Novel formulations of epinephrine and uses thereof

Abstract

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

Description

Field of the Invention

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

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

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

[0004] background

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

[0006] Specifically, epinephrine is the drug of choice for treating type I allergic reactions, including anaphylactic shock. It is estimated that up to 2% of the world's population will experience anaphylaxis at some point in their lives, and the trend is increasing (Simons et al., The World Allergy Organization Journal, 4 (2): 13–37 (2006)). Anaphylactic shock is a severe, potentially fatal allergic reaction and medical emergency that develops rapidly and requires immediate medical attention, regardless of whether emergency medications are available 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 epinephrine autoinjector was developed to more rapidly self-administer epinephrine via intramuscular injection in emergency situations (Mylan Specialty LP " -Epinephrine injection, EPIPEN -epinephrine injection" (FDA product label (archived on February 1, 2014 (PDF), retrieved on January 22, 2014). However, epinephrine auto-injectors have a high probability of causing dangerous consequences, such as misuse for subcutaneous or intravenous injections, or the use of the wrong dose (Bilò, M. Beatrice., Anaphylaxis caused by Hymenoptera stings: from epidemiology to treatment, Allergy 66, pages 35-37 (2011)). Epinephrine injectors are also structurally complex, which can lead to frequent mechanical failures, making this life-saving drug less reliable than it should be. In addition, patients (whether or not they suffer from needle phobia) are reluctant to use auto-injectors in public places and often postpone their use until the disease is serious, thus delaying treatment and ultimately failing to effectively reverse the rapid progression of type I anaphylaxis.

[0007] In view of the challenges faced by epinephrine auto-injectors, there is an urgent need for a more reliable and patient-compliant way to deliver epinephrine. Nasal administration is one of the promising routes of administration, and there are currently multiple epinephrine nasal spray preparations under development. However, existing epinephrine nasal spray preparations are plagued by low absorption, nasal mucosal damage, and poor chemical stability. For example, compared with intramuscular injection, even with the use of a penetration enhancer, existing epinephrine nasal spray preparations still require more than three times the dose (0.3mg vs 1mg) (Australian Patent No. AU2019217643B2). It is well known that the therapeutic window of epinephrine is very narrow and poorly absorbed, which is often associated with large differences in absorption rates. Therefore, when a higher dose is used to solve the problem of poor absorption, for those patients with higher absorption rates, drug overdose may result, thereby causing serious adverse reactions, such as cerebral hemorrhage, hemiplegia, subarachnoid hemorrhage, dyspnea, etc. A similar situation has been identified as a major safety risk for another drug that requires further improvement (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). In addition, the European Medicines Agency (EMA) clearly stated that the early partial AUC value, especially the AUC in the first 10 minutes after administration, is crucial to the efficacy of epinephrine nasal spray; Tmax There is also a correlation. At the same epinephrine dose, T max Should be equivalent to intramuscular (IM) or subcutaneous (SC) injection; C max and total AUC are considered to be the most relevant parameters for drug safety and efficacy. Neffy is an adrenaline nasal spray for which a new drug application (NDA) was submitted to the EMA, but was not approved due to a smaller AUC in the early portion compared with intramuscular (IM) injection (Assessment Report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0008] In addition, it has been reported that the use of penetration enhancers and epinephrine in nasal spray preparations can cause 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 very easy to be oxidized, so it is very challenging to maintain the stability of epinephrine preparations (GB West, Oxidation of Adrenaline in Alkaline Solution, British Journal of Pharmacology and Chemotherapy, Volume 2, Issue 2, p. 121-130, 1947). Therefore, it is necessary to develop an epinephrine nasal spray preparation with higher bioavailability, less nasal mucosal damage, less individual variability, faster absorption rate, and better chemical stability. Summary of the invention

[0009] The invention discloses a novel adrenaline preparation containing polyoxyethylene alkyl ether as a penetration enhancer, a method for administering the adrenaline preparation, and its use. The preparation disclosed in the invention can have higher bioavailability, faster absorption rate, better chemical stability, and less damage to the nasal mucosa.

[0010] In one aspect, the present invention discloses a pharmaceutical composition comprising epinephrine and a penetration enhancer having the formula: CH3(CH2) n-1 [OCH2CH2] mOH, wherein n is an integer selected from 10-16, and m is an integer selected from 4-8.

[0011] In certain embodiments, n is equal to 12 and m is selected from 4, 7 and 8.

[0012] In certain embodiments, n is equal to 10 and m is equal to 6.

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

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

[0015] In the pharmaceutical composition according to any one of claims 1 to 5, the pH value of the pharmaceutical composition is lower 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 certain embodiments, the pharmaceutical composition is for nasal administration, or is used in a nasal administration device.

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

[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-tocopheryl polyethylene glycol succinate (vitamin E TPGS), butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA).

[0020] In certain embodiments, the concentration of epinephrine in the pharmaceutical composition ranges from 0.3% to 5% (w / v).

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

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

[0023] In certain embodiments, the effective permeability coefficient (Pe) of the pharmaceutical composition using the PAMPA test is greater than 3×10 - 6 cm / s.

[0024] In certain embodiments, the pharmaceutical compositions disclosed herein do not cause irreversible damage to the nasal mucosa.

[0025] In another aspect, the present invention discloses a method comprising administering an effective amount of a pharmaceutical composition to a subject.

[0026] In certain embodiments, the methods are used to treat an allergic reaction, particularly a Type I allergic reaction, in a subject.

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

[0028] In certain embodiments, the pharmaceutical composition is administered to the subject by nasal administration.

[0029] BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0032] Figure 3 Shown is the change in epinephrine concentration in plasma over time after administration of compositions having different epinephrine concentrations, and different types and concentrations of penetration enhancers.

[0033] Figure 4 The changes in the concentration of epinephrine in plasma over time after administration of compositions having different epinephrine concentrations and different types of penetration enhancers are shown.

[0034] Figure 5 The C of each subgroup in each dosing group is shown max (μU / mL).

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

[0036] Detailed description

[0037] definition

[0038] In the disclosure of the present invention, 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 more fully described with reference to the entire specification.

[0039] As used herein, unless otherwise specified, a quantifier preceding an element without expressing a plural number also includes a plural meaning.

[0040] As used herein, "and / or" refers to 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 ("or"). In addition, the present invention also contemplates that in certain disclosed embodiments, any feature or combination of features described herein may be excluded or omitted.

[0041] Unless otherwise stated herein, “comprises,” “comprising,” or similar terms are intended to represent a non-exclusive inclusion, such that the described list of elements or features includes not only those explicitly stated or listed, but may also include other elements or features not listed or explicitly stated.

[0042] It is to be understood that the present disclosure is not limited to the particular methodology, protocols, and reagents described, as such factors may vary depending upon the context in which they are used by one skilled in the art.

[0043] As used herein, "systemic administration" is defined as a mode of administration of a therapeutic product that is capable of producing extensive exposure of an active agent in an organism. As used herein, "topical administration" refers to a mode of administration that delivers an active agent directly to a target site in an organism. Topical administration does not exclude systemic pharmacological effects.

[0044] As used herein, the term "pharmaceutically acceptable" refers to materials such as carriers or diluents that do not abrogate the biological activity or properties of the therapeutic compound and are relatively nontoxic, i.e., the material can be administered to a subject without causing adverse biological effects or without adversely affecting any component of a composition containing the material. Pharmaceutically acceptable components include those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problems or complications within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0045] As used herein, "effective amount" refers to an amount of a pharmaceutical composition sufficient to significantly and positively alter the symptoms and / or conditions to be treated (e.g., to 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 employed, the specific pharmaceutically acceptable excipients and / or carriers used, and the knowledge and experience of the physician.

[0046] As used herein, "disease" or "disorder" refers to a condition for which treatment is needed and / or desired.

[0047] As used herein, the term "treatment" or "therapy" refers to improving a disease or condition, for example, slowing down, preventing or reducing the development of a disease or condition or reducing at least one clinical symptom thereof. For example, in certain embodiments, improving a disease or condition may include obtaining a beneficial or desired clinical outcome, including but not limited to one or more of: alleviating one or more symptoms, alleviating the extent of the disease, preventing or delaying the spread of the disease, preventing or delaying the recurrence of the disease, delaying or slowing the progression of the disease, improving the disease state, inhibiting the disease or the progression of the disease, inhibiting or slowing the disease or its progression, preventing its development, and alleviating (whether in part or in whole).

[0048] As used herein, the term "subject" refers to an animal. For example, in certain embodiments, the animal is a mammal. In certain embodiments, the animal includes humans, rodents, primates, felines, canines, equines, bovines, swine, sheep, goats, mammalian experimental animals, mammalian farm animals, mammalian sports animals, or mammalian pets. The animal can be male or female, and can be any appropriate age, including infants, young children, teenagers, adults, and old age. In certain embodiments, "individual" or "subject" refers to an animal that needs to treat a disease or illness. In certain embodiments, the animal to be treated can be referred to as a "patient", which means that the animal has been determined to suffer from a disease related to treatment or has enough risk to suffer from the disease. In certain 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 certain embodiments, the biological membrane is a nasal mucosa. Permeability can be measured by different permeability models, which can be performed in situ, in vivo or in vitro. Some exemplary permeability models will be discussed in the following sections.

[0050] As used herein, the term "permeation enhancer" refers to an excipient included in a formulation that is used to increase the permeability of an active pharmaceutical ingredient. Sometimes "permeation enhancers" are also referred to as "absorption enhancers" or "permeation enhancers". In certain embodiments, permeation enhancers promote nasal mucosal permeability. In certain embodiments, permeation enhancers promote the passage of paracellular pathways. In certain embodiments, permeation enhancers promote the passage of transcellular channels.

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

[0052] As used herein, the terms "allergic reaction," "anaphylactic reaction," "allergy," and "allergy" are used interchangeably.

[0053] As used herein, in the context of a pharmaceutical composition, the term "active ingredient" refers to any ingredient that provides pharmacological activity or other direct effects for diagnosis, cure, alleviation, treatment or prevention of disease, or affects the structure or any function of the body of the subject. For the purposes disclosed herein, the active ingredient of the adrenaline composition described herein is adrenaline.

[0054] As used herein, the term "therapeutic window" refers to the range of blood concentrations of a drug within which a desired effect occurs, below which there is little or no effect, and above which excessive toxicity occurs.

[0055] Overview

[0056] Epinephrine is the first-line treatment for type I allergic reactions. Type I allergic reactions, also known as anaphylaxis, involve immunoglobulin E (IgE)-mediated release of antibodies against soluble antigens, which leads to mast cell degranulation and the release of histamine and other inflammatory mediators. Type I allergic 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, 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 allergic reactions. At specific doses and routes of administration of epinephrine, alpha-adrenergic vasoconstriction reverses peripheral vasodilation, thereby relieving hypotension and reducing erythema, urticaria, and angioedema. The beta-adrenergic properties of epinephrine cause bronchodilation, increase myocardial output and contractility, and inhibit further release of mediators from mast cells and basophils.

[0057] When a patient experiences a Type I allergic reaction, epinephrine needs to be administered quickly. However, intramuscular epinephrine administration can be significantly delayed due to patient fear of needles, lack of training, and misunderstanding of the correct timing of administration. This delay can lead to adverse outcomes and may even result in the patient's death. In addition, intramuscular epinephrine may be mistakenly delivered into the vein or subcutaneously, resulting in serious side effects and a significant delay in the onset of action. Through nasal administration, these problems can be overcome, providing patients with a more convenient way to administer epinephrine in a timely and easy manner.

[0058] Nasal administration is a non-invasive route of administration in which a pharmaceutical composition is inhaled through the nasal cavity and absorbed through the nasal mucosa. The drug should first pass through the nasal mucosal layer and then through the epithelial layer to be absorbed to achieve a systemic effect. Drugs administered through the nasal cavity can be passively permeated through the paracellular pathway, or passively and actively permeated through the transcellular pathway. This process is largely affected by the lipophilicity of the compound. In addition to the passive transport pathway, carrier-mediated transport, transcellular transport through the cell membrane, and transport through tight junctions between cells are other possible pathways for drugs to penetrate the nasal mucosa (Arora et al., Permeability Issues in Nasal Drug Delivery, Drug Discovery Today Vol. 7-18, 2002). For polar and hydrophilic drugs, such as adrenaline, the main pathway is the paracellular pathway, which is related to the intercellular space and tight junctions.

[0059] The two main obstacles to drug absorption in nasal administration are low membrane permeability of polar drugs and rapid mucociliary clearance in the nasal cavity. Therefore, in order to achieve systemic absorption and pharmacokinetics comparable to intramuscular injection, it is crucial to find a more ideal permeation enhancer for intranasal formulations. The present invention provides such an effective permeation enhancer and its use.

[0060] Penetration enhancers

[0061] Polyoxyethylene alkyl ethers are nonionic surfactants consisting of a linear alkyl chain (containing n-1 methylene groups) and a hydrophilic part (containing m oxyethylene units). They have a general chemical formula of CH3(CH2) n-1 (OCH2CH2) m OH (Formula I). ​​They are also referred to 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 permeation enhancers in previous publications, polyoxyethylene alkyl ethers are a large family that includes an unlimited number of compounds, few of which are effectively and safely used as permeation enhancers in any commercially available nasal formulations. For example, polyoxyethylene-9-lauryl ether (C12E9) has been reported to cause severe multiple erosions of the nasal epithelium in dogs at a concentration of 1%, 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 unexpectedly found that a specific class of polyoxyethylene alkyl ethers exhibited significantly higher bioavailability, shorter onset of action, better safety and chemical stability in nasal administration of epinephrine relative to other formulations using C12E9 and other classes of permeation enhancers.

[0064] In one aspect, the present invention discloses a pharmaceutical composition comprising epinephrine and a penetration enhancer having the formula: CH3(CH2) n-1 [OCH2CH2] m OH, wherein 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 certain embodiments, n is 12 and m is selected from 4, 7 and 8.

[0066] In certain embodiments, n is 10 and m is 6.

[0067] For purposes of this disclosure, epinephrine includes the free form of epinephrine as well as pharmaceutically acceptable salts of epinephrine, including acid addition salts and base addition salts.

[0068] "Pharmaceutically acceptable acid addition salts" refer to salts which retain the biological effectiveness and properties of the free bases and do not possess biological or other undesirable properties, and which are formed by the free bases with inorganic or organic acids. Some exemplary acids include hydrochloric acid, tartaric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

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

[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 the compounds include isotopes of hydrogen, carbon, and oxygen, such as 2H, 3H, 11C, 13C, 14C, 15O, 17O, and 18O.

[0071] In certain embodiments, the concentration of the penetration enhancer ranges from 0.1% to 2.50% (v / v). In certain embodiments, the concentration of the penetration enhancer is 0.25% (v / v). In certain 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; grams of solute per 100 milliliters of solution (g / 100mL) have been used throughout this application. In this application, "v / v" means volume volume. Volume volume concentration is calculated by dividing the volume of the solute by the volume of the solution. For the purposes of the present invention, when 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 lower 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 and vice versa based on the density of the solute compound. The density of C10E6 is about 0.987 g / mL, the density of C12E4 is about 0.946 g / mL, the density of C12E7 is about 1.0 g / mL, the density of C12E8 is about 0.984 g / mL (measured at 35° C.), and the density of C12E9 is about 1.007 g / mL.

[0074] In certain embodiments, the pharmaceutical composition is used for nasal administration, or for a nasal administration device. In certain embodiments, nasal administration is performed by directly applying the pharmaceutical composition to the nasal mucosa. In certain 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 zone around the inferior turbinate (Grassin-Delyle et al., Pharmacology & Therapeutics 134: 366-379 (2012)). The nasal mucosa includes a layer of epithelial cells covering the nasal cavity. Drugs can be absorbed into the systemic circulation through the nasal mucosa.

[0076] Some exemplary nasal administration devices include steam inhalers, droppers, pipettes, squeeze bottles, spray pumps, atomizers, powder sprayers and insufflators (Djupesland, Drug Deliv. And Transl. Res. 3: 42-62, 2013). It is understood that those skilled in the art can select a suitable nasal administration device according to the dosage form, chemical properties, physical properties and other relevant considerations of a given pharmaceutical composition.

[0077] In certain embodiments, the concentration of epinephrine in the pharmaceutical compositions described herein ranges from 0.3% to 5% (w / v). In certain embodiments, the concentration of epinephrine in the pharmaceutical compositions 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 certain embodiments, the pharmaceutical composition described herein is in the form of a liquid. In certain embodiments, the pharmaceutical composition described herein is in the form of a spray. In certain 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 the present disclosure, the effective permeability coefficient (Pe) is used as an indicator of in vitro permeability testing in the parallel artificial membrane permeability assay (PAMPA). The PAMPA permeability test 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 oil layer in the middle maintains a strong and stable PAMPA membrane, and is ultra-thin to minimize the retention of the compound and the interference with the penetration of the compound. The test compound diluted in the buffer is placed at the supply end, and the compound enters the artificial membrane from the supply end and enters the receiving 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 protocol of PAMPA is described in Example 1.

[0081] The penetration enhancer disclosed in the present invention can effectively improve the permeability of the adrenaline composition. In certain embodiments, the absolute value of Pe of the pharmaceutical composition described in the present invention in the PAMPA test is higher than 3×10 -6 cm / s.

[0082] Furthermore, when the pharmaceutical composition is administered to a subject, the maximum blood concentration (C max ) and reach C max The time required (T max ) are two effective indicators of in vivo permeability. max The larger the value, the better the in vivo permeability. max The smaller it is, the better the in vivo permeability is.

[0083] Pharmacokinetic characteristics

[0084] Compared with previously disclosed penetration enhancers, namely C12E9, n-dodecyl-β-D-maltoside (DDM) and diethylene glycol monoethyl ether (DEGEE), the penetration enhancers disclosed in the present invention show more ideal pharmacokinetic characteristics. In certain embodiments, the pharmaceutical compositions disclosed in the present invention show better bioavailability. In certain embodiments, the pharmaceutical compositions disclosed in the present invention show faster absorption rate, i.e., shorter T maxIn certain embodiments, the pharmaceutical compositions disclosed herein show a greater early portion area under the curve (AUC).

[0085] The present disclosure also provides an unexpected discovery that the pharmaceutical compositions described in the present disclosure exhibit higher bioavailability compared to previously disclosed penetration enhancers (such as C12E9, DDM and DEGEE), especially at higher epinephrine doses that are closer to clinically used doses. Some pharmaceutical compositions described in the present disclosure even exhibit higher bioavailability. For example, compared to intramuscular administration of epinephrine, the exposure of the epinephrine composition containing C12E7 as a penetration enhancer is 2.7 times higher in nasal administration.

[0086] Compared with previously disclosed compositions (such as C12E9, DDM and DEGEE), the compositions disclosed in the present disclosure show a shorter T max and a larger early part of the area under the curve (AUC). Shorter T max and a larger early partial area under the curve AUC are more favorable characteristics because epinephrine is a life-saving drug used in emergency situations and a faster onset of epinephrine action is highly desirable (Assessment report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0087] Security

[0088] Compared with previously disclosed penetration enhancers (such as C12E9, DDM and DEGEE), the penetration enhancers disclosed in the present invention perform better in terms of safety. In certain embodiments, the pharmaceutical compositions disclosed in the present invention show better tolerability. In certain embodiments, the pharmaceutical compositions disclosed in the present invention do not cause irreversible damage to the nasal mucosa.

[0089] Epinephrine is a drug with a narrow therapeutic window. Doses exceeding the therapeutic window may lead to fatal side effects, such as cerebral hemorrhage, subarachnoid hemorrhage, etc. Therefore, an ideal penetration enhancer should be able to improve tolerance to epinephrine. In the disclosed pharmacokinetic study, all subjects in the C12E9 group who received nasal administration of an epinephrine composition containing C12E9 (1 mg / kg dose of epinephrine) died, with only 53.2% bioavailability compared to the intramuscular injection route. However, after nasal administration of an epinephrine composition containing C12E7 (1 mg / kg dose of epinephrine), only one subject was in a state of shock (recovered after cardiopulmonary resuscitation), and its bioavailability was more than 270% compared to the intramuscular injection route. In contrast, after nasal administration of 1 mg / kg doses of epinephrine compositions containing C12E23 and C16E10, respectively, the subjects' activities were significantly reduced, indicating that a certain degree of adverse reactions occurred. After nasal administration of an adrenaline composition containing C12E8, C12E4, or C10E6 (1 mg / kg dose of adrenaline), the subjects remained normal and active, and higher F values, shorter T max and a greater early partial drug-time curve area under the curve AUC. The above observations indicate that C12E7, C12E8, C12E4 and C10E6 are unexpectedly able to improve tolerance to adrenaline. When C12E7, C12E8, C12E4 and C10E6 are added to an adrenaline composition, the subject's tolerance to the composition can be improved. In certain embodiments, the adrenaline composition is administered to the subject via a nasal administration route.

[0090] For purposes of this disclosure, a subject having better "tolerance" to a drug or drug composition means that the subject has fewer adverse reactions, less severe adverse reactions, or is less prone to adverse reactions after administration of a composition containing the drug or drug composition (e.g., epinephrine or an epinephrine composition). Tolerance describes the ability of a subject to tolerate large doses of an active ingredient (e.g., epinephrine).

[0091] As used herein, irreversible damage refers to damage that cannot be self-recovered in a relatively short period of time. Due to the strong local pharmacological effects of adrenaline, when a penetration enhancer is used together with adrenaline, irreversible damage may become more serious. In the purpose disclosed in the present invention, the insulin absorption test is used to evaluate 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 basic conditions, the nasal epithelium severely limits the absorption of drugs with a molecular weight greater than 1kDa. Therefore, insulin with a molecular weight greater than 5kDa usually cannot pass through the nasal mucosa unless the nasal mucosa is damaged. After the penetration enhancer is exposed to the nasal cavity, insulin is administered nasally, and the C of insulin after nasal administration of insulin at different time points after nasal administration of a composition containing a drug (i.e., adrenaline) can be monitored. max The C and AUC of the composition were used to determine whether the composition caused any damage to the nasal mucosa of the subject and whether such damage, if any, was reversible. max and AUC indicated less damage to the nasal mucosa, whereas higher levels of insulin exposure resulted in a lower C max and AUC indicate that there is damage to the nasal mucosa. In the case of reversible nasal mucosal damage, the C max The AUC and C of insulin are high initially, then decline rapidly, returning to a significantly lower level within about 2 hours. max The AUC and AUC will remain at a high level for more than 2 hours. The detailed protocol of the insulin absorption test is shown in Example 5.

[0092] The present disclosure also provides an unexpected discovery that the pharmaceutical composition shows faster reversal of nasal mucosal damage under low pH (pH value below 7, for example, in the range of 4-6). When the pharmaceutical composition is formulated to have a pH value below 7, the pharmaceutical composition only causes reversible damage, which can be recovered faster. In contrast, many penetration enhancers known in the art and used in intranasal preparations, such as DDM or DEGEE, do not show the pH-sensitive trend of nasal mucosal damage disclosed in Example 5 in terms of nasal mucosal damage.

[0093] In addition, the pharmaceutical composition has better stability and permeability under low pH conditions, ie, a pH value lower than 7, such as in the range of 4-6.

[0094] Compared with previously disclosed compositions (e.g., compositions containing penetration enhancers such as C12E9, DDM, DEGEE), it was also unexpectedly found that the current compositions exhibited less nasal mucosal damage under low pH conditions, i.e., pH values ​​below 7, such as in the range of 4-6. For example, it has been reported that C12E9 causes severe nasal mucosal damage when adrenaline is administered via the nasal route in a composition having a pH of 7.4 (Bleske et al., Effect of Vehicle on the Nasal Absorption of Epinephrine during Cardiopulmonary Resuscitation, Pharmacotherapy, 16(6), 1039–1045 (1996)). However, the present disclosure provides a discovery that C12E9 causes much less nasal mucosal damage under low pH conditions, i.e., pH values ​​below 7, such as in the range of 4-6.

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

[0096] The present invention also provides a pharmaceutical composition comprising epinephrine and C12E9 as a penetration enhancer, wherein the pH value of the pharmaceutical composition is lower than 7, for example, ranging from 4 to 6.

[0097] stability

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

[0099] pH regulators, sometimes also called pH adjusters or acidity regulators, are substances used to adjust the pH of a pharmaceutical composition to a specific range. pH is an expression of the concentration of hydrogen ions in water. Specifically, pH is the concentration of hydrogen ions (H + ) Negative logarithm of concentration (mol / L): pH = -log 10 (H + ). In certain embodiments, the pH adjuster is an acid or a base. Some exemplary pH adjusters include hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, and ammonia.

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

[0101] Preservative is a substance added to a pharmaceutical composition to prevent adverse physical, chemical or biological changes. In certain embodiments, the preservative is a bactericide or antimicrobial. Some exemplary preservatives include benzalkonium chloride, chlorobutanol, butyl paraben, propyl paraben, benzethonium chloride, chlorocresol, phenol and benzoic acid.

[0102] Osmotic pressure regulators are usually small molecular weight water-soluble substances. Osmosis is the diffusion of water under osmotic pressure caused by molecular imbalance on both sides of a membrane. Osmotic pressure regulators can change the osmotic pressure across a cell membrane. Some example osmotic pressure regulators include sodium chloride, glucose, mannitol, sorbitol, lactose, phosphoric acid, and citric acid.

[0103] In certain embodiments, the pharmaceutical composition comprises an antioxidant. In certain embodiments, the antioxidant is selected from sodium bisulfite, sodium pyrosulfite (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 certain embodiments, the antioxidant is selected from sodium bisulfite and sodium pyrosulfite. In certain embodiments, the pharmaceutical composition comprises 0.2% sodium bisulfite. As shown in Example 4, when sodium bisulfite (as an antioxidant) is used together with EDTA-2Na (as a chelating agent), adrenaline preparations containing C10E6, C12E4, C12E7, C12E8 and C12E9 as penetration enhancers show significantly better color stability characteristics. The pharmaceutical composition containing a penetration enhancer and an antioxidant has better stability and permeability under low pH conditions, ie, a pH value lower than 7, such as in 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 maintain its chemical, physical, and biopharmaceutical properties over time. For the purposes of the present disclosure, stability is measured by the "content percentage" of the active ingredient in the pharmaceutical composition. As used herein, the content percentage 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, where the composition is stored under given environmental conditions (pH, temperature) starting from day 0 and throughout the accelerated stability test, where the content percentage is weight percentage.

[0105] In certain embodiments, the content percentage of the pharmaceutical composition under the conditions of pH 4.0 and 60° C. is not less than 90% on the 30th day.

[0106] method

[0107] In another aspect, the present invention discloses a method comprising administering an effective amount of the pharmaceutical composition to a subject. The administration route can be, for example, intravenous, intratumoral, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, and ocular.

[0108] In certain embodiments, the method is used to treat a subject suffering from type I allergic reactions. Type I allergic 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 allergic reactions include atopic diseases, i.e., excessive IgE-mediated immune responses (e.g., rhinitis, conjunctivitis, and dermatitis); and allergic diseases, i.e., immune responses to exogenous allergens (e.g., anaphylactic shock, urticaria, angioedema, food allergies, and drug allergies).

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

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

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

[0112] As used herein, AUC refers to the area under the blood concentration time curve, that is, the area under the curve of the blood concentration of the active ingredient or its metabolite over time after administration of a certain dose of the active ingredient (e.g., epinephrine). 0-∞ ” is the area under the concentration-time curve extrapolated to infinity after administration, “AUC 0-t ” is the area under the concentration-time curve from time zero to time t after drug administration, where t is the last time point at which concentration can be measured.

[0113] As used herein, bioavailability (F) describes the percentage of a drug's administered dose that reaches the systemic circulation. It has the meaning defined in 21 CFR § 320.1 (a). For the purposes of this disclosure, F specifically refers to the ratio of exposure per unit dose between the intranasal route of administration and the intramuscular route of administration. 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 herein, early fraction bioavailability (F 0-10 ) specifically refers to the ratio of AUC within the first 10 minutes after a unit dose is administered between the intranasal route and the intramuscular route. 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 description can be further described by the following non-limiting examples, in which standard techniques known to those skilled in the art and techniques similar to those described in these examples can be used where appropriate. It is understood that those skilled in the art will envision other embodiments consistent with the description provided by the present invention.

[0116] Example 1 Evaluation of PAMPA in vitro permeability

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

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

[0119] 200 μL of hydration solution (PION Inc., MA, USA) was added to each well of the supply port (P / N 110660, PION Inc., MA, USA). Then, the pre-coated PAMPA plate (P / N 120657, PION Inc., MA, USA) was immersed in the hydration solution overnight for full hydration.

[0120] The permeation enhancers (the concentrations listed in Table 2) were dissolved together with epinephrine (10 mg / mL, 50 mg / mL, 3 mg / mL) in a pH 4.0 aqueous buffer to prepare the designed compositions. The pH 4.0 epinephrine aqueous solution (without permeation enhancer) was used as the experimental control group. 200 μL of each composition solution was accurately added to each well of the supply end, and 200 μL of PRISMA TM Buffer was added to each well of the receiving end. The PAMPA artificial membrane was then placed between the two plates of the supply and receiving ends to start the in vitro permeability evaluation experiment. The permeation experiment of each composition was repeated in parallel in 4 wells. The PAMPA kit including the supply end, PAMPA artificial membrane and the receiving end was covered and incubated at 37°C for 5 hours. Antioxidants were also used to prevent oxidation reactions during the experiment. After incubation, the concentration of epinephrine in the supply end and receiving end solutions was determined separately using UPLC using the method described in Table 1.

[0121] Table 1. UPLC analytical method for in vitro permeability assessment of epinephrine using the PAMPA assay

[0122]

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

[0124] Table 2. In vitro permeability test results of epinephrine solutions containing different types of permeation enhancers

[0125]

[0126]

[0127] Example 2 Pharmacokinetic (PK) study of compositions containing penetration enhancers after nasal administration The preparation method of different formulation compositions for intranasal administration is as follows: first, an aqueous solution containing 0.25% penetration enhancer, 0.2% sodium bisulfite and 0.9% sodium chloride is prepared, and then epinephrine is dissolved to reach a concentration of 10 mg / mL or 3 mg / mL, and then all formulation solutions are adjusted to pH 4 and refrigerated. The liquid composition containing epinephrine and penetration enhancer is administered nasally according to the method of Table 3.

[0128] The prepared solution composition was administered intranasally to Sprague-Dawley (SD) rats, and then blood was sampled into 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90 and 120 minutes after the 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 fully vortexed and stored in an ice water bath to await pretreatment. If the sample was not pretreated and analyzed on the day of the PK experiment, the sample was stored in a -80°C refrigerator. For frozen samples, equilibrate 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, 180 μL of internal standard solution (preparation method see Table 5) was added, and the mixed sample was vortexed for 5 minutes and centrifuged for 5 minutes at 10,000 rpm and 4°C. 130 μL of supernatant was taken into a 96-well plate, 130 μL of ultrapure water was added, and then vortexed for 5 minutes and centrifuged for 5 minutes at 4,000 rpm and 4°C. 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 of rat PK study of different formulation compositions

[0130]

[0131] Table 4. Epinephrine HPLC analysis method

[0132]

[0133]

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

[0135]

[0136] Table 6. Adrenal mass spectrometry analysis methods

[0137]

[0138] Table 7. Adrenal mass spectrometry analysis methods

[0139]

[0140] The changes in plasma adrenaline concentration over time in the 10 mg / mL adrenaline concentration prescription (dosage 1 mg / kg) group are shown in Table 8 and Figure 1As shown. In the intramuscular injection PK study, 3 SD rats were used in each group, and the adrenaline concentration was 1 mg / mL and the dose was 0.1 mg / kg. For the nasal administration PK study, 2 SD rats were used in each group, and the adrenaline concentration was 10 mg / mL and the dose was 1 mg / kg. In Examples 2 and 3, "NA" means that adrenaline was not detected in the plasma at a specific time point or the parameter could not be calculated because adrenaline was not detected at a specific time point. MaS Studio (v1.5.3.10) was used to calculate the PK parameters, i.e., T max , C max , AUC, bioavailability (F 0-t ) and the early fraction bioavailability (F 0-10 ), as shown in Table 9. It was observed that when the polyoxyethylene alkyl ether has 9-15 methylene groups and 4-10 oxyethylene units, C max , AUC and bioavailability were significantly higher.

[0141] Table 8. Changes of plasma adrenaline concentration over time after nasal 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 (the analysis method in Table 5 was used for samples with *, and the analysis method in Table 5 was used for samples with The samples were analyzed using the analysis method in Table 6)

[0146]

[0147] The changes in plasma adrenaline concentration over time in the 3 mg / mL adrenaline concentration prescription group (much lower than the 0.3 mg / kg dose used clinically) are shown in Tables 10 and Figure 2 MaS Studio (v1.5.3.10) was used to calculate the PK parameters, namely T max , C max , AUC and bioavailability are shown in Table 11. It was observed that when the polyoxyethylene alkyl ether has 9-15 methylene units 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 More ideal because for treating type I anaphylaxis, a faster onset of epinephrine is required.

[0148] Table 10. Changes of epinephrine concentration in plasma 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 (samples with * were used

[0151] Table 5 Analysis method, The samples were analyzed using the analysis method in Table 6)

[0152]

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

[0154] The preparation process of the composition for intranasal administration 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, and 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 nasally according to Table 12.

[0155] The compositions of different prescriptions were dripped into the nasal cavity of SD rats, and blood was sampled into 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90 and 120 minutes after the administration was completed. Prepare a 10% (w / v) sodium metabisulfite aqueous solution and add it to the sample at a volume ratio of 9:1 to prevent oxidation. Vortex the sample thoroughly and store it in an ice water bath to wait for pretreatment. If the sample pretreatment and analysis are not performed on the day of the PK experiment, store the sample in a -80°C refrigerator. For frozen samples, equilibrate at room temperature before further pretreatment. After the sample thaws, take 20 μL of plasma sample and transfer it to a 1.5 mL polyethylene centrifuge tube, add 180 μL of internal standard solution (preparation method see Table 5), vortex the mixed sample for 5 minutes and centrifuge it for 5 minutes at 10,000 rpm and 4°C. 130 μL of supernatant was taken into a 96-well plate, 130 μL of ultrapure water was added, and then vortexed for 5 minutes and centrifuged for 5 minutes at 4,000 rpm and 4°C. 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] The concentration of adrenaline in plasma at different time points after administration of the composition with different adrenaline concentrations, penetration enhancer types and concentrations is shown in Table 13 and Figure 3 It was observed that the tested polyoxyethylene alkyl ethers had obvious permeation enhancement ability in the concentration range of 0.1 mg / mL to 2.5 mg / mL. The PK parameters, namely T max , C max , AUC, F 0-t and F 0-10 , see Table 14. It was observed that, when the administration volume was constant, with the increase of the penetration enhancer concentration and adrenaline concentration, C max Compared with the groups containing DDM and DEGEE, polyoxyethylene alkyl ethers showed higher C max , AUC and bioavailability, suggesting that it has better penetration enhancement ability. A shorter T max , indicating that adrenaline takes effect more quickly.

[0159] Table 13. Changes in plasma epinephrine concentration over time after administration of compositions with different epinephrine concentrations, types and concentrations of penetration 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 penetration enhancers after nasal administration The preparation method of different formulation compositions for intranasal administration is as follows: first, an aqueous solution containing 0.25% penetration enhancer, 0.2% sodium metabisulfite and 0.9% sodium chloride is prepared, and then epinephrine is dissolved to achieve an epinephrine concentration of 2.5 mg / mL, 10 mg / mL or 20 mg / mL, and then all formulation solutions are adjusted to pH 4 and refrigerated. The liquid composition containing epinephrine and penetration enhancer is administered to the nasal cavity of beagle dogs according to Table 15.

[0166] The composition was administered intranasally to beagles, and blood was collected at 1, 5, 10, 15, 20, 30, 60, 90 and 120 minutes after the administration. Whole blood samples were collected in anticoagulant tubes containing EDTA-K2 and centrifuged at 1524g 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. Take 100 μL of sample, add 20 μL of water (containing 0.1% acetic acid, 50 ng / mL epinephrine-D6) as a protein precipitant, then add 250 μL of PBA, vortex and mix well, add 400 μL of TOAB, vortex the mixture for 10 minutes, and then centrifuge at 13000 rpm for 5 minutes. Take 300 μL of the supernatant in 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. Take the lower layer solution, mix 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 Table 16 and Table 7, respectively.

[0167] Table 15. Experimental design of beagle dog PK study with different formulation compositions

[0168]

[0169]

[0170] Table 16. Epinephrine HPLC analysis method

[0171]

[0172] Table 17 and Figure 4 The figure shows the changes of plasma epinephrine concentration over time at different time points after administration in the prescription groups with different dosage concentrations and types of penetration enhancers. The corresponding pharmacokinetic parameters (i.e., T max , C max , AUC, bioavailability relative to intramuscular injection route F 0-t and the early fraction bioavailability F within the first 10 minutes after administration 0-10 ) was calculated using Phoenix WinNonlin 8.3 software, see Table 18. Compared to commercial preparations with DDM as a penetration enhancer, the C12E7 group still showed higher bioavailability even at lower epinephrine concentrations and dosages. The C12E7 group also showed an early portion AUC that was nearly 3 times higher than the DDM group (the currently used commercial preparation). It is worth noting that when epinephrine is used in emergency situations to treat type I allergic reactions (including anaphylactic shock), a higher early portion AUC is very desirable.

[0173] Table 17. Changes in plasma epinephrine concentration over time after nasal 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 Evaluation of composition stability

[0178] Epinephrine was dissolved in an aqueous solution containing 0.25% of different penetration enhancers and 0.9% sodium chloride to prepare an intranasal composition with an epinephrine concentration of 10 mg / mL. To prepare a formulation containing an antioxidant and a preservative, epinephrine was dissolved in an aqueous solution containing 0.1% to 1.0% of different penetration enhancers, 0.2% antioxidant, 0.1% preservative and 0.9% sodium chloride. To prepare a formulation containing an antioxidant under different pH conditions, epinephrine was dissolved in an aqueous solution containing 1.0% of different penetration enhancers, 0.2% antioxidant and 0.9% sodium chloride, and the pH was adjusted to 4.0, 5.0, 6.0 and 7.0, respectively. For the composition containing 1.0% C12E9, a sample with a pH of 7.4 was also prepared, which contained 0.2% antioxidant and phosphate buffered saline (PBS). The stability test design of the composition is shown in Table 19.

[0179] Table 19. Formulation stability test design

[0180]

[0181] The liquid composition was dispensed into vials, which were then capped and placed in an oven for accelerated stability testing. <391> EPINEPHRINE ASSAY), the epinephrine content in different compositions was quantified by high performance liquid chromatography (HPLC) before and after the accelerated stability test, and the analysis method is shown in Table 20. The preparation method of phosphate buffer (pH 2.8) is as follows: 5.0 g / L of potassium dihydrogen phosphate and 2.6 g / L of sodium octane sulfonate are added to water, and then the pH is adjusted to 2.8. The phosphate buffer is filtered through a 0.45 μm filter before use.

[0182] Table 20. Epinephrine HPLC analysis method

[0183]

[0184]

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

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

[0187]

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

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

[0190]

[0191] The results of the accelerated stability tests of the compositions containing 10 mg / mL epinephrine, 0.1% or 1.0% of different penetration enhancers, 0.2% of different antioxidants, and 0.1% of different preservatives at 60°C are shown in Tables 23 and 24. When two preservatives were added at the same time, no significant stability difference was found, indicating good chemical compatibility.

[0192] Table 23. Changes in adrenaline content (%) of 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] The results of stability tests of compositions 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 acceleration at 60°C are shown in Table 25. It was observed that the antioxidant sodium bisulfite significantly improved the stability of the composition in the pH range of 4.0 to 6.0, wherein the content of epinephrine was still above 95% after 7 days of acceleration at 60°C. However, when the pH value was equal to or higher than 7.0, the stability of the composition decreased rapidly. In addition, it was observed that sodium bisulfite only improved the stability of the composition in the pH range of 4.0 and pH 6.0; when the pH value 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 (%) of different compositions after acceleration at 60°C

[0198]

[0199]

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

[0201] Since penetration enhancers and epinephrine are likely to cause nasal mucosal damage, experiments are designed and conducted to evaluate the nasal mucosal damage after nasal administration of compositions containing epinephrine and different penetration enhancers. Unless the nasal mucosa is damaged, human insulin can hardly be absorbed by the nasal mucosa. 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), after nasal administration of a composition containing a penetration enhancer and epinephrine, the absorption of insulin by the nasal mucosa at different times is used to assess nasal mucosal damage. Absorption of more insulin by the nasal cavity indicates that the nasal mucosa is damaged, while the absence of insulin absorption indicates that the nasal mucosa is intact.

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

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

[0204]

[0205]

[0206]

[0207]

[0208] Unexpectedly, the pharmacokinetic parameters of insulin show that the composition containing C12E9 exhibits irreversible damage at higher pH values ​​(e.g., pH 7.4), while when the pH of the composition is lower (e.g., pH 4.0), milder and reversible damage is observed. Compared with the irreversible nasal mucosal damage observed when using DDM, DEGEE and C12E9 (high pH), all compositions containing polyoxyethylene alkyl ethers disclosed in this application show that the nasal mucosa recovers to a normal state at a faster rate (within 2 hours) at a lower pH. In addition, it is also observed that the compositions containing polyoxyethylene alkyl ethers tested have higher insulin AUC and C at 0 hours. max , indicating that the drug absorption is faster than DDM, DEGEE and C12E9 (high pH). Therefore, especially when used in an appropriate pH environment, the polyoxyethylene alkyl ether disclosed in this application can be used as a safer (such as faster recovery of nasal mucosa) and more effective (such as faster absorption of drugs after administration) penetration enhancer compared with 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 epinephrine and a penetration enhancer having the formula: CH3(CH2) n-1 [OCH2CH2] m OH, wherein 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, 8 and 9.

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 4, wherein the concentration of the penetration enhancer is 0.25% (v / v).

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pH value of the pharmaceutical composition is below 7.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the pH value of the pharmaceutical composition ranges from 4 to 6.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is for nasal administration, or for a nasal administration device.

9. The pharmaceutical composition according to any one of claims 1 to 8, further comprising at least one pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is a pH regulator, an antioxidant, a preservative or an osmotic pressure regulator.

10. The pharmaceutical composition according to claim 9, wherein the antioxidant is selected from sodium bisulfite, sodium metabisulfite (SMB), propyl gallate (PG), sodium sulfite, ascorbic acid (VC), methionine, α-lipoic acid, cysteine ​​(CYS), D-α-tocopheryl polyethylene glycol succinate (vitamin E TPGS), butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA).

11. The pharmaceutical composition according to claim 9 or 10, wherein the antioxidant is selected from sodium bisulfite and sodium metabisulfite.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the concentration of adrenaline in the pharmaceutical composition ranges from 0.3% to 5% (w / v).

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the dosage form of the pharmaceutical composition is a liquid or a spray.

14. The pharmaceutical composition according to any one of claims 1 to 13, wherein the recovery rate of the pharmaceutical composition at pH 4.0 and 60°C on the 30th day is not less than 90% by weight.

15. The pharmaceutical composition according to any one of claims 1 to 14, wherein the absolute value of Pe of the pharmaceutical composition in the PAMPA test is higher than 3×10 -6 cm / s.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the pharmaceutical composition does not cause irreversible damage to the nasal mucosa.

17. A method comprising administering to a subject an effective amount of the pharmaceutical composition according to any one of claims 1 to 16.

18. The method of claim 17, wherein the method is used to treat a subject suffering from a type I hypersensitivity reaction.

19. The method of claim 17, wherein the method is used to treat a condition selected from the group consisting of allergic asthma, allergic conjunctivitis, allergic rhinitis, anaphylactic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

20. The method according to any one of claims 17 to 19, wherein the pharmaceutical composition is administered to the subject by nasal administration.

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