[0069] "Pharmaceutically acceptable base addition salts" refer to salts that retain the bioavailability and properties of the free acid without acquiring any biological or other adverse properties. These salts are prepared by adding an inorganic or organic base to the free acid. Salts prepared from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts.

[0070] The invention disclosure provided herein is intended to include all pharmaceutically acceptable compounds described herein that are isotopically labeled by replacing one or more atoms with atoms having different atomic masses or mass numbers. Examples of isotopes that can be introduced into said compounds include isotopes of hydrogen, carbon, and oxygen, such as 2H, 3H, 11C, 13C, 14C, 15O, 17O, and 18O.

[0071] In some embodiments, the concentration of the penetration enhancer ranges from 0.1% to 2.50% (v / v). In some embodiments, the concentration of the penetration enhancer is 0.25% (v / v). In some embodiments, the concentration of the penetration enhancer is about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, or 2.5% (v / v).

[0072] As used herein, “w / v” means weight volume, also known as mass volume. Weight volume concentration is calculated by dividing the mass of the solute by the volume of the solution; throughout this application, grams of solute per 100 mL of solution (g / 100 mL) are used. In this application, “v / v” means volumetric volume. Volumetric volume concentration is calculated by dividing the volume of the solute by the volume of the solution. For the purposes of this invention, when a concentration is expressed as “w / v or v / v”, this means that for solid solutes, the concentration is w / v, and for liquid solutes, the concentration is v / v. For compounds that are solid at room temperature but have a low melting point, such as C12E8, their v / v concentration is obtained by first melting the compound and then preparing a solution.

[0073] It is understood that those skilled in the art can convert w / v to v / v based on the density of the solute compound, and vice versa. The density of C10E6 is approximately 0.987 g / mL, the density of C12E4 is approximately 0.946 g / mL, the density of C12E7 is approximately 1.0 g / mL, the density of C12E8 is approximately 0.984 g / mL (measured at 35°C), and the density of C12E9 is approximately 1.007 g / mL.

[0074] In some embodiments, the pharmaceutical composition is used for nasal administration or for use in a nasal administration device. In some embodiments, nasal administration is performed by applying the pharmaceutical composition directly to the nasal mucosa. In some embodiments, nasal administration is performed by a subject inhaling the pharmaceutical composition into the nasal cavity.

[0075] The nasal cavity is divided into the vestibule, vestibular cavity, inferior turbinate, middle turbinate, and superior turbinate. After nasal administration, drug deposition mainly occurs in the respiratory area surrounding the inferior turbinate (Grassin-Delyle et al., Pharmacology & Therapeutics 134:366-379 (2012)). The nasal mucosa consists of the epithelial cell layer covering the nasal cavity. Drugs can be absorbed into the systemic circulation through the nasal mucosa.

[0076] Some examples of nasal delivery devices include vapor inhalers, droppers, pipettes, squeeze bottles, spray pumps, nebulizers, powder sprayers, and blowpipes (Djupesland, Drug Deliv. And Transl. Res. 3:42-62, 2013). It will be understood that those skilled in the art can select a suitable nasal delivery device based on the dosage form, chemical properties, physical properties, and other relevant considerations of a given pharmaceutical composition.

[0077] In some embodiments, the concentration of adrenaline in the pharmaceutical composition described herein ranges from 0.3% to 5% (w / v). In some embodiments, the concentration of adrenaline in the pharmaceutical composition described herein is about 0.3%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5% (w / v).

[0078] In some embodiments, the pharmaceutical composition described in this invention is in the form of a liquid. In some embodiments, the pharmaceutical composition described in this invention is in the form of a spray. In some embodiments, the pharmaceutical composition forms water droplets.

[0079] Permeability

[0080] As used herein, “permeability” refers to the ability of a pharmaceutical composition to pass through a biological membrane. For the purposes of this disclosure, the effective permeability coefficient (Pe) is used as an indicator of in vitro permeability testing in parallel artificial membrane permeability assays (PAMPA). PAMPA permeability testing is based on the passive diffusion of the target compound through the artificial membrane. The PAMPA artificial membrane has a lipid-oil-lipid sandwich structure constructed in the pores of a porous filter. The intermediate oil layer maintains a robust and stable PAMPA membrane and is ultrathin to minimize the retention of the compound and interference with the permeation of the compound. The test compound diluted in buffer is placed at the supply end, and the compound enters the artificial membrane from the supply end and enters the receiver end by passive diffusion (Kansy et al., Drug Discov Today Technol, 1(4):349-55 (2004); Avdeef, Expert Opin Drug Metab Toxicol., 1(2):325-42 (2005); Kerns et al., J Pharm Sci., 93(6):1440-53 (2004)). The Pe value is used to determine the permeation rate. In PAMPA, the larger the absolute value of Pe, the better the in vitro permeability. The detailed scheme of PAMPA is described in Example 1.

[0081] The penetration enhancer disclosed in this invention can effectively improve the permeability of adrenaline compositions. In some embodiments, the pharmaceutical composition described in this invention has an absolute Pe value greater than 3 × 10⁻⁶ in the PAMPA test. -6 cm / s.

[0082] Furthermore, when the pharmaceutical composition is administered to a test subject, the maximum plasma concentration (C0) of the active ingredient of the pharmaceutical composition is... max ) and reach C max Required time (T) max ( ) are two effective indicators of permeability in the body. C max The larger the value, the better the permeability within the body. max The smaller the value, the better the permeability within the body.

[0083] Pharmacokinetic characteristics

[0084] Compared to previously disclosed penetration enhancers, namely C12E9, n-dodecyl-β-D-maltose glycoside (DDM), and diethylene glycol monoethyl ether (DEGEE), the penetration enhancers disclosed in this invention exhibit more desirable pharmacokinetic characteristics. In some embodiments, the pharmaceutical compositions disclosed in this invention exhibit better bioavailability. In some embodiments, the pharmaceutical compositions disclosed in this invention exhibit a faster absorption rate, i.e., a shorter T0. maxIn some embodiments, the pharmaceutical compositions disclosed in this invention exhibit a larger area under the curve (AUC) during the early partial drug-time period.

[0085] This invention disclosure also provides an unexpected finding: the pharmaceutical compositions described herein exhibit higher bioavailability compared to previously disclosed penetration enhancers (such as C12E9, DDM, and DEGEE), particularly at higher epinephrine doses closer to clinical use. Some pharmaceutical compositions described herein even exhibit higher bioavailability. For example, nasal administration of an epinephrine composition containing C12E7 as a penetration enhancer results in 2.7 times higher exposure compared to intramuscular epinephrine administration.

[0086] Compared to previously disclosed compositions (such as C12E9, DDM, and DEGEE), the compositions disclosed in this invention exhibit shorter To. max And a larger area under the curve (AUC) in the early stages of drug delivery. A shorter T0 max A larger area under the curve (AUC) in the early part of the drug is a more advantageous characteristic because adrenaline is a life-saving drug used in emergency situations, and faster adrenaline onset is highly desirable (Assessment report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0087] Security

[0088] Compared to previously disclosed penetration enhancers (such as C12E9, DDM, and DEGEE), the penetration enhancers disclosed in this invention exhibit superior safety. In some embodiments, the pharmaceutical compositions disclosed in this invention demonstrate better tolerability. In some embodiments, the pharmaceutical compositions disclosed in this invention do not cause irreversible damage to the nasal mucosa.

[0089] Epinephrine has a narrow therapeutic window. Doses exceeding this window can lead to fatal side effects such as cerebral hemorrhage and subarachnoid hemorrhage. Therefore, an ideal penetration enhancer should improve tolerability to epinephrine. In the disclosed pharmacokinetic studies, all subjects in the C12E9 group who received intranasal administration of an epinephrine composition containing C12E9 (1 mg / kg dose) died, with only 53.2% bioavailability compared to intramuscular injection. However, after intranasal administration of an epinephrine composition containing C12E7 (1 mg / kg dose), only one subject experienced shock (recovered after cardiopulmonary resuscitation), with a bioavailability exceeding 270% compared to intramuscular injection. In contrast, intranasal administration of 1 mg / kg doses of epinephrine compositions containing C12E23 and C16E10, respectively, resulted in significantly reduced activity, indicating some degree of adverse reaction. Following intranasal administration of an adrenaline combination containing C12E8, C12E4, or C10E6 (1 mg / kg dose of adrenaline), subjects remained normal and active, and higher F values ​​and shorter T values ​​were observed. max And a larger area under the curve (AUC) in the early fraction of the drug. These observations indicate that C12E7, C12E8, C12E4, and C10E6 unexpectedly improve tolerance to epinephrine. When C12E7, C12E8, C12E4, and C10E6 are added to an epinephrine composition, the subject's tolerance to the composition is improved. In some embodiments, the epinephrine composition is administered to the subject via nasal administration.

[0090] In the context of this invention, a test subject exhibiting better "tolerance" to a drug or pharmaceutical composition is defined as a test subject that, after administration of a composition containing the drug or pharmaceutical composition (such as adrenaline or an adrenaline composition), exhibits fewer adverse reactions, milder adverse reactions, or is less likely to experience adverse reactions. Tolerance describes the test subject's ability to tolerate high doses of an active ingredient (e.g., adrenaline).

[0091] As used herein, irreversible damage refers to damage that cannot spontaneously recover within a relatively short period of time. Due to the potent local pharmacological effects of adrenaline, irreversible damage may be exacerbated when permeation enhancers are used in conjunction with adrenaline. For the purposes of this invention, an insulin absorption test is used to assess nasal mucosal damage and determine whether the damage is irreversible (see Arnold, John J., et al., "Reestablishment of the nasal permeability barrier to several peptides following exposure to the absorption enhancer tetradecyl-β-D-maltoside," Journal of Pharmaceutical Sciences, 99.4 (2010): 1912-1920). Under basal conditions, the nasal epithelium severely restricts the absorption of drugs with molecular weights greater than 1 kDa. Therefore, insulin with molecular weights greater than 5 kDa generally cannot penetrate the nasal mucosa unless the nasal mucosa is damaged. Insulin administered nasally after exposure to a permeation enhancer can be monitored by monitoring the C-value of insulin administered nasally at different time points following nasal administration of a drug-containing (i.e., adrenaline) composition. max The AUC was used to determine whether the composition caused any damage to the nasal mucosa of the test subject, and whether such damage (if any) was reversible. Lower levels of insulin exposure and C... max The AUC indicates less damage to the nasal mucosa, while higher levels of insulin exposure result in lower C. max The AUC indicates damage to the nasal mucosa. In cases of reversible nasal mucosal damage, insulin C... max The AUC initially appeared high, then rapidly declined, returning to a significantly lower level within approximately 2 hours. In cases of irreversible nasal mucosal damage, insulin's C... max The AUC will remain at a high level for more than 2 hours. See Example 5 for a detailed protocol for the insulin absorption test.

[0092] This invention also discloses an unexpected finding: the pharmaceutical compositions exhibit faster reversal of nasal mucosal damage under low pH conditions (pH below 7, for example, in the range of 4-6). When the pharmaceutical compositions are formulated to a pH below 7, they induce only reversible damage that recovers more quickly. In contrast, many penetration enhancers known in the art and used in intranasal formulations, such as DDM or DEGEE, do not exhibit the pH-sensitive trend in nasal mucosal damage disclosed in Example 5.

[0093] Furthermore, the pharmaceutical composition exhibits better stability and permeability under low pH conditions, i.e., pH values ​​below 7, for example, in the range of 4-6.

[0094] Compared to previously disclosed compositions (such as those containing penetration enhancers like C12E9, DDM, and DEGEE), it has been unexpectedly found that the current compositions exhibit less nasal mucosal damage under low pH conditions, i.e., pH values ​​below 7, for example, in the range of 4–6. For example, it has been reported that C12E9 causes severe nasal mucosal damage when administered nasally in a composition at pH 7.4 (Bleske et al., Effect of Vehicle on the Nasal Absorption of Epinephrine during Cardiopulmonary Resuscitation, Pharmacotherapy, 16(6), 1039–1045 (1996)). However, this invention discloses a finding that C12E9 causes significantly less nasal mucosal damage under low pH conditions, i.e., pH values ​​below 7, for example, in the range of 4–6.

[0095] In some embodiments, the pH of the pharmaceutical composition is below 7. In some embodiments, the pH range of the pharmaceutical composition is from 4 to 6. In some embodiments, the pH of the pharmaceutical composition is 4, 5, or 6.

[0096] The present invention also discloses a pharmaceutical composition comprising adrenaline and C12E9 as a penetration enhancer, wherein the pH of the pharmaceutical composition is below 7, for example, in the range of 4 to 6.

[0097] stability

[0098] In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is a pH adjuster, antioxidant, preservative, or osmotic pressure regulator.

[0099] pH adjusters, sometimes also called pH modifiers or acidity regulators, are substances used to adjust the pH of a pharmaceutical composition to a specific range. pH is a way of expressing the concentration of hydrogen ions in water. Specifically, pH is the concentration of hydrogen ions (H+) in an aqueous solution. + The negative logarithm of the concentration (mol / L): pH = -log 10 (H + In some implementations, the pH adjuster is an acid or a base. Some examples of pH adjusters include hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, and ammonia.

[0100] Antioxidants are compounds that inhibit oxidation. In some embodiments, antioxidants are used to improve the stability of a pharmaceutical composition by delaying the oxidation of the active substance and other excipients. Some examples of antioxidant excipients include cysteine ​​(CYS), sodium metabisulfite (SMB), propyl gallate (PG), butylated hydroxytoluene (BHT), D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS), ascorbic acid (VC), methionine, sodium bisulfite, sodium sulfite, alpha-lipoic acid, and butylated hydroxyanisole (BHA) (Celestino et al., Brazilian J. Pharma. Sci. 43-3, 405-415 (2012)).

[0101] Preservatives are substances added to a pharmaceutical composition to prevent adverse physical, chemical, or biological changes. In some embodiments, preservatives are bactericides or antimicrobial agents. Some examples of preservatives include benzalkonium chloride, chlorobutanol, butylparaben, propylparaben, benzyl chloride, chlorocresol, phenol, and benzoic acid.

[0102] Osmotic regulators are typically low-molecular-weight, water-soluble substances. Osmosis is the diffusion of water across a membrane under osmotic pressure caused by an imbalance of molecules on either side. Osmotic regulators can alter the osmotic pressure across cell membranes. Some examples of osmotic regulators include sodium chloride, glucose, mannitol, sorbitol, lactose, phosphoric acid, and citric acid.

[0103] In some embodiments, the pharmaceutical composition comprises an antioxidant. In some embodiments, the antioxidant is selected from sodium bisulfite, sodium metabisulfite (SMB), propyl gallate (PG), sodium sulfite, ascorbic acid (VC), methionine, alpha-lipoic acid, cysteine ​​(CYS), D-α-tocopherol polyethylene glycol succinate (vitamin E TPGS), butylated hydroxytoluene (BHT), and butylated hydroxyanisole (BHA). In some embodiments, the antioxidant is selected from sodium bisulfite and sodium metabisulfite. In some embodiments, the pharmaceutical composition comprises 0.2% sodium bisulfite. As shown in Example 4, when sodium bisulfite (as an antioxidant) is used in conjunction with EDTA-2Na (as a chelating agent), the adrenaline formulation containing C10E6, C12E4, C12E7, C12E8, and C12E9 as penetration enhancers exhibits significantly better color stability characteristics. Pharmaceutical compositions containing penetration enhancers and antioxidants exhibit better stability and permeability under low pH conditions, i.e., pH values ​​below 7, such as the range of 4-6.

[0104] As used herein, "stability," in the context of a pharmaceutical composition, refers to the ability of a pharmaceutical composition to retain its chemical, physical, and biopharmaceutical properties over time. For the purposes of this disclosure, stability is expressed as a "percentage of content" of the active ingredient in the pharmaceutical composition. As used herein, the percentage of content of the active ingredient on day n is calculated by dividing the amount of the active ingredient on day n by the amount of the active ingredient on day 0, wherein the composition is stored under given environmental conditions (pH, temperature) from day 0 throughout the accelerated stability test, and the percentage of content is a weight percentage.

[0105] In some embodiments, the pharmaceutical composition, at pH 4.0 and 60°C, has a content percentage of not less than 90% on day 30.

[0106] method

[0107] On the other hand, the present invention discloses a method comprising administering an effective amount of the pharmaceutical composition to a test subject. Routes of administration may include, for example, intravenous, intratumoral, oral, rectal, vaginal, parenteral, local, pulmonary, intranasal, buccal, and ocular administration.

[0108] In some implementations, this method is used to treat subjects suffering from type I hypersensitivity reactions. Type I hypersensitivity reactions are IgE-mediated immune responses involving the release of antibodies against soluble antigens. This leads to mast cell degranulation and the release of histamine and other inflammatory mediators. Type I hypersensitivity reactions include atopic diseases, which are excessive IgE-mediated immune responses (e.g., rhinitis, conjunctivitis, and dermatitis); and allergic diseases, which are immune responses to exogenous allergens (e.g., anaphylactic shock, urticaria, angioedema, food allergies, and drug allergies).

[0109] In some implementations, the allergic reaction is selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, anaphylactic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

[0110] In some embodiments, the pharmaceutical composition is administered to the test subject via nasal delivery.

[0111] As used in this article, the term "C" max "C" refers to the maximum value of the blood concentration shown on a curve representing the change in blood concentration of the active pharmaceutical ingredient (e.g., adrenaline) or its metabolites over time. It is understood that those skilled in the art can select appropriate methods and conditions to measure the blood concentration of adrenaline in a subject and determine C. max It is understood that those skilled in the art can select appropriate methods and conditions to measure the blood concentration of adrenaline in a subject and determine the Tg. max .

[0112] As used herein, AUC refers to the area under the plasma concentration-time curve, which is the area under the curve showing the change in plasma concentration of the active ingredient (e.g., adrenaline) over time after administration of a given dose of the active ingredient (e.g., epinephrine). "AUC" 0-∞ "AUC" is the area under the concentration-time curve extrapolated to infinity after drug administration. 0-t " is the area under the concentration-time curve from time 0 to time t after drug administration, where t is the last time point at which the concentration can be measured.

[0113] As used herein, bioavailability (F) describes the percentage of a drug dose that reaches systemic circulation. Its meaning is as defined in 21 C.FR § 320.1(a). For the purposes of this disclosure, F specifically refers to the ratio of exposure per unit dose between intranasal administration and intramuscular injection. For example, F(AUC) 0-t ) = (AUC 0-t , 鼻内 / dose 鼻内 ) / (AUC 0-t , 肌肉注射 / dose 肌肉注射 )×100%.

[0114] As used in this article, early partial bioavailability (F) 0-10 Specifically, it refers to the ratio of AUC within the first 10 minutes after administration via intranasal or intramuscular injection. For example, F(AUC) 0-10 ) = (AUC 0-10 , 鼻内 / dose 鼻内 ) / (AUC 0-10 , 肌肉注射 / dose 肌肉注射 )×100%. Example

[0115] This specification can be further described by the following non-limiting embodiments, wherein standard techniques known to those skilled in the art, as well as techniques similar to those described in these embodiments, may be used where appropriate. It will be understood that those skilled in the art will contemplate other embodiments consistent with the description provided herein.

[0116] Example 1: Evaluation of PAMPA in vitro permeability

[0117] The effects of different permeation enhancers on the in vitro permeability of adrenaline were evaluated using a 96-well PAMPA Sandwich set (PION Inc., MA, USA).

[0118] PRISMA TM The buffer solution was prepared by the following method: 25 mL of PRISMA... TM (P / N110151, PION Inc., MA, USA) Dilute to 1 L with ultrapure water, then adjust pH to 4.0 with 0.5 M NaOH solution.

[0119] Add 200 μL of hydration solution (PION Inc., MA, USA) to each well of the supply end (P / N 110660, PION Inc., MA, USA). Then, immerse the pre-coated PAMPA plate (P / N120657, PION Inc., MA, USA) in the hydration solution overnight for full hydration.

[0120] The designed compositions were prepared by dissolving osmosis enhancers (concentrations listed in Table 2) together with epinephrine (10 mg / mL, 50 mg / mL, 3 mg / mL) in an aqueous buffer at pH 4.0. An aqueous epinephrine solution at pH 4.0 (without osmosis enhancers) served as the experimental control. 200 μL of each composition solution was accurately added to each well of the supply end, followed by 200 μL of LPRISMA. TM Buffer solution was added to each well of the receiver. The PAMPA artificial membrane was then placed between the supply and receiver plates to initiate the in vitro permeability evaluation experiment. Permeability tests for each composition were performed in quadruplicate. The PAMPA kit, including the supply, PAMPA artificial membrane, and receiver, was capped and incubated at 37°C for 5 hours. Antioxidants were also used to prevent oxidation during the experiment. After incubation, the epinephrine concentration in the supply and receiver solutions was determined by UPLC using the methods described in Table 1.

[0121] Table 1. UPLC analysis method for assessing adrenaline permeability in vitro using the PAMPA assay

[0122]

[0123] Polyoxyethylene alkyl ethers containing double bonds in the alkyl chain are written as C18-1E10 and C18-1E20. Table 2 lists the PAMPA test results of epinephrine solutions containing different types of penetration enhancers (0.1%, 0.25%, 2.5% w / v or v / v). Compared with the negative control (epinephrine solution without penetration enhancer, only 1.1% penetrated to the receiver, Pe value 0.45 × 10⁻⁶), the negative control showed significantly lower penetration. -6Compared to solutions containing 0.1%–2.5% C10E6, C12E7, C12E8, C12E9, and C16E10 (cm / s), epinephrine solutions consistently showed significantly higher epinephrine concentrations in the receiver solution and higher Pe values, indicating that over 8% of the epinephrine permeated to the receiver and the Pe value was higher than 3 × 10⁻⁶. -6 cm / s. Compared with DEGEE, all PAMPA in vitro permeability tests showed significantly increased permeability in compositions containing 0.1%–2.5% of C10E6, C12E7, C12E8, C12E9, and C16E10; some permeation enhancers exhibited higher permeability than DDM, especially at lower permeation enhancer concentrations, which is more clinically significant.

[0124] Table 2. Results of in vitro permeability tests of adrenaline solutions containing different types of permeability enhancers

[0125]

[0126]

[0127] Example 2: Pharmacokinetic (PK) Study of Compositions Containing Permeability Enhancers After Nasal Administration. The preparation methods for different formulation compositions for intranasal administration are as follows: First, an aqueous solution containing 0.25% permeability enhancer, 0.2% sodium bisulfite, and 0.9% sodium chloride was prepared. Then, adrenaline was dissolved to achieve a concentration of 10 mg / mL or 3 mg / mL. All formulation solutions were then adjusted to pH 4 and refrigerated. The liquid compositions containing adrenaline and permeability enhancers were administered intranasally according to the methods described in Table 3.

[0128] The prepared solution composition was administered intranasally to Sprague-Dawley (SD) rats. Blood samples were then collected into 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90, and 120 minutes after administration. A 10% (w / v) sodium metabisulfite aqueous solution was prepared and added to the sample at a volume ratio of 9:1 to prevent oxidation. The sample was vortexed thoroughly and stored in an ice-water bath pending pretreatment. If sample pretreatment and analysis were not performed on the day of the PK experiment, the sample was stored at -80°C. For frozen samples, equilibration was performed at room temperature before further pretreatment. After the sample thawed, 20 μL of plasma sample was transferred to a 1.5 mL polyethylene centrifuge tube, and 180 μL of internal standard solution (preparation method shown in Table 5) was added. The mixed sample was vortexed for 5 minutes and centrifuged at 10,000 rpm and 4°C for 5 minutes. Transfer 130 μL of supernatant to a 96-well plate, add 130 μL of ultrapure water, vortex for 5 minutes, and centrifuge at 4,000 rpm and 4 °C for 5 minutes. The pretreated samples were analyzed by LC-MS / MS, and the methods are listed in Tables 4, 6, and 7.

[0129] Table 3. Experimental design for rat pharmacokinetic studies of different formulation compositions

[0130]

[0131] Table 4. HPLC Analysis Methods for Adrenaline

[0132]

[0133]

[0134] Table 5. Preparation method of internal standard solution

[0135]

[0136] Table 6. Methods for Adrenal Serum Spectroscopy Analysis

[0137]

[0138] Table 7. Methods for Adrenal Gland Plasma Spectroscopy Analysis

[0139]

[0140] The changes in plasma adrenaline concentration over time in the 10 mg / mL epinephrine prescription group (dosage dose 1 mg / kg) are shown in Table 8. Figure 1As shown. In the intramuscular injection PK study, each group used 3 SD rats, with an adrenaline concentration of 1 mg / mL and a dose of 0.1 mg / kg. In the nasal administration PK study, each group used 2 SD rats, with an adrenaline concentration of 10 mg / mL and a dose of 1 mg / kg. In Examples 2 and 3, "NA" indicates that adrenaline was not detected in plasma at a specific time point or that the parameter could not be calculated due to the lack of adrenaline detection at that time point. PK parameters, i.e., T, were calculated using MaS Studio (v1.5.3.10). max C max AUC, bioavailability (F 0-t ) and early partial bioavailability (F) within the first 10 minutes after administration 0-10 As shown in Table 9. Observations revealed that when polyoxyethylene alkyl ethers have 9-15 methylene groups and 4-10 oxyethylene units, C max The AUC and bioavailability were significantly higher.

[0141] Table 8. Changes in plasma adrenaline concentration over time after intranasal administration of drug delivery compositions containing different penetration enhancers.

[0142]

[0143]

[0144]

[0145] Table 9. PK parameters calculated based on the data listed in Table 8 (samples marked with * are analyzed using the method in Table 5, samples ...8, samples marked with * are analyzed using the method in Table 5). The samples were analyzed using the methods shown in Table 6.

[0146]

[0147] The changes in plasma adrenaline concentration over time in the 3 mg / mL epinephrine prescription group (far lower than the clinically used 0.3 mg / kg dose) are shown in Table 10. Figure 2 As shown. The PK parameters, i.e., T, are calculated using MaS Studio (v1.5.3.10). max C max The AUC and bioavailability are shown in Table 11. Observations revealed that when polyoxyethylene alkyl ethers have 9-15 methylene groups and 4-10 oxyethylene units, T... max Shorter, while maintaining a relatively high C max AUC and bioavailability. From a clinical perspective, a shorter T... max Even more ideally, because for treating type I hypersensitivity reactions, adrenaline needs to work more quickly.

[0148] Table 10. Changes in plasma adrenaline concentration over time in nasal administration compositions containing different types of penetration enhancers.

[0149]

[0150] Table 11. PK parameters calculated based on the data listed in Table 10 (for samples marked with *, the parameters are calculated using the PK parameters ...

[0151] Table 5 shows the analysis methods and the analysis of items with... The samples were analyzed using the methods shown in Table 6.

[0152]

[0153] Example 3: Pharmacokinetic (PK) study of compositions with different concentrations of penetration enhancer and adrenaline.

[0154] The preparation process of the intranasal administration composition is as follows: First, prepare an aqueous solution containing 0.1%, 0.25%, 1.0%, and 2.5% penetration enhancer, 0.2% sodium bisulfite, and 0.9% sodium chloride. Then, dissolve epinephrine to achieve a drug concentration of 10 mg / mL or 3 mg / mL (except for the 2.5% DEGEE composition, whose epinephrine concentration is 25 mg / mL); adjust the prescription solution to pH 4 and refrigerate. The liquid composition containing epinephrine and penetration enhancer is administered intranasally according to the method in Table 12.

[0155] Different formulations of the composition were instilled into the nasal cavity of SD rats. Blood samples were collected into 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90, and 120 minutes after administration. A 10% (w / v) sodium metabisulfite aqueous solution was prepared and added to the sample at a volume ratio of 9:1 to prevent oxidation. The sample was vortexed thoroughly and stored in an ice-water bath pending pretreatment. If sample pretreatment and analysis were not performed on the day of the PK experiment, the sample was stored at -80°C. For frozen samples, equilibration was performed at room temperature before further pretreatment. After the sample thawed, 20 μL of plasma sample was transferred to a 1.5 mL polyethylene centrifuge tube, and 180 μL of internal standard solution (preparation method shown in Table 5) was added. The mixed sample was vortexed for 5 minutes and centrifuged at 10,000 rpm and 4°C for 5 minutes. Transfer 130 μL of supernatant to a 96-well plate, add 130 μL of ultrapure water, vortex for 5 minutes, and centrifuge at 4,000 rpm and 4 °C for 5 minutes. The pretreated samples were analyzed by LC-MS / MS, and the methods are listed in Tables 4, 6, and 7.

[0156] Table 12. Experimental Design of Rat PK Study

[0157]

[0158] Table 13 shows the plasma adrenaline concentrations at different time points after administration of compositions with different adrenaline concentrations, types and concentrations of osmosis enhancers. Figure 3 It was observed that the tested polyoxyethylene alkyl ethers exhibited significant permeation-enhancing capabilities within a concentration range of 0.1 mg / mL to 2.5 mg / mL. The PK parameters, i.e., T, were calculated using MaS Studio (v1.5.3.10). max C max AUC, F 0-t and F 0-10 See Table 14. It was observed that, at a constant administered volume, C increased with increasing concentrations of the osmotic enhancer and adrenaline. max AUC and bioavailability are typically increased. Compared to groups containing DDM and DEGEE, polyoxyethylene alkyl ethers exhibited higher C values ​​at the same penetration enhancer and epinephrine concentrations. max The AUC and bioavailability suggest superior penetration-enhancing ability. For formulations containing polyoxyethylene alkyl ethers, a shorter T0 was also observed. max This indicates that adrenaline takes effect faster.

[0159] Table 13. Changes in plasma adrenaline concentration over time after administration of compositions with different adrenaline concentrations, types and concentrations of osmotic enhancers.

[0160]

[0161]

[0162] Table 14. PK parameters calculated based on the data listed in Table 13

[0163]

[0164]

[0165] Example 4: Pharmacokinetic (PK) Study of Compositions Containing Permeability Enhancers After Nasal Administration. The preparation methods for different intranasal formulations are as follows: First, an aqueous solution containing 0.25% permeability enhancer, 0.2% sodium metabisulfite, and 0.9% sodium chloride was prepared. Then, adrenaline was dissolved to achieve adrenaline concentrations of 2.5 mg / mL, 10 mg / mL, or 20 mg / mL. All formulation solutions were then adjusted to pH 4 and refrigerated. The liquid compositions containing adrenaline and permeability enhancers were administered into the nasal cavity of beagle dogs according to the methods described in Table 15.

[0166] The composition was administered intranasally to beagle dogs. Blood samples were collected at 1, 5, 10, 15, 20, 30, 60, 90, and 120 minutes after administration. Whole blood samples were collected into anticoagulant tubes containing EDTA-K2 and centrifuged at 1524 g and 4°C for 10 minutes. Aliquots were then collected and stored at -40°C to -20°C for analysis. The samples to be analyzed were thawed and vortexed at room temperature. 100 μL of the sample was taken, and 20 μL of water (containing 0.1% acetic acid and 50 ng / mL adrenaline-D6) was added as a protein settling agent. Then, 250 μL of PBA was added, and the mixture was vortexed. 400 μL of TOAB was added, and the mixture was vortexed for 10 minutes, followed by centrifugation at 13000 rpm for 5 minutes. Transfer 300 μL of the supernatant to a 1.5 mL centrifuge tube, add 200 μL of n-octanol and 125 μL of 0.02 N hydrochloric acid aqueous solution, and vortex for 3 min. Collect the lower layer solution and mix it with 50 μL of sodium tetraborate buffer (100 mmol) and 100 μL of benzoyl chloride (1%), and vortex for 0.5 min. Inject 10 μL of the final mixed sample solution into the LC-MS / MS system for analysis according to the methods listed in Tables 16 and 7.

[0167] Table 15. Experimental Design of Beagle PK Studies with Different Formula Compositions

[0168]

[0169]

[0170] Table 16. HPLC Analysis Methods for Adrenaline

[0171]

[0172] Table 17 and Figure 4 This study shows the changes in plasma adrenaline concentrations over time at different time points after administration for formulations with different dosage concentrations and types of penetration enhancers. The corresponding pharmacokinetic parameters (i.e., T0) are also presented. max C max AUC, bioavailability relative to intramuscular injection F 0-t Early partial bioavailability F within the first 10 minutes after administration 0-10 The results were calculated using Phoenix WinNonlin 8.3 software and are shown in Table 18. Compared to commercial formulations using DDM as a penetration enhancer, the C12E7 group showed higher bioavailability even at lower epinephrine concentrations and dosages. The C12E7 group also showed a nearly 3-fold higher early partial AUC compared to the DDM group (the currently used commercial formulation). Notably, a higher early partial AUC is highly desirable when using epinephrine in emergency situations to treat type I anaphylactic reactions (including anaphylactic shock).

[0173] Table 17. Changes in plasma adrenaline concentration over time after intranasal administration of compositions containing different penetration enhancers.

[0174]

[0175] Table 18. PK parameters calculated based on the data listed in Table 17

[0176]

[0177] Example 5: Stability Evaluation of the Composition

[0178] A nasal composition with an adrenaline concentration of 10 mg / mL was prepared by dissolving adrenaline in aqueous solutions containing 0.25% of different permeation enhancers and 0.9% sodium chloride. To prepare formulations containing antioxidants and preservatives, adrenaline was dissolved in aqueous solutions containing 0.1%–1.0% of different permeation enhancers, 0.2% of antioxidants, 0.1% of preservatives, and 0.9% sodium chloride. To prepare formulations containing antioxidants under different pH conditions, adrenaline was dissolved in aqueous solutions containing 1.0% of different permeation enhancers, 0.2% of antioxidants, and 0.9% sodium chloride, with the pH adjusted to 4.0, 5.0, 6.0, and 7.0, respectively. For the composition containing 1.0% C12E9, a sample at pH 7.4 was also prepared, containing 0.2% antioxidant and phosphate-buffered saline (PBS). The stability test design for the compositions is shown in Table 19.

[0179] Table 19. Design of Formulation Stability Tests

[0180]

[0181] The liquid composition was dispensed into vials, the caps were then sealed, and the vials were placed in an oven for accelerated stability testing. This was done according to the United States Pharmacopeia method (USP 35). <391> EPINEPHRINE ASSAY was used to quantify the adrenaline content in different compositions before and after accelerated stability testing using high-performance liquid chromatography (HPLC). The analytical methods are shown in Table 20. The phosphate buffer (pH 2.8) was prepared as follows: 5.0 g / L potassium dihydrogen phosphate and 2.6 g / L sodium octane sulfonate were added to water, and the pH was adjusted to 2.8. The phosphate buffer was filtered through a 0.45 μm filter before use.

[0182] Table 20. HPLC Analysis Methods for Adrenaline

[0183]

[0184]

[0185] Table 21 shows the stability test results of compositions containing adrenaline and different penetration enhancers after being accelerated at 60°C for different times. It was observed that the adrenaline content of all formulations decreased significantly after acceleration. C12E9 and C16E2 exhibited poorer stability characteristics compared to the other compositions. The remaining compositions showed comparable stability characteristics.

[0186] Table 21. Changes in adrenaline content (%) in compositions containing adrenaline and different osmosis enhancers under accelerated conditions at 60°C.

[0187]

[0188] The stability test results (accelerated at 60°C for different times) of compositions containing 10 mg / mL adrenaline, 0.25% C12E8 or DDM, antioxidants, chelating agents, and preservatives are shown in Table 22. It was observed that sodium bisulfite significantly improved the stability of the compositions; when sodium bisulfite and EDTA-2Na were used simultaneously, the composition remained clear for a longer period. Compositions without sodium bisulfite turned red to dark brown after acceleration.

[0189] Table 22. Changes in adrenaline content (%) after accelerated treatment at 60°C with antioxidants, chelating agents, and preservatives.

[0190]

[0191] The stability test results of compositions containing 10 mg / mL adrenaline, 0.1% or 1.0% of different penetration enhancers, 0.2% of different antioxidants, and 0.1% of different preservatives after accelerated testing at 60°C are shown in Tables 23 and 24. No significant difference in stability was observed when two preservatives were added simultaneously, indicating good chemical compatibility.

[0192] Table 23. Changes in adrenaline content (%) in compositions containing different penetration enhancers and preservatives (all compositions contain sodium bisulfite, accelerated at 60°C).

[0193]

[0194] Table 24. Changes in adrenaline content (%) of compositions containing different penetration enhancers and preservatives (all compositions contain sodium metabisulfite, accelerated at 60°C).

[0195]

[0196] Table 25 shows the stability test results of the composition containing 10 mg / mL epinephrine, 1.0% of different penetration enhancers, and 0.2% sodium bisulfite at pH 4.0, 5.0, 6.0, and 7.0 after accelerated treatment at 60°C. It was observed that the antioxidant sodium bisulfite significantly improved the stability of the composition within the pH range of 4.0 to 6.0, with the epinephrine content still above 95% after 7 days of accelerated treatment at 60°C. However, the stability of the composition decreased rapidly when the pH was equal to or higher than 7.0. Furthermore, it was observed that sodium bisulfite only improved the stability of the composition within the pH range of 4.0 and 6.0; when the pH was equal to or higher than 7.0, the addition of sodium bisulfite resulted in worse stability of the composition.

[0197] Table 25. Changes in adrenaline content (%) after acceleration at 60°C for different compositions

[0198]

[0199]

[0200] Example 6: Evaluation of nasal mucosal damage after intranasal administration of an adrenaline-containing composition

[0201] Since both permeation enhancers and epinephrine can cause nasal mucosal damage, experiments were designed and conducted to evaluate nasal mucosal damage following nasal administration of compositions containing epinephrine and different permeation enhancers. Human insulin is almost impossible to absorb through the nasal mucosa unless it is damaged. Therefore, as described in the literature (see Arnold, John J., et al., "Reestablishment of the nasal permeability barrier to several peptides following exposure to the absorption enhancer tetradecyl-β-D-maltoside," Journal of Pharmaceutical Sciences, 99.4(2010):1912-1920), nasal mucosal damage was assessed by the absorption of insulin through the nasal mucosa at different time points after nasal administration of compositions containing permeation enhancers and epinephrine. Significant nasal insulin absorption indicated nasal mucosal damage, while no insulin absorption indicated an intact nasal mucosa.

[0202] The evaluation method is as follows: First, a liquid composition containing a penetration enhancer and adrenaline was prepared according to Table 26, and a 0.5 μU / mL conventional human insulin solution was also prepared; 8 healthy SD rats were used in each composition test group, and 2 SD rats were used in each time point subgroup of each composition; then, each composition was administered intranasally (dose 0.1 mL / kg) to rats in all the corresponding 4 time point subgroups as listed in Table 22. For the 0-hour subgroup, conventional human insulin (0.05 μU / kg, 0.5 μU / mL, 0.1 mL / kg) was administered intranasally to SD rats immediately after administration of the epinephrine composition. For the 2-hour subgroup, conventional human insulin (0.05 μU / kg, 0.5 μU / mL, 0.1 mL / kg) was administered intranasally to SD rats 2 hours after administration of the epinephrine composition. For the 4-hour subgroup, conventional human insulin (0.05 μU / kg, 0.5 μU / mL, 0.1 mL / kg) was administered intranasally to SD rats 4 hours after administration of the epinephrine composition. For the 8-hour subgroup, conventional human insulin (0.05 μU / kg, 0.5 μU / mL, 0.1 mL / kg) was administered intranasally to SD rats 8 hours after administration of the epinephrine composition. Blood samples were collected from all rats at 0, 10, 20, 30, 45, 60, 90, and 120 minutes after administration of conventional human insulin. Plasma insulin concentrations were then measured using an ELISA kit (Mercodia brand, operated according to instructions). PK parameters for each subgroup were calculated using MaS Studio software (v1.5.3.10) (Table 27). Figure 5 and Figure 6 This allows us to reflect the changes in nasal mucosal damage over time after administration of different compositions.

[0203] Table 26. Experimental design for changes in nasal cavity permeability

[0204]

[0205]

[0206]

[0207]

[0208] Surprisingly, the pharmacokinetic parameters of insulin showed that compositions containing C12E9 exhibited irreversible damage at higher pH values ​​(e.g., pH 7.4), while milder and reversible damage was observed at lower pH values ​​(e.g., pH 4.0). Compared to the irreversible nasal mucosal damage observed with DDM, DEGEE, and C12E9 (high pH), all compositions containing polyoxyethylene alkyl ethers disclosed in this application showed a faster recovery of the nasal mucosa to normal (within 2 hours) at lower pH values. Furthermore, higher insulin AUC and C2E9 were observed at 0 hours for the tested compositions containing polyoxyethylene alkyl ethers. max This indicates that drug absorption is faster compared to DDM, DEGEE, and C12E9 (high pH). Therefore, especially when used at an appropriate pH, the polyoxyethylene alkyl ether disclosed in this application can serve as a safer (e.g., faster nasal mucosal recovery) and more effective (e.g., faster drug absorption after administration) penetration enhancer compared to other penetration enhancers disclosed in the prior art.

[0209] Table 27. Reversibility of the permeation barrier of nasal insulin by different permeation enhancers

[0210]

[0211]

Claims

1. A pharmaceutical composition comprising adrenaline and a permeation enhancer having the following formula: CH3(CH2) n-1 [OCH2CH2] m OH, wherein n is an integer selected from 10, 12 and 16, m is an integer selected from 4, 5, 6, 7 and 8, the pH value of the pharmaceutical composition is in the range of 4 to 6, and the pharmaceutical composition is for nasal administration or for use in a nasal administration device.

2. The pharmaceutical composition according to claim 1, wherein n is 12 and m is selected from 4, 7 and 8.

3. The pharmaceutical composition according to claim 1, wherein n is 10 and m is 6.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of the penetration enhancer ranges from 0.1% to 2.50% (v / v).

5. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of the penetration enhancer is 0.25% (v / v).

6. The pharmaceutical composition according to any one of claims 1 to 3 further comprises at least one pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is a pH adjuster, antioxidant, preservative or osmotic pressure regulator.

7. The pharmaceutical composition according to claim 6, wherein the antioxidant is selected from sodium bisulfite, sodium metabisulfite (SMB), propyl gallate (PG), sodium sulfite, ascorbic acid (VC), methionine, alpha-lipoic acid, cysteine ​​(CYS), D-α-tocopherol polyethylene glycol succinate (vitamin ETPGS), butylated hydroxytoluene (BHT), and butylated hydroxyanisole (BHA).

8. The pharmaceutical composition according to claim 6, wherein the antioxidant is selected from sodium bisulfite and sodium metabisulfite.

9. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of adrenaline in the pharmaceutical composition ranges from 0.3% to 5% (w / v).

10. The pharmaceutical composition according to any one of claims 1 to 3, wherein the dosage form of the pharmaceutical composition is a liquid or a spray.

11. The pharmaceutical composition according to any one of claims 1 to 3, wherein the recovery rate of the pharmaceutical composition on day 30 is not less than 90% by weight under conditions of pH 4.0 and 60°C.

12. The pharmaceutical composition according to any one of claims 1 to 3, wherein the absolute value of Pe in the PAMPA test is greater than 3 × 10⁻⁶. -6 cm / s.

13. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition does not cause irreversible damage to the nasal mucosa.

14. Use of the pharmaceutical composition according to any one of claims 1 to 13 in the preparation of a medicament for treating an allergic reaction in a test subject.

15. The use according to claim 14, wherein the drug is used to treat a subject suffering from a type I hypersensitivity reaction.

16. The use according to claim 14, wherein the drug is used to treat conditions selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, anaphylactic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

17. The use according to any one of claims 14 to 16, wherein the pharmaceutical composition is administered to the test subject via nasal administration.

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