Novel formulations of epinephrine and their uses

Abstract

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

Description

[Technical Field]

[0001] The present invention relates to novel formulations of epinephrine containing polyoxyethylene alkyl ethers as penetration enhancers, methods of administering the formulations, and uses thereof.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of International Patent Application PCT / CN2022 / 126776, filed October 21, 2022, the entire contents of which are incorporated herein by reference. [Background technology]

[0003] Epinephrine is a hormone and neurotransmitter secreted by the adrenal glands. Epinephrine can also be chemically synthesized. Epinephrine is used as a drug to treat a variety of conditions.

[0004] Specifically, epinephrine is the first-line drug for treating type I allergic reactions, including allergic shock. It has been estimated that up to 2% of the world's population will experience an allergic reaction at some point in their lives, with the trend toward an upward trend (Simons et al., The World Allergy Organization Journal, 4(2):13-37(2006)). Allergic shock is a severe, potentially fatal allergic reaction and medical emergency that develops quickly and requires immediate medical attention, regardless of whether emergency medications are used at the scene (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, epinephrine auto-injectors have been developed to allow for faster self-administration of epinephrine via intramuscular injection in emergency situations (Mylan Specialty LP, "EPIPEN® Epinephrine Injection, EPIPEN Jr® Epinephrine Injection" (FDA product label (archived February 1, 2014 (PDF) retrieved January 22, 2014)). However, epinephrine auto-injectors have a relatively high rate of risk, including misuse as a subcutaneous or intravenous injection or administration of an incorrect dose (Bilo, M. Beatrice., Anaphylaxis caused by Hymenoptera stings: from epidemiology to treatment, Allergy 66, pages 35-37 (2011)). Epinephrine injectors are structurally complex and prone to frequent mechanical failure, making this life-saving medication less reliable than expected.Furthermore, patients (whether they have needle phobia or not) are reluctant to use auto-injectors in public places and invariably delay using them until symptoms become severe, resulting in delayed treatment and ultimately ineffective reversal of the rapid progression of Type I allergic reactions.

[0005] Given the challenges facing epinephrine auto-injectors, a more reliable epinephrine delivery method with higher patient compliance is needed. Nasal administration is a promising route of administration, and numerous epinephrine nasal spray formulations have been developed. However, conventional epinephrine nasal spray formulations suffer from poor absorption, nasal mucosal damage, and poor chemical stability. For example, compared with intramuscular injection, conventional epinephrine nasal spray formulations still require more than three times the dose (0.3 mg vs. 1 mg) even with the use of penetration enhancers (Australian Patent No. AU2019217643B2). It is well known that epinephrine has a very narrow therapeutic window and poor absorption, which is always closely related to differences in absorption rates. Therefore, if relatively high doses are used to address the problem of poor absorption, it can lead to drug overdose in patients with relatively high absorption rates, potentially causing serious adverse reactions such as cerebral hemorrhage, hemiplegia, subarachnoid hemorrhage, and respiratory distress. A similar situation has been identified as a major safety risk of another drug that needs to be further improved (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 stated that the early portion AUC value, especially the AUC in the first 10 minutes after administration, is very important for the therapeutic effect of epinephrine nasal sprays, and that T max There is also a correlation between T and T at the same epinephrine dose. maxshould be equivalent to intramuscular (IM) or subcutaneous (SC) injections, and C max The Committee for Medicinal Products for Human Use (Committee for Human Use) clearly indicated that the AUC and total AUC are considered to be the parameters most correlated with the safety and efficacy of a drug. Neffy is an epinephrine nasal spray, and a new drug application (NDA) was submitted to the EMA, but it was not approved due to a relatively small early partial AUC compared to intramuscular injection (IM) (Assessment Report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0006] Furthermore, the simultaneous use of a penetration enhancer and epinephrine in a nasal spray formulation has been reported to 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)). Furthermore, epinephrine is highly susceptible to oxidation, making it extremely difficult to maintain the stability of epinephrine formulations (GB West, Oxidation of Adrenaline in Alkaline Solution, British Journal of Pharmacology and Chemotherapy, Volume 2, Issue 2, pp. 121-130, 1947). Therefore, the development of an epinephrine nasal spray formulation with higher bioavailability, less nasal mucosal damage, less individual variability, faster absorption, and better chemical stability is essential. Summary of the Invention

[0007] The present invention discloses a novel epinephrine formulation containing a polyoxyethylene alkyl ether as a penetration enhancer, a method for administering the epinephrine formulation, and its use. The formulation disclosed in the present invention can have higher bioavailability, a faster absorption rate, better chemical stability, and less damage to the nasal mucosa.

[0008] According to one aspect, the present invention discloses a pharmaceutical composition, the composition comprising epinephrine and a compound of formula CH3(CH2) n-1 [OCH2CH2] m and a penetration enhancer having an OH group, wherein n is an integer selected from 10 to 16, and m is an integer selected from 4 to 8.

[0009] In one embodiment, n is equal to 12 and m is selected from 4, 7, and 8.

[0010] In one embodiment, n is equal to 10 and m is equal to 6.

[0011] In certain embodiments, the concentration range of the penetration enhancer in the pharmaceutical compositions described herein is 0.1% to 2.50% (v / v).

[0012] In certain embodiments, the concentration of the penetration enhancer in the pharmaceutical compositions described herein is 0.25% (v / v).

[0013] In the pharmaceutical composition according to any one of claims 1 to 5, the pH value of the pharmaceutical composition is less than 7.

[0014] In the pharmaceutical composition according to any one of claims 1 to 6, the pH range of the pharmaceutical composition is 4-6.

[0015] In certain embodiments, the pharmaceutical composition is for use in nasal administration or in a nasal administration device.

[0016] In certain embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable auxiliary agent, wherein the pharmaceutically acceptable auxiliary agent is a pH adjusting agent, an antioxidant, a preservative, or an osmolality adjusting agent.

[0017] 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-alpha-tocopheryl polyethylene glycol succinate (vitamin E TPGS), butylhydroxytoluene (BHT), and butylhydroxyanisole (BHA).

[0018] In one embodiment, the concentration of epinephrine in the pharmaceutical composition is in the range of 0.3% to 5% (w / v).

[0019] In certain embodiments, the pharmaceutical composition is in the form of a liquid or spray.

[0020] In one embodiment, the pharmaceutical composition has a recovery rate of 90% by weight or more after 30 days under conditions of pH 4.0 and 60°C.

[0021] In one embodiment, the effective permeability coefficient (Pe) of the pharmaceutical composition as measured using PAMPA is 3×10 -6 More than cm / s.

[0022] In certain embodiments, the pharmaceutical compositions disclosed in the present invention do not cause irreversible damage to the nasal mucosa.

[0023] According to another aspect, the present invention discloses a method comprising administering to a subject an effective amount of the pharmaceutical composition.

[0024] In one embodiment, the method is used to treat an allergic reaction, particularly a type I allergic reaction, in a subject.

[0025] In certain embodiments, the allergic reaction is selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, allergic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

[0026] In certain embodiments, the pharmaceutical composition is administered to the subject by intranasal administration. [Brief explanation of the drawings]

[0027] [Figure 1] The graph shows the change in plasma epinephrine concentration over time in the epinephrine concentration 10 mg / mL (dose 1 mg / kg) group. [Figure 2] The graph shows the change over time in plasma epinephrine concentration in the group with an epinephrine concentration of 3 mg / mL (the epinephrine concentration of 3 mg / mL is far lower than the clinically used concentration, and the dose was 0.3 mg / kg). [Figure 3] 1 shows the change in plasma epinephrine concentration over time after administration of compositions having different epinephrine concentrations and different penetration enhancer types and concentrations. [Figure 4] 1 shows the change in plasma epinephrine concentration over time after administration of compositions having different epinephrine concentrations and different types of penetration enhancers. [Figure 5] Cmax (μU / mL) for each subgroup in each administration group is shown. [Figure 6] The AUC (min×μU / mL) for each subgroup in each administration group is shown. DETAILED DESCRIPTION OF THE INVENTION

[0028] definition In disclosing the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those of ordinary skill in the art. Correspondingly, terms defined herein are more fully explained by reference to the specification as a whole.

[0029] As used herein, unless otherwise indicated, quantitative words without a plural designation before the element also include a plural meaning.

[0030] As used herein, "and / or" refers to or includes any and all possible combinations of one or more of the listed items, as well as no combinations when understood as alternatives ("or"). Furthermore, it is contemplated that the invention may, in certain disclosed embodiments, exclude or omit any feature or combination of features described herein.

[0031] Unless otherwise expressly stated in the specification, the terms "comprises," "contains," or similar terms are intended to mean a non-exclusive inclusion, so that a described list of elements or features not only includes those expressly indicated or listed, but may also include other elements or features not listed or expressly indicated.

[0032] It is to be understood that the present disclosure is not limited to the particular methodology, protocols, and reagents described, which may vary depending on the circumstances in which one of skill in the art uses them.

[0033] As used herein, "systemic administration" is defined as a method of administration of a therapeutic product that is widely exposed and capable of producing an active agent in vivo. As used herein, "local administration" refers to an administration method that delivers an active agent directly to a target site in the body. Local administration does not exclude systemic pharmacological effects.

[0034] As used herein, the term "pharmaceutically acceptable" refers to a material, such as a carrier or diluent, that does not abolish the biological activity or properties of the therapeutic compound and is relatively non-toxic, i.e., the material can be administered to a subject without causing adverse biological effects or adversely affecting any components in the composition containing it. Pharmaceutically acceptable ingredients include compounds, materials, compositions and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, and that are consistent with a reasonable benefit / hazard ratio.

[0035] As used herein, "effective amount" refers to that amount of a pharmaceutical composition sufficient to significantly and positively alter the symptoms and / or condition being treated (e.g., provide a positive clinical response). The effective amount of a pharmaceutical composition will vary depending on factors such as the particular condition being treated, the severity of the condition, the duration of treatment, the nature of any concurrent treatments, the specific composition used, the particular pharmaceutically acceptable adjuvants and / or carriers used, and the knowledge and experience of the physician.

[0036] As used herein, a "disease" or "disorder" refers to a condition for which treatment is necessary and / or desired.

[0037] As used herein, the term "treatment" or "treatment" refers to ameliorating a disease or condition, e.g., alleviating, preventing, or reducing the progression of a disease or condition, or reducing at least one clinical symptom thereof. For example, in certain embodiments, ameliorating a disease or condition can include obtaining a beneficial or desired clinical outcome, including, but not limited to, one or more of: ameliorating one or more symptoms, reducing the extent of the disease, preventing or slowing the spread of the disease, preventing or slowing the recurrence of the disease, slowing or alleviating the progression of the disease, improving the disease state, inhibiting the disease or disease progression, inhibiting or alleviating the disease or its progression, preventing its progression, and remission (whether partial or total).

[0038] 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 a human, rodent, primate, feline, canine, equine, bovine, porcine, sheep, caprine, mammalian laboratory animal, mammalian livestock animal, mammalian sport animal, or mammalian pet. The animal may be male or female and of any appropriate age, including infant, toddler, adolescent, adult, and elderly. In some embodiments, an "individual" or "subject" refers to an animal in need of treatment for a disease or condition. In some embodiments, an animal receiving treatment may be referred to as a "patient," meaning that the animal has been determined to suffer from or is at substantial risk of suffering from the disease relevant to the treatment. In some embodiments, such an animal is a human, such as a human patient.

[0039] As used herein, "penetration" refers to the ability of a pharmaceutical composition to penetrate a biofilm. In one embodiment, the biofilm is the nasal mucosa. Penetration can be measured by various permeability models that can be performed in situ, in vivo, or in vitro. Some exemplary permeability models are discussed below.

[0040] As used herein, the term "penetration enhancer" refers to an adjuvant included in a formulation to increase the permeability of an active drug ingredient. A "penetration enhancer" may also be referred to as an "absorption enhancer" or "permeation enhancer." In certain embodiments, a penetration enhancer enhances nasal mucosal permeability. In certain embodiments, a penetration enhancer enhances passage through cell bypasses. In certain embodiments, a penetration enhancer enhances passage through transcellular channels.

[0041] As used herein, the terms "nasal administration" or "intranasal administration" refer 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.

[0042] As used herein, "allergic reaction," "allergic reaction," "allergy," and "hypersensitivity" are used interchangeably.

[0043] As used herein, in the context of pharmaceutical compositions, the term "active ingredient" refers to any ingredient that provides pharmacological activity or other direct action to diagnose, cure, ameliorate, treat, or prevent disease, or to affect the body structure or any function of a subject. For purposes disclosed herein, the active ingredient of the epinephrine compositions described herein is epinephrine.

[0044] As used herein, the term "therapeutic window" refers to the range of blood drug concentrations of a drug above which a desired effect occurs, below which there is little effect, and above which there is excessive toxicity.

[0045] overview Epinephrine is the first-line drug for treating type I allergic reactions. Type I allergic reactions, also known as immediate-type allergic reactions, involve immunoglobulin E (IgE)-mediated antibody release 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, 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., allergic shock, urticaria, angioedema, and food and drug allergies). By binding to multiple cellular receptors, epinephrine helps increase blood flow, relax lung muscles, and inhibit the release of chemicals that cause allergic reactions. At certain doses and administration routes, the vasoconstrictor effect of α-epinephrine reverses peripheral vasodilation, thereby reducing hypotension and alleviating erythema, urticaria, and angioedema. Epinephrine's β-epinephrine properties result in bronchodilation, increase myocardial emptying and contractility, and inhibit further release of mediators by mast cells and basophils.

[0046] When a patient has a type I allergic reaction, rapid administration of epinephrine is necessary. However, intramuscular injection of epinephrine can be significantly delayed due to the patient's fear of needles, insufficient training, and misunderstanding of the correct administration timing. This delay can lead to poor outcomes and even death. Furthermore, intramuscular injection of epinephrine can be mistakenly delivered intravenously or subcutaneously, resulting in severe side effects and significantly delayed onset of efficacy. Nasal administration overcomes these problems and provides a more convenient method for patients, allowing epinephrine to be administered immediately and easily.

[0047] Nasal administration is a non-invasive route of administration in which pharmaceutical compositions are inhaled through the nasal cavity and absorbed through the nasal mucosa. To achieve systemic effects, drugs must first pass through the nasal mucosa layer and then through the epithelial layer for absorption. Nasally administered drugs may passively permeate through a cell bypass pathway or passively and actively permeate through a transcellular pathway. This process is greatly influenced by the lipophilicity of the compound. In addition to passive transport, vector-mediated transport, transcellular transport, and transport via tight intercellular junctions are all other possible routes for drugs to pass through 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 epinephrine, the primary route is the cell bypass channel associated with the intercellular gap and tight junctions.

[0048] Two major obstacles to drug absorption in nasal administration are the low membrane permeability of polar drugs and the rapid clearance rate of nasal mucosal hairs. Therefore, it is important to find a more ideal penetration enhancer for intranasal formulations to achieve systemic absorption and pharmacokinetics comparable to those of intramuscular injection. The present invention provides such an effective penetration enhancer and its use.

[0049] penetration enhancers Polyoxyethylene alkyl ethers are nonionic surfactants consisting of a linear alkyl chain (containing n-1 methylene groups) and a hydrophilic portion (containing m oxyethylene units). They have the general formula CH3(CH2) n-1 (OCH2CH2) m OH (Formula I). ​​They are also called CnEm, where n represents the number of carbons in the alkyl chain and m represents the number of ethylene oxide units in the hydrophilic portion.

[0050] [ka]

[0051] Polyoxyethylene alkyl ethers have been reported as penetration enhancers in previous publications, but this is a large family of compounds, few of which have been effectively and safely used 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 in the nasal epithelium of dogs, suggesting that polyoxyethylene alkyl ethers may not be suitable for nasal epithelial 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 surprisingly found that certain types of polyoxyethylene alkyl ethers exhibit significantly higher bioavailability, shorter onset times, and better safety and chemical stability in nasal epinephrine administration compared to other formulations using C12E9 and other types of penetration enhancers.

[0052] According to one aspect, the present disclosure provides a compound comprising epinephrine and a compound of formula CH3(CH2) n-1 [OCH2CH2] mand a penetration enhancer having an 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.

[0053] In one embodiment, n is 12 and m is selected from 4, 7, and 8.

[0054] In one embodiment, n is 10 and m is 6.

[0055] For purposes disclosed in this invention, epinephrine includes the free form of epinephrine and pharmaceutically acceptable salts of epinephrine, including acid addition salts and base addition salts.

[0056] "Pharmaceutically acceptable acid addition salt" refers to a salt formed from a free base with an inorganic or organic acid that retains the biological effectiveness and properties of the free base, without possessing biologically or other undesirable properties. Some exemplary acids include hydrochloric acid, tartaric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

[0057] "Pharmaceutically acceptable base addition salts" refer to salts that retain the biological effectiveness and properties of the free acids without possessing biologically or other undesirable 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.

[0058] The disclosure of the invention 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 a different atomic mass or mass number. 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.

[0059] 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).

[0060] As used herein, "w / v" refers to 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, and in this application, grams of solute per 100 milliliters of solution (g / 100 mL) are always used. In this application, "v / v" refers to volumetric volume. Volumetric volume concentration is calculated by dividing the volume of the solute by the volume of the solution. For purposes of the present invention, when a concentration is indicated as "w / v or v / v," it means that for a solid solute, the concentration is w / v, and for a liquid solute, the concentration is v / v. In the case of a compound that is solid at room temperature but has a relatively low melting point, such as C12E8, the v / v concentration is obtained by first melting the compound and then preparing a solution.

[0061] It should be understood that one skilled in the art can convert w / v to v / v, and vice versa, depending on the density of the solute compound: C10E6 has a density of about 0.987 g / mL, C12E4 has a density of about 0.946 g / mL, C12E7 has a density of about 1.0 g / mL, C12E8 has a density of about 0.984 g / mL (measured at 35° C.), and C12E9 has a density of about 1.007 g / mL.

[0062] In some embodiments, the pharmaceutical composition is administered nasally or in a nasal administration device. In some embodiments, nasal administration is achieved by applying the pharmaceutical composition directly to the nasal mucosa. In some embodiments, nasal administration is achieved by a subject inhaling the pharmaceutical composition into the nasal cavity.

[0063] The nasal cavity is divided into the vestibule, antral cavity, inferior turbinate, middle turbinate, and superior turbinate. After nasal administration, drug deposition occurs primarily in the respiratory area around the inferior turbinate (Grassin-Delyle et al., Pharmacology & Therapeutics 134: 366-379(2012)). The nasal mucosa comprises an epithelial cell layer lining the nasal cavity. Drugs can be absorbed into the systemic circulation through the nasal mucosa.

[0064] Some exemplary nasal administration devices include steam inhalers, droppers, pipettes, squeeze bottles, spray pumps, nebulizers, powder sprays, and insufflators (Djupesland, Drug Deliv. And Transl. Res. 3:42-62, 2013). It should be understood that one skilled in the art can select an appropriate nasal administration device depending on the dosage form, chemical properties, physical properties, and other relevant considerations of a given pharmaceutical composition.

[0065] In some embodiments, the concentration of epinephrine in the pharmaceutical compositions described herein ranges from 0.3% to 5% (w / v). In some 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).

[0066] In some embodiments, the pharmaceutical compositions described herein are in the form of a liquid. In some embodiments, the pharmaceutical compositions described herein are in the form of a spray. In some embodiments, the pharmaceutical compositions form droplets.

[0067] permeability As used herein, "permeability" refers to the ability of a pharmaceutical composition to pass through a biofilm. For the purposes of this disclosure, the effective permeability coefficient (Pe) is used as an index of in vitro permeability testing in parallel artificial membrane permeability assays (PAMPA). The PAMPA permeability test is based on the passive diffusion of target compounds through an artificial membrane. The PAMPA artificial membrane has a lipid-oil-lipid sandwich structure constructed within the pores of a porous filter. The intermediate oil layer is ultrathin to maintain a strong and stable PAMPA membrane and minimize interference with compound retention and compound permeation. A test compound diluted with a buffer solution 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, a larger absolute Pe value indicates better in vitro permeability. The detailed procedure of PAMPA is described in Example 1.

[0068] The penetration enhancers disclosed in the present invention can effectively increase the permeability of epinephrine compositions. In one embodiment, the pharmaceutical compositions described herein exhibit a 3×10 permeability in the PAMPA assay. -6 It has a higher absolute value of Pe than cm / s.

[0069] In addition, when the pharmaceutical composition is administered to a subject, the maximum blood drug concentration (C max ) and C max Time to reach (T max ) are two useful indicators of in vivo permeability. max A larger value indicates better in vivo penetration, and T max A smaller value indicates better in vivo penetration.

[0070] Pharmacokinetic characteristics Compared with previously disclosed penetration enhancers, i.e., C12E9, n-dodecyl-β-D-maltoside (DDM), and diethylene glycol monoethyl ether (DEGEE), the penetration enhancers disclosed in the present invention exhibit more desirable pharmacokinetic characteristics. In certain embodiments, the pharmaceutical compositions disclosed in the present invention exhibit superior bioavailability. In certain embodiments, the pharmaceutical compositions disclosed in the present invention exhibit a faster absorption rate, i.e., a relatively short T max In one embodiment, the pharmaceutical composition disclosed in the present invention exhibits a larger initial area under the partial drug blood concentration-time curve, AUC.

[0071] The present disclosure further provides the unexpected discovery that, compared to previously disclosed penetration enhancers (e.g., C12E9, DDM, and DEGEE), the pharmaceutical compositions described in the present disclosure exhibit higher bioavailability, particularly at relatively high epinephrine doses that more closely resemble clinically used doses. Some pharmaceutical compositions described in the present disclosure may even exhibit higher bioavailability. For example, compared to intramuscular epinephrine, an epinephrine composition containing C12E7 as a penetration enhancer exhibits 2.7-fold higher exposure upon nasal administration.

[0072] Compared to previously disclosed compositions (e.g., C12E9, DDM, and DEGEE), the compositions disclosed in the present disclosure have shorter T max and a larger initial partial area under the drug blood concentration-time curve, AUC. Since epinephrine is a life-saving drug used in emergencies, it is highly desirable for epinephrine to act more quickly and effectively. max and a relatively large initial partial area under the drug blood concentration-time curve, AUC, are more advantageous characteristics (Assessment report of Neffy, EMA / 204348 / 2022, Committee for Medicinal Products for Human Use, 25 March 2022).

[0073] safety Compared to previously disclosed penetration enhancers (e.g., C12E9, DDM, and DEGEE), the penetration enhancers disclosed in the present disclosure have superior safety performance. In certain embodiments, the pharmaceutical compositions disclosed in the present disclosure are better tolerated. In certain embodiments, the pharmaceutical compositions disclosed in the present disclosure do not cause irreversible damage to the nasal mucosa.

[0074] Epinephrine has a relatively narrow therapeutic window. Doses exceeding this window can lead to potentially fatal side effects, such as cerebrovascular hemorrhage and subarachnoid hemorrhage. Therefore, an ideal penetration enhancer must enhance epinephrine tolerance. In a disclosed pharmacokinetic study, all subjects in the C12E9 group who received intranasal administration of an epinephrine composition containing C12E9 (1 mg / kg dose of epinephrine) died, demonstrating only a 53.2% bioavailability compared with intramuscular administration. However, after receiving intranasal administration of an epinephrine composition containing C12E7 (1 mg / kg dose of epinephrine), only one subject was in shock (recovered after cardiopulmonary resuscitation), and the bioavailability exceeded 270% compared with intramuscular administration. In contrast, after receiving intranasal administration of 1 mg / kg doses of epinephrine compositions containing C12E23 and C16E10, respectively, the activity of the subjects was significantly reduced, indicating some degree of adverse reaction. On the other hand, after receiving intranasal administration of epinephrine compositions containing C12E8, C12E4, or C10E6 (1 mg / kg doses of epinephrine), the subjects remained normal and active, and showed higher F values, shorter T max and a larger initial area under the partial drug blood concentration-time curve, AUC, is observed. The above observations indicate that C12E7, C12E8, C12E4, and C10E6 can unexpectedly enhance tolerance to epinephrine. When C12E7, C12E8, C12E4, and C10E6 are added to an epinephrine composition, the subject's tolerance to the composition can be enhanced. In one embodiment, the epinephrine composition is administered to a subject via the nasal route.

[0075] For purposes of this disclosure, a subject's better "tolerance" of a drug or pharmaceutical composition means that the subject has fewer adverse reactions, has a milder adverse reaction, or is less likely to experience an adverse reaction after administering the drug or pharmaceutical composition (e.g., epinephrine or a composition containing the epinephrine composition) to the subject. Tolerance describes the subject's ability to tolerate large amounts of an active ingredient (e.g., epinephrine).

[0076] As used herein, irreversible damage refers to damage that cannot be self-repaired in a relatively short time. Due to the strong local pharmacological effect of epinephrine, irreversible damage may be more severe when a penetration enhancer is used together with epinephrine. For the purposes of the present disclosure, an 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 strongly limits the absorption of drugs with a molecular weight greater than 1 kDa. Therefore, insulin with a molecular weight greater than 5 kDa generally cannot pass through the nasal mucosa unless the nasal mucosa is damaged. When insulin is administered nasally after nasal exposure to a penetration enhancer, the C of insulin after nasal administration of insulin at different times after nasal administration of a composition containing the drug (i.e., epinephrine) max By monitoring the C and AUC, it is possible to determine whether the composition causes any damage to the subject's nasal mucosa and whether such damage, if any, is reversible. max The C and AUC indicate relatively little damage to the nasal mucosa, while the C of relatively high levels of insulin exposuremax and AUC indicate the presence of damage to the nasal mucosa. In the case of reversible nasal mucosa damage, the C max The C and AUC of insulin are initially very high and then rapidly decrease, i.e., return to a significantly lower level within about 2 hours. In the case of irreversible nasal mucosal damage, max and AUC is maintained at a relatively high level for more than 2 hours. For detailed procedures of the insulin absorption test, see Example 5.

[0077] The present disclosure further provides the unexpected discovery that the pharmaceutical composition exhibits faster reversal of nasal mucosal damage at low pH (below 7, e.g., pH values ​​in the range of 4 to 6). When the pharmaceutical composition is formulated to have a pH below 7, the pharmaceutical composition causes only reversible damage, which can be repaired more quickly. In contrast, many of the permeation enhancers known in the art for use in intranasal formulations, such as DDM or DEGEE, do not exhibit the pH-sensitivity tendency toward nasal mucosal damage disclosed in Example 5.

[0078] In addition, the pharmaceutical composition has better stability and permeability under low pH conditions, ie, at pH values ​​below 7, for example, in the range of 4-6.

[0079] Even more surprisingly, compared to previously disclosed compositions (e.g., compositions containing a penetration enhancer such as C12E9, DDM, or DEGEE), the current compositions exhibit less nasal mucosal damage at low pH conditions, i.e., at pH values ​​less than 7, e.g., in the range of 4 to 6. For example, when epinephrine is administered via the nasal route in a composition having a pH of 7.4, C12E9 has been reported to cause severe nasal mucosal damage (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 the discovery that C12E9 causes much less nasal mucosal damage at low pH conditions, i.e., at pH values ​​less than 7, e.g., in the range of 4 to 6.

[0080] In some embodiments, the pharmaceutical composition has a pH value of less than 7. In some embodiments, the pharmaceutical composition has a pH value in the range of 4 to 6. In some embodiments, the pharmaceutical composition has a pH value of 4, 5, or 6.

[0081] The present disclosure further provides a pharmaceutical composition comprising epinephrine and C12E9 as a penetration enhancer, wherein the pharmaceutical composition has a pH value of less than 7, for example in the range of 4-6.

[0082] stability In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable auxiliary agent, wherein the pharmaceutically acceptable auxiliary agent is a pH adjusting agent, an antioxidant, a preservative, or an osmolality adjusting agent.

[0083] A pH adjuster, sometimes called a pH regulator or acidity regulator, is a substance for adjusting the pH of a pharmaceutical composition to a specific range. pH is an expression of the hydrogen ion concentration in water. Specifically, pH is the concentration of hydrogen ions (H +) is the negative logarithm of the concentration (mol / L), and pH = -log 10 (H + In certain embodiments, the pH adjusting agent is an acid or a base. Some exemplary pH adjusting agents include hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, and ammonia.

[0084] Antioxidants are compounds that inhibit oxidation. In some embodiments, antioxidants are used to improve the stability of pharmaceutical compositions by retarding the oxidation of active ingredients and other adjuvants. Some exemplary antioxidants and adjuvants include cysteine ​​(CYS), sodium metabisulfite (SMB), propyl gallate (PG), butylhydroxytoluene (BHT), D-α-tocopheryl polyethylene glycol succinate (vitamin E TPGS), ascorbic acid (VC), methionine, sodium bisulfite, sodium sulfite, α-lipoic acid, and butylhydroxyanisole (BHA) (Celestino et al., Brazilian J. Pharma. Sci. 43-3, 405-415 (2012)).

[0085] Preservatives are substances added to pharmaceutical compositions to prevent adverse physical, chemical, or biological changes. In some embodiments, the preservative is a bactericide or antibacterial agent. Some exemplary preservatives include benzalkonium chloride, trichlorotert-butanol, butylparaben, propylparaben, benzethonium chloride, chlorocresol, phenol, and benzoic acid.

[0086] Osmotic agents are generally water-soluble substances with small molecular weights. Osmosis is the diffusion of water under osmotic pressure due to an imbalance of molecules on both sides of a membrane. Osmotic agents can change the osmotic pressure across a cell membrane. Some exemplary osmotic agents include sodium chloride, glucose, mannitol, sorbitol, lactose, phosphoric acid, and citric acid.

[0087] In some embodiments, the pharmaceutical composition includes an antioxidant. In some embodiments, 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), butylhydroxytoluene (BHT), and butylhydroxyanisole (BHA). In some embodiments, the antioxidant is selected from sodium bisulfite and sodium metabisulfite. In some embodiments, the pharmaceutical composition includes 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), epinephrine formulations containing C10E6, C12E4, C12E7, C12E8, and C12E9 as penetration enhancers exhibit significantly better color stability characteristics. Pharmaceutical compositions comprising a penetration enhancer and an antioxidant have better stability and permeability at low pH conditions, ie, at pH values ​​below 7, such as in the range of 4-6.

[0088] As used herein, "stability" refers to the ability of a pharmaceutical composition to maintain its chemical, physical, and biological properties over time in the context of a pharmaceutical composition. For purposes of this disclosure, stability refers to the "percent content" of the active ingredient in the pharmaceutical composition. As used herein, the percent 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, where the composition is stored under the specified environmental conditions (pH, temperature) for the entire duration of the accelerated stability test, starting from day 0, and the percent content is in weight percent.

[0089] In one embodiment, the pharmaceutical composition has a content percentage of 90% or more after 30 days under conditions of pH 4.0 and 60°C.

[0090] method In another aspect, the present disclosure provides a method comprising administering to a subject an effective amount of the pharmaceutical composition described above. The route of administration may be, for example, intravenous, intratumoral, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, or ocular.

[0091] In one embodiment, the method is used to treat a subject suffering from a type I allergic reaction. A type I allergic reaction is an IgE-mediated immune response involving the release of antibodies against soluble antigens. This leads to the degranulation of mast cells 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., allergic shock, urticaria, angioedema, food, and drug allergies).

[0092] In certain embodiments, the allergic reaction is selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, allergic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

[0093] In certain embodiments, the pharmaceutical composition is administered to the subject by nasal delivery.

[0094] As used herein, "C max " refers to the maximum blood drug concentration shown in the time-course curve of the blood drug concentration of an active drug ingredient (e.g., epinephrine) or its metabolite. It should be understood that a person skilled in the art can select an appropriate method and conditions to measure the epinephrine blood drug concentration of a subject and obtain the C max It should be understood that a person skilled in the art can select an appropriate method and conditions to measure the epinephrine blood drug concentration of a subject and determine T max can be determined.

[0095] As used herein, AUC refers to the area under the blood drug concentration versus time curve, i.e., the area under the curve of the change in blood drug concentration of an active ingredient or its metabolite over time after administration of a certain dose of the active ingredient (e.g., epinephrine). "AUC 0-∞ ” is the area under the concentration-time curve extrapolated to infinity after administration, and “AUC 0-t " is the area under the concentration-time curve from zero to time t after administration, where t is the last time point of measurable concentration.

[0096] As used herein, bioavailability (F) describes the percentage of an administered dose of drug that reaches the systemic circulation. Its meaning is as defined in 21 CFR § 320.1(a). For purposes of this disclosure, F specifically refers to the ratio of unit dose exposure between intranasal and intramuscular routes of administration. For example, F(AUC 0-t )=(AUC 0-t,鼻内 / dose 鼻内 ) / (AUC 0-t,筋肉内注射 / dose 筋肉内注射 )×100%.

[0097] As used herein, initial fractional bioavailability (F 0-10 ) specifically refers to the ratio of AUC within the first 10 minutes after administration of a unit dose between the intranasal route and the intramuscular injection route. For example, F(AUC 0-10 )=(AUC 0-10,鼻内 / dose 鼻内 ) / (AUC 0-10,筋肉内注射 / dose 筋肉内注射 )×100%.

[0098] Example The present specification may be further illustrated by the following non-limiting examples, which may, where appropriate, employ standard techniques known to those skilled in the art and techniques similar to those illustrated in these examples, and it should be understood that those skilled in the art may envision other embodiments consistent with the description provided by the present invention.

[0099] Example 1 In vitro permeability evaluation of PAMPA The effects of different penetration enhancers on the in vitro permeability of epinephrine were evaluated using a 96-well PAMPA reagent kit (96-well skin PAMPA Sandwich set, PION Inc., MA, USA).

[0100] PRISMA TM The buffer solution is 25 mL of PRISMA TM (P / N 110151, PION Inc., MA, USA) was diluted to 1 L with ultrapure water and then adjusted to pH 4.0 with 0.5 M NaOH solution.

[0101] 200 μL of hydration solution (PION Inc., MA, USA) was added to each well of the supply tip (P / N 110660, PION Inc., MA, USA). Then, pre-coated PAMPA plates (P / N 120657, PION Inc., MA, USA) were immersed in the hydration solution overnight to allow for sufficient hydration.

[0102] The designed compositions were prepared by dissolving the permeation enhancer (at the concentrations listed in Table 2) together with epinephrine (10 mg / mL, 50 mg / mL, or 3 mg / mL) in a pH 4.0 aqueous buffer solution. A pH 4.0 epinephrine solution (without permeation enhancer) served as the control. 200 μL of each composition solution was accurately added to each well of the supply terminal, and 200 μL of PRISMA TM Buffer solution was added to each well of the receiving end. Next, a PAMPA artificial membrane was placed between the two plates (the supply end and the receiving end) to begin the in vitro permeability evaluation experiment. The permeation experiment for each composition was repeated in parallel in four wells. The PAMPA reagent kit containing the supply end, PAMPA artificial membrane, and receiving end was capped and incubated at 37°C for 5 hours. Additionally, an antioxidant was used to prevent oxidation during the experimental process. After incubation, the epinephrine concentrations in the supply end and receiving end solutions were measured by UPLC using the method described in Table 1.

[0103] [Table 1]

[0104] The polyoxyethylene alkyl ethers containing a double bond in the alkyl chain below were designated C18-1E10 and C18-1E20. Table 2 shows the PAMPA test results for epinephrine solutions containing different types of penetration enhancers (0.1%, 0.25%, 2.5% w / v or v / v). The negative control (epinephrine solution without penetration enhancer, only 1.1% penetrated the receptor end, with a Pe value of 0.45 x 10) -6 Compared with the 0.1% to 2.5% epinephrine solutions containing C10E6, C12E7, C12E8, C12E9, and C16E10, the epinephrine solutions all showed significantly more consistent epinephrine concentrations in the receiving end solution and higher Pe values, i.e., more than 8% epinephrine penetrated the receiving end, resulting in Pe values ​​of 3 × 10 -6 Compared to DEGEE, all PAMPA in vitro permeability studies showed significantly improved permeability for compositions containing 0.1% to 2.5% C10E6, C12E7, C12E8, C12E9, and C16E10, with some permeation enhancers exhibiting higher permeability than DDM, which is more clinically significant, especially when the permeation enhancer concentration is relatively low.

[0105] [Table 2] JPEG2025535459000004.jpg124169

[0106] Example 2 Pharmacokinetic (PK) Study of a Composition Containing a Penetration Enhancer Following Nasal Administration The preparation methods for different formulations for intranasal administration were as follows: First, an aqueous solution containing 0.25% penetration enhancer, 0.2% sodium bisulfite, and 0.9% sodium chloride was prepared, followed by dissolving epinephrine to a concentration of 10 mg / mL or 3 mg / mL. All formulations were then adjusted to pH 4 and refrigerated. Liquid compositions containing epinephrine and penetration enhancer were administered nasally according to the method in Table 3.

[0107] The prepared solution compositions were intranasally administered to Sprague-Dawley (SD) rats, and blood samples were collected in 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90, and 120 minutes after administration. A 10% (w / v) aqueous solution of sodium pyrosulfite was prepared and added to the samples at a volume ratio of 9:1 to prevent oxidation. The samples were thoroughly vortexed and stored in an ice-water bath until further processing. If sample preparation and analysis were not performed on the day of the PK experiment, the samples were stored in a -80°C refrigerator. Frozen samples were allowed to equilibrate to room temperature before further processing. After thawing the samples, 20 μL of plasma sample was transferred to a 1.5 mL polyethylene centrifuge tube and added to 180 μL of internal standard solution (see Table 5 for preparation method). The mixed sample was vortexed for 5 minutes and centrifuged at 10,000 rpm at 4°C for 5 minutes. 130 μL of the supernatant was collected into a 96-well plate, and 130 μL of ultrapure water was added. The plate was then vortex-mixed for 5 minutes and centrifuged 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.

[0108] [Table 3]

[0109] [Table 4]

[0110] [Table 5]

[0111] [Table 6]

[0112] [Table 7]

[0113] The time course of plasma epinephrine concentrations in the 10 mg / mL epinephrine concentration formulation (1 mg / kg dose) group is shown in Table 8 and Figure 1. In the intramuscular injection PK study, three SD rats were used in each group, with an epinephrine concentration of 1 mg / mL and a dose of 0.1 mg / kg. In the nasal administration PK study, two SD rats were used in each group, with an epinephrine concentration of 10 mg / mL and a dose of 1 mg / kg. In Examples 2 and 3, "NA" indicates that epinephrine was not detected in plasma at a particular time point, or that the parameter could not be calculated because epinephrine was not detected at a particular time point. As shown in Table 9, the PK parameters, i.e., T max , C max , AUC, bioavailability (F 0-t ) and the initial fractional bioavailability (F) within the first 10 minutes after administration 0-10 ) was calculated using MaS Studio (v1.5.3.10). From observations, when the polyoxyethylene alkyl ether has 9 to 15 methylene groups and 4 to 10 oxyethylene units, C max , AUC and bioavailability were found to be significantly higher.

[0114] [Table 8]

[0115] [Table 9]

[0116] The time course of plasma epinephrine concentration in the 3 mg / mL epinephrine concentration formulation group (much lower than the clinically used 0.3 mg / kg dose) is shown in Table 10 and Figure 2. As shown in Table 11, the PK parameters, i.e., T max , C maxThe AUC and bioavailability were calculated using MaS Studio (v1.5.3.10). From the observation, when the polyoxyethylene alkyl ether has 9-15 methylene groups and 4-10 oxyethylene units, T max is shorter and at the same time has a relatively high C max From a clinical point of view, epinephrine is required to act more rapidly for the treatment of type I allergic reactions, so a relatively short T max is more ideal.

[0117] [Table 10]

[0118] [Table 11]

[0119] Example 3 Pharmacokinetic (PK) Study of Compositions with Different Penetration Enhancer and Epinephrine Concentrations The process for preparing compositions for intranasal administration was as follows: First, aqueous solutions containing 0.1%, 0.25%, 1.0%, and 2.5% penetration enhancer, 0.2% sodium bisulfite, and 0.9% sodium chloride were prepared. Epinephrine was then dissolved to a drug concentration of 10 mg / mL or 3 mg / mL (except for the 2.5% DEGEE composition, which had an epinephrine concentration of 25 mg / mL). The resulting solutions were adjusted to pH 4 and refrigerated. Liquid compositions containing epinephrine and penetration enhancer were administered nasally according to the method shown in Table 12.

[0120] Different formulations were instilled into the nasal cavities of SD rats, and blood samples were collected in 1.5 mL polyethylene centrifuge tubes at 5, 10, 15, 30, 45, 60, 90, and 120 minutes after administration. A 10% (w / v) aqueous solution of sodium pyrosulfite was prepared and added to the samples at a volume ratio of 9:1 to prevent oxidation. The samples were vortexed thoroughly and stored in an ice-water bath until further processing. If samples were not pretreated and analyzed on the day of the PK experiment, they were stored in a refrigerator at -80°C. Frozen samples were allowed to equilibrate to room temperature before further processing. After thawing, 20 μL of plasma sample was transferred to a 1.5 mL polyethylene centrifuge tube and added to 180 μL of internal standard solution (see Table 5 for preparation method). The mixed sample was vortexed for 5 minutes and centrifuged at 10,000 rpm at 4°C for 5 minutes. 130 μL of the supernatant was collected into a 96-well plate, and 130 μL of ultrapure water was added. The plate was then vortexed for 5 minutes and centrifuged 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.

[0121] [Table 12]

[0122] The plasma epinephrine concentrations at different time points after administration of compositions with different epinephrine concentrations and penetration enhancer types and concentrations are shown in Table 13 and Figure 3. It was observed that the tested polyoxyethylene alkyl ethers had obvious penetration enhancing ability in the concentration range of 0.1 mg / mL to 2.5 mg / mL. As shown in Table 14, the PK parameters, i.e., T max , C max , AUC, F 0-t and F 0-10 was calculated using MaS Studio (v1.5.3.10). When the administration volume is constant, C increases as the penetration enhancer concentration and epinephrine concentration increase. maxThe results showed that the AUC and bioavailability generally increased. Compared with the groups containing DDM and DEGEE, at the same penetration enhancer and epinephrine concentrations, the polyoxyethylene alkyl ethers both showed higher C max , AUC and bioavailability, which suggests its better permeation enhancing ability. The formulation containing polyoxyethylene alkyl ether showed a shorter T max was further observed, indicating that epinephrine takes effect more rapidly.

[0123] [Table 13]

[0124] [Table 14]

[0125] Example 4 Pharmacokinetic (PK) Study of a Composition Containing a Penetration Enhancer Following Nasal Administration The different formulations for intranasal administration were prepared as follows: First, an aqueous solution containing 0.25% penetration enhancer, 0.2% sodium metabisulfite, and 0.9% sodium chloride was prepared, followed by dissolving epinephrine to an epinephrine concentration of 2.5 mg / mL, 10 mg / mL, or 20 mg / mL. All formulations were then adjusted to pH 4 and refrigerated. Liquid compositions containing epinephrine and penetration enhancer were administered nasally to beagle dogs according to the method in Table 15.

[0126] The composition was administered intranasally to beagle dogs, and blood samples were collected 1, 5, 10, 15, 20, 30, 60, 90, and 120 minutes after administration. Whole blood samples were collected in 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. Samples to be analyzed were thawed and vortexed at room temperature. A 100 μL sample was taken, and 20 μL of water (containing 0.1% acetic acid and 50 ng / mL epinephrine-D6) was added as a protein precipitant. 250 μL of PBA was then added, vortexed to homogenize, and 400 μL of TOAB was added. The mixture was vortexed for 10 minutes and then centrifuged at 13,000 rpm for 5 minutes. 300 μL of the supernatant was collected in a 1.5 mL centrifuge tube, and 200 μL of n-octanol and 125 μL of 0.02 N aqueous hydrochloric acid were added and vortexed for 3 min. The lower layer solution was removed and mixed with 50 μL of sodium tetraborate buffer salt (100 mmol) and 100 μL of benzoyl chloride (1%) and vortexed for 0.5 min. 10 μL of the final mixed sample solution was injected into an LC-MS / MS system for analysis according to the methods listed in Table 16 and Table 7, respectively.

[0127] [Table 15]

[0128] [Table 16]

[0129] Table 17 and Figure 4 show the time course of plasma epinephrine concentrations at different time points after administration of the formulations with different dosage concentrations and types of penetration enhancers. As shown in Table 18, the corresponding pharmacokinetic parameters (i.e., T max , C max , AUC, bioavailability F for intramuscular injection route 0-t and the initial fractional bioavailability F within the first 10 minutes after administration 0-10) was calculated using Phoenix WinNonlin 8.3 software. Compared with commercially available formulations using DDM as a penetration enhancer, the C12E7 group still exhibited higher bioavailability, even at lower epinephrine concentrations and doses. The C12E7 group also exhibited an approximately three-fold higher early partial AUC compared with the DDM group (the currently used commercially available formulation). Of note, a relatively high early partial AUC is highly desirable when using epinephrine to treat type I allergic reactions (including allergic shock) in emergency situations.

[0130] [Table 17]

[0131] [Table 18]

[0132] Example 5 Evaluation of the stability of the composition Epinephrine was dissolved in an aqueous solution containing 0.25% of different penetration enhancers and 0.9% sodium chloride to prepare intranasal compositions with an epinephrine concentration of 10 mg / mL. To prepare formulations containing antioxidants and preservatives, 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 formulations containing antioxidants at 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 at pH 7.4 containing 0.2% antioxidant and phosphate buffered saline (PBS) was also prepared. The stability test design for the compositions is shown in Table 19.

[0133] [Table 19]

[0134] The liquid composition was dispensed into vials, which were then tightly capped and placed in an oven for accelerated stability testing. <391> Based on the Epinephrine Assay, the epinephrine content in different compositions was quantified using high performance liquid chromatography (HPLC) before and after the accelerated stability test, and the analytical method is shown in Table 20. The preparation method of phosphate buffer (pH 2.8) is as follows: 5.0 g / L potassium dihydrogen phosphate and 2.6 g / L sodium octanesulfonate were added to water, and then the pH was adjusted to 2.8. The phosphate buffer was filtered through a 0.45 μm filter before use.

[0135] [Table 20]

[0136] The stability test results of compositions containing epinephrine and different penetration enhancers after accelerated storage at 60°C for different periods are shown in Table 21. All formulations were observed to have a significant decrease in epinephrine content after acceleration. C12E9 and C16E2 exhibited relatively poor stability characteristics compared to the other compositions. The remaining compositions exhibited similar stability characteristics.

[0137] [Table 21]

[0138] The stability test results (at 60°C for different time periods) of compositions containing 10 mg / mL epinephrine, 0.25% C12E8 or DDM, and antioxidants, chelating agents, and preservatives are shown in Table 22. Sodium bisulfite significantly improved the stability of the compositions, and when sodium bisulfite and EDTA-2Na were used together, the appearance of the compositions remained clear for a longer period of time, while the compositions containing sodium bisulfite exhibited a red to dark brown color after acceleration.

[0139] [Table 22]

[0140] The stability test results after accelerated testing at 60°C for 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 are shown in Tables 23 and 24. When the two preservatives were added simultaneously, no significant difference in stability was observed, indicating good chemical compatibility.

[0141] [Table 23]

[0142] [Table 24]

[0143] The stability test results for 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 accelerated incubation at 60°C are shown in Table 25. The antioxidant sodium bisulfite significantly improved the stability of the compositions in the pH range of 4.0 to 6.0, and after 7 days of accelerated incubation at 60°C, the epinephrine content remained above 95%. However, at pH values ​​above 7.0, the stability of the compositions rapidly decreased. It was also observed that sodium bisulfite improved the stability of the compositions only at pH 4.0 and 6.0, and that the addition of sodium bisulfite further reduced the stability of the compositions at pH values ​​above 7.0.

[0144] [Table 25]

[0145] Example 6 Evaluation of nasal mucosal damage after nasal administration of an epinephrine-containing composition Because both permeation enhancers and epinephrine cause nasal mucosal damage, a study was designed and conducted to evaluate nasal mucosal damage after nasal administration of a composition containing epinephrine and different permeation enhancers. Human insulin is poorly absorbed through the nasal mucosa unless the nasal mucosa is damaged. Therefore, as described in a literature review (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 evaluated by measuring insulin absorption through the nasal mucosa at different times after nasal administration of a composition containing a permeation enhancer and epinephrine. A relatively high level of insulin absorption through the nasal cavity indicated nasal mucosal damage, while a lack of insulin absorption indicated intact nasal mucosa.

[0146] The evaluation method was as follows: First, liquid compositions containing a penetration enhancer and epinephrine were prepared according to Table 26, and a 0.5 μU / mL normal human insulin solution was also prepared. Eight healthy SD rats were used as the test group for each composition, and two SD rats were used for each subgroup at each time point for each composition. Then, as shown in Table 22, each composition was administered intranasally (dosage: 0.1 mL / kg) to the rats in all four corresponding subgroups at each time point. In the 0 hour subgroup, SD rats were administered regular human insulin (0.05 μU / kg dose, 0.5 μU / mL concentration, 0.1 mL / kg dose) intranasally immediately after administration of the epinephrine composition; in the 2 hour subgroup, SD rats were administered regular human insulin (0.05 μU / kg dose, 0.5 μU / mL concentration, 0.1 mL / kg dose) intranasally 2 hours after administration of the epinephrine composition; in the 4 hour subgroup, SD rats were administered regular human insulin (0.05 μU / kg dose, 0.5 μU / mL concentration, 0.1 mL / kg dose) intranasally 4 hours after administration of the epinephrine composition; and in the 8 hour subgroup, SD rats were administered regular human insulin (0.05 μU / kg dose, 0.5 μU / mL concentration, 0.1 mL / kg dose) intranasally 8 hours after administration of the epinephrine composition. For all rats, blood samples were collected at 0, 10, 20, 30, 45, 60, 90 and 120 minutes after administration of regular human insulin, and then plasma insulin concentrations were measured using an ELISA reagent kit (Mercodia brand, operated according to the instruction manual), and then the PK parameters of each subgroup were calculated using MaS Studio software (v1.5.3.10) (Table 27, Figure 5 and Figure 6), thereby reflecting the time-dependent changes in nasal mucosa damage after administration of different compositions.

[0147] [Table 26] JPEG2025535459000029.jpg238169JPEG2025535459000030.jpg48169

[0148] Surprisingly, the pharmacokinetic parameters of insulin showed that the composition containing C12E9 showed irreversible damage at a relatively high pH value (e.g., pH 7.4), while relatively mild and reversible damage was observed when the pH of the composition was relatively low (e.g., pH 4.0). Compared to the irreversible nasal mucosa damage observed using DDM, DEGEE, and C12E9 (high pH), all of the compositions containing polyoxyethylene alkyl ethers disclosed in the present application showed that the nasal mucosa recovered to a normal state at a relatively fast rate (within 2 hours) at a relatively low pH. In addition, the tested compositions containing polyoxyethylene alkyl ethers showed high insulin AUC and C at 0 hours. max It was also observed that the polyoxyethylene alkyl ethers exhibited a pH of 1.0 or higher, which indicated faster drug absorption compared to DDM, DEGEE, and C12E9 (high pH). Therefore, when used in an appropriate pH environment, the polyoxyethylene alkyl ethers disclosed in the present application can be used as safer (e.g., faster recovery of the nasal mucosa) and more effective (faster drug absorption after administration) penetration enhancers compared to other penetration enhancers disclosed in the prior art.

[0149] [Table 27]

Claims

1. Epinephrine and the formula CH 3 (CH 2 ) n-1 [OCH 2 CH 2 ] m and a penetration enhancer having an OH group, 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; Pharmaceutical compositions.

2. n is 12 and m is selected from 4, 7 and 8; The pharmaceutical composition of claim 1.

3. n is 10 and m is 6, The pharmaceutical composition of claim 1.

4. The concentration range of the penetration enhancer is 0.1% to 2.50% (v / v); The pharmaceutical composition according to any one of claims 1 to 3.

5. The concentration of the penetration enhancer is 0.25% (v / v). The pharmaceutical composition according to any one of claims 1 to 4.

6. The pH value of the pharmaceutical composition is less than 7. The pharmaceutical composition according to any one of claims 1 to 5.

7. The pH range of the pharmaceutical composition is 4 to 6. The pharmaceutical composition according to any one of claims 1 to 6.

8. The pharmaceutical composition is used for nasal administration or for use with a nasal administration device. The pharmaceutical composition according to any one of claims 1 to 7.

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

10. 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), butylhydroxytoluene (BHT), and butylhydroxyanisole (BHA); The pharmaceutical composition of claim 9.

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

12. The concentration range of epinephrine in the pharmaceutical composition is 0.3% to 5% (w / v). The pharmaceutical composition according to any one of claims 1 to 11.

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

14. The pharmaceutical composition has a recovery rate of 90% by weight or more after 30 days under conditions of pH 4.0 and 60°C. The pharmaceutical composition according to any one of claims 1 to 13.

15. The pharmaceutical composition has an absolute Pe of 3×10 in the PAMPA test. -6 More than cm / s The pharmaceutical composition according to any one of claims 1 to 14.

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

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

18. The method is used to treat a subject suffering from a type I allergic reaction.

18. The method of claim 17.

19. The method is used to treat a condition selected from allergic asthma, allergic conjunctivitis, allergic rhinitis, allergic shock, angioedema, urticaria, eosinophilia, drug allergy, and food allergy.

18. The method of claim 17.

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

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