CN118141792A

CN118141792A - Compositions comprising epinephrine and prodrugs thereof

Info

Publication number: CN118141792A
Application number: CN202211549747.3A
Authority: CN
Inventors: 梁文飞; 程幸运
Original assignee: Guangzhou Dazhou Biomedical Technology Co ltd
Current assignee: Guangzhou Dazhou Biomedical Technology Co ltd
Priority date: 2022-12-05
Filing date: 2022-12-05
Publication date: 2024-06-07

Abstract

The invention provides a composition containing epinephrine and prodrug thereof, and relates to the technical field of pharmaceutical preparations. Sublingual and nasal routes of administration are non-invasive and are preferred over injectable administration. Due to the high vascularization of the mucosa, the drug can be rapidly absorbed through the mucosa and directly enter the systemic circulation, thereby avoiding longer gastrointestinal transit times and gastrointestinal and hepatic first pass effects due to metabolism. The two administration routes can meet the requirement of quick effect of the medicine required by anaphylactic reaction. Absorption of the drug through the sublingual or nasal cavity requires penetration of the drug into mucosal tissue. The composition containing epinephrine and the prodrug thereof provided by the invention can quickly increase the epinephrine concentration to a proper effective treatment concentration through a permeation enhancement technology, and maintain the concentration for a period of time, so that adverse reactions caused by the ultra-high peak concentration are avoided, and more convenient and effective treatment selection is brought to allergic patients.

Description

Compositions comprising epinephrine and prodrugs thereof

Technical Field

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a composition containing epinephrine and a prodrug thereof.

Background

Anaphylaxis is a systemic and life threatening anaphylactic reaction that produces a characteristic reactive anaphylactic shock, often accompanied by a decrease in blood pressure and consciousness. Allergic reactions may be triggered by a variety of allergens, including but not limited to insect bites, foods, drugs, allergens, etc. (Simons et al, 2010). The lifetime risk of allergy is 1% to 3% and mortality is 1% (Kemp et al, 2008).

Epinephrine is an endogenous catecholamine, mainly produced by norepinephrine in the adrenal medulla. Epinephrine is an alpha and beta adrenergic receptor agonist that directly acts on sympathomimetic nerves, with complex, adenosine-phosphate-mediated, bi-directional pharmacological effects on many target organs (Simons et al, 2004). Epinephrine was first approved by the U.S. food and drug administration in 1939 and has been used extensively, particularly in the treatment of allergic reactions. It is suggested as a first line of choice for allergy treatment (Simons et al, 2010).

Early and timely administration of epinephrine is critical to the treatment of allergic reactions, as the fatal allergic reactions of humans occur within minutes, and later administration may be futile (Sampson et al, 1992). In China, epinephrine is currently only available as an injection, and there is no product of the automatic injection device class. The specific use mode is that the epinephrine powder injection is stored by adopting an ampoule, and when in use, medical staff prepare injection for injection. In the recommendation of the severe allergic reaction first aid guide written by Li Xiaotong et al, the epinephrine is recommended to be given according to 0.01mg/kg body weight, the single maximum dose of patients aged 14 years and older is not more than 0.5mg, and the single maximum dose of patients aged 14 years and older is not more than 0.3mg; the concentration was 1mg/ml (1:1000) (Li Xiaotong et al, 2019). Because of the absence of automatic injection devices, allergic shock of some domestic patients is difficult to obtain epinephrine for first time for emergency treatment, and even death is caused in severe cases. Moreover, a questionnaire performed by s & n et al on 50 doctors showed that 34% of clinicians and 42% of nurses were first injected subcutaneously in the treatment of anaphylactic shock by epinephrine, rather than intramuscular injection first in the recommendation of the guidelines for severe anaphylaxis, 70% of clinicians and 76% of nurses considered that single doses of epinephrine would be 0.5-1.0mg, and 0.01mg/kg in the recommendation, or that single doses were no more than 0.5mg biased (s & n et al, 2021). When epinephrine is used for treating anaphylactic shock in China, the epinephrine not only has an automatic injector product which is not carried about, but also has the condition that the injection is complex in use process and not standardized in clinical training.

The conditions in the united states and europe are different from those in china, which treats acute allergic reactions, mostly using commercial auto-injector configurationsAuvi-Q, etc.), the auto-injection configuration occupies more than 90% of the epinephrine market share.

Notably, the pharmacokinetic profile of epinephrine is bimodal, whether by Epipen, auto-injector or intramuscular injection, with peak times Tmax1 and Tmax2 between 5-10 minutes and 30-50 minutes, respectively, as shown in fig. 1 (Drebog et al, ALLERGY ASTHMA CLIN Immunol, 2021).

It is particularly important that the medicaments for the first aid of the allergic reaction should be convenient and effective. Although guidelines in the united states, europe, etc. use epinephrine as a recommended drug for the treatment of acute allergic reactions, there is a serious lack of understanding of the use of epinephrine for the treatment of allergic reactions in China. In a survey report of tertiary hospitals, none of 225 allergic patients could acquire and use epinephrine at home, and the proportion of epinephrine used in emergency departments was only 9.3% (ginger et al, journal of allergic tissues worldwide, 2021). In another investigation, retrospective analysis was performed on outpatients diagnosed with allergic reaction or severe allergic reaction in the department of allergy in Beijing co-ordinates hospital from 1.2000 to 6.2014, 907 cases of the allergic reaction in 1952 were collected and analyzed, and 78% of adults and 22% of children were included in 907 patients. Of patients who received first aid, only 25% received epinephrine treatment (ginger et al, allergic asthma immune studies, 2016). For such life threatening diseases, the rate of epinephrine usage is too low, possibly due to the lack of convenient, economical epinephrine medication.

Sublingual administration is a rapid onset, non-invasive, simple administration. For example, the nitroglycerin sublingual tablet can be used for angina pectoris acute attack, and can be used for 5-15 minutes. The development into sublingual film agent is a brand new thought of epinephrine administration. Aqusitive Therapeutics, inc (nyquisfura medical Co., ltd.) patents US17/549,219 which disclose epinephrine or an epinephrine prodrug dipivefrin and adrenergic receptor compositions, which may further contain permeation enhancers. Based on the patent, AQST-108 sublingual film agent developed by the company adopts epinephrine prodrug dipivefrin as raw material, and enters a clinical second phase, but the conversion speed of the prodrug into epinephrine is not fast enough, the project of the prodrug is put aside after the second phase, and the development of the prodrug with faster conversion is turned to. The second generation product AQST-109 sublingual film shows faster absorption and conversion. However, AQST-109, although currently performing a second phase clinical trial, had significant differences in their pharmacokinetics and intramuscular injection. AQST-109 only one phase of absorption, the drug concentration drops rapidly after absorption, resulting in a relatively low drug exposure after 1 hour (Aquestive publication, 2021, see fig. 2). As described above, intramuscular injection resulted in a bimodal drug time profile, and the time required for intramuscular injection to reach a concentration of 100pg/mL in a Aquestive clinical trial was substantially similar to AQST-109, approximately 7-8 minutes, but intramuscular injection reached 100pg/L into a small plateau, and the drug concentration did not rise rapidly but maintained a relatively gentle rise.

The bimodal appearance of the epinephrine injection is caused by the vasoconstriction effect of epinephrine, and the epinephrine can generate the vasoconstriction effect after reaching a certain concentration, thereby reducing the absorption of epinephrine from muscles to blood vessels and avoiding adverse reaction caused by the overload of epinephrine. In comparison, AQST-109 reached peak much faster than intramuscular injection, but also eliminated much faster, which resulted in a much lower drug concentration than intramuscular injection after 1 hour, which is likely to lead to secondary allergic reactions. Thus, the route of AQST-109 prodrug alone remains a problem, which is not addressed by prodrugs alone, while allowing rapid absorption of epinephrine and at the same time extending drug exposure.

Intranasal injection of epinephrine is another potential method, and generally, nasal absorption is faster, for example, new Kang Taike (xylometazoline hydrochloride nasal spray) is used for symptoms such as rhinitis, and nasal obstruction can be relieved in about 2 minutes. However, nasal administration is difficult to maintain exposure to epinephrine over a prolonged period of time. As described above, nasal spray can solve the problem of rapid absorption of epinephrine due to the systemic half-life of epinephrine of only about 5 minutes, but cannot generate long-term drug exposure, and cannot simulate the drug time profile of injection, which can adversely affect adverse reaction and drug duration. In patent US10576156B2, a nasal spray composition is claimed, the patent comprising epinephrine, 0.05% -0.5% (w/v) alkyl glycoside, the patent further comprising citric acid, sodium citrate, propylene glycol, glycerol, ascorbic acid, sodium metabisulfite, disodium edetate, benzalkonium chloride, sodium hydroxide, or a combination thereof. The patent claims mainly comprise alkyl glycoside as nose absorption penetration enhancer, which can increase peak concentration by about 2 times and shorten peak time by about half. The patent does not mention any use of epinephrine prodrugs to improve nasal spray absorption and drug exposure profiles.

In general, in China, there is an urgent need for a more convenient, easy to use, non-invasive epinephrine product that helps allergic patients to rescue more quickly and effectively, and that also encourages people to increase their cognition for use in allergic emergency applications. In addition, in such products, the rapid absorption and prolonged drug exposure profile of epinephrine plays a critical role in the therapeutic efficacy of the drug.

Disclosure of Invention

The present invention addresses the problems of the prior art by providing a composition comprising epinephrine and a prodrug thereof. The composition comprising epinephrine and a prodrug thereof enables the concentration of epinephrine to be rapidly increased to a proper effective treatment concentration through a permeation enhancement technology, maintains the concentration for a period of time, avoids adverse reactions caused by the ultra-high peak concentration, and brings more convenient and effective treatment selection for allergic patients.

Certain embodiments of the present disclosure relate to methods of using epinephrine and prodrugs thereof to a subject in need thereof. In some embodiments, the method is for treating a disease or disorder in a subject in which epinephrine or a prodrug thereof is beneficial. In some embodiments, the method is for treating allergy (type I) including allergic reactions in a subject. Other indications include, but are not limited to, (1) those who have previously developed known food allergy-induced moderate-severe allergic reactions in specific parts of the body (e.g., peanut and/or tree nuts, milk, seafood, associated with severe reaction sites of predisposition); (2) The prior teenagers with mild-moderate reactions and young food allergy patients; (3) In the past, people who are exposed to food, hymenoptera insect toxins, latex or airborne allergens induce light-to-moderate allergic reactions, are far away from medical institutions, or need long-term travel; (4) In the past, trace foods induce light-medium allergic reaction; (5) Hymenoptera venom or drug allergy sufferers, symptoms are not limited to skin mucosa system responders and those with cardiovascular disease; (6) oral immunotherapy of food allergy. According to the following: severe allergic reactions are known adverse reactions of oral immunotherapy for food allergic patients.

In some embodiments, the method comprises administering to the subject a formulation of a combination of both epinephrine and an epinephrine prodrug. In some embodiments, the method comprises injecting the drug solution into the patient through the nasal cavity, sublingual mucosa, or muscle. In some embodiments, sublingual, nasal spray, intramuscular injection formulations using epinephrine and/or a prodrug thereof provide an effective concentration of epinephrine.

Generally, for any of the methods described above, a formulation of a combination of both epinephrine and an epinephrine prodrug is employed to provide a therapeutically effective concentration of epinephrine in the subject's plasma for a sustained period of time. Therapeutically effective concentrations of epinephrine may vary, typically between 100pg/mL and 2000pg/mL; preferably above 100pg/mL and below 2000pg/mL; preferably above 200pg/mL and below 1000pg/mL. The time for epinephrine to reach and maintain therapeutic concentration may be from 5 minutes to 6 hours, or 5 minutes to 2 hours, or 5 minutes to 1 hour.

In some embodiments, the formulation using a combination of both epinephrine and an epinephrine prodrug, including sublingual film, nasal spray, or intramuscular injection, allows the epinephrine concentration to be rapidly brought to the effective concentration for the treatment and maintained at the effective concentration for an extended period of time. The rapidly rising and sustained epinephrine concentration is critical for the treatment of life threatening allergic reactions. For example, the present invention provides substantial advantages over methods employing epinephrine alone or epinephrine prodrugs alone. When epinephrine is used alone, a rapid rise in effective concentration can be provided, but the half-life of epinephrine is only about 5 minutes, so epinephrine alone does not provide a prolonged drug concentration, and thus continues to provide efficacy. According to some studies, it has been shown that after a first rescue of allergy using epinephrine, about 20% of the probability of second-stage allergic reactions occurs, partly due to rapid elimination of epinephrine, resulting in a concentration drop below the effective therapeutic window. In general, an epinephrine auto-injector device commonly used abroad is composed of two injection devices, in order to prevent the phenomenon that a dose administered once cannot provide sufficient therapeutic effect or second-stage allergic reaction occurs. When the prodrug dipivefoli Lin Zhibei film former is used alone, as shown in fig. 18B of patent US20210128511A1, the concentration of the initial epinephrine is low when the formulation is used alone, because of the slow rate of conversion of the prodrug to epinephrine. The formulation begins to climb slowly after about 15 minutes, which is a significant problem for the first aid of allergic reactions. In contrast, the parent drug epinephrine generally reaches an effective concentration after injection administration for 5-10 minutes or even less.

In some embodiments, using a sublingual film, nasal spray or intramuscular injection of a combination of both epinephrine and an epinephrine prodrug of the present invention as a starting material allows for rapid achievement of therapeutically effective levels of epinephrine and for prolonged periods of time. In the first stage of the invention, the effect of rapid onset of action is achieved by epinephrine and its prodrug being permeated and converted within about 5 minutes during the rapid ascent stage of epinephrine. The second stage provides a second peak concentration of epinephrine and maintains an effective concentration for a longer period of time. For example, in some embodiments, the concentration of epinephrine is maintained at a concentration of 100pg/mL-2000pg/mL for approximately 5 minutes-120 minutes. In contrast, when epinephrine is used alone, this concentration can only be maintained for 5-60 minutes. The second stage results because the prodrug gradually and slowly converts to epinephrine after passing through mucosal tissue and entering the systemic circulation.

In some embodiments, the time for the epinephrine concentration to ramp up rapidly to an effective concentration may vary from 5 minutes to 30 minutes, preferably 5 to 10 minutes, during the first stage. The second stage may or may not provide a distinct second peak, but the second stage may provide a drug exposure time above the effective concentration of more than 6 hours, at least up to 1 hour. The effective concentration of epinephrine may vary, typically being higher than 100pg/mL and lower than 2000pg/mL. Preferably between 200pg/mL and 1000 pg/mL.

In various embodiments, the first stage achieves therapeutic effect in the concentration of epinephrine, primarily due to the uptake of epinephrine, or the immediate conversion of prodrug to epinephrine contribution following uptake. The prodrug, upon penetrating the tissue, and upon entering the blood circulation, may rapidly convert a portion to epinephrine, which may occur within 5-15 minutes, preferably equal to or less than 5 minutes. The second prolonged phase of maintenance of a therapeutically effective concentration of epinephrine is mainly due to slow absorption or slow conversion of the prodrug conversion, which may occur and last for more than 6 hours, 4 hours, 2 hours, at least 1 hour. The therapeutically effective concentration of epinephrine is then maintained for more than 6 hours, 4 hours, 2 hours, at least 1 hour.

In various embodiments, the combination of epinephrine and epinephrine prodrug may be split into combinations of different ratios. In some embodiments, the epinephrine/prodrug weight ratio may be from 0.1 to 10. Depending on the actual bioavailability of the two components and the rate of prodrug conversion.

In various embodiments, the combination of epinephrine and epinephrine prodrug may be combined in different weights. In practice, this depends on the actual bioavailability of the two components and the rate of prodrug conversion.

Generally, the pharmaceutical composition may comprise a polymeric film-forming matrix, epinephrine or an epinephrine prodrug, or a combination of both, in the polymeric matrix, and a permeation enhancer. In some embodiments, the pharmaceutical composition may further comprise a pH adjuster. In some embodiments, the pharmaceutical composition may further comprise a stabilizer.

In some embodiments, the penetration enhancer may be a glyceride. In some cases, the glyceride may be caprylic capric polyethylene glycol glyceride, oleoyl polyoxyethylene glyceride, glycerol monooleate.

In some embodiments, the penetration enhancer may be a plant extract essential oil. In some cases, the plant essential oil may be clove oil, peppermint oil, angelica oil.

In some embodiments, the penetration enhancer may be a fatty acid. In some cases, the fatty acid may be oleic acid, levulinic acid.

In some embodiments, the penetration enhancer may be an ester of oleic acid. In some cases, the ester of oleic acid may be oleyl oleate, glycerol monooleate, glycerol trioleate.

In some embodiments, the permeation enhancer may be diethylene glycol monoethyl ether.

In some embodiments, the penetration enhancer may be dimethyl sulfoxide.

In some embodiments, the pharmaceutical composition may be a film further comprising a polymer matrix, the pharmaceutically active ingredient and the permeation enhancer being contained in the polymer matrix.

In some embodiments, the pH adjuster may be citric acid, hydrochloric acid. In some embodiments, the pharmaceutical composition may have a pH of 1.5, 3.0, 5.0, 6.0, 6.8 after dissolution in 0.5ml of water.

In some embodiments, the pH adjuster may also act as a permeation enhancer.

In some embodiments, the pharmaceutical composition may comprise epinephrine, an epinephrine prodrug, or a combination of both. The combination of the two includes various combinations of epinephrine and prodrug, and in some embodiments, the compositions provide totally new pharmacokinetic profiles of epinephrine throughout the systemic circulation, providing rapid onset and prolonged onset of therapeutic effects of allergic reactions.

In some embodiments, the polymeric film-forming matrix may comprise a polymer. In some embodiments, the polymer may comprise a water-soluble polymer.

In some embodiments, the polymer may be polyvinyl alcohol. In some embodiments, the polyvinyl alcohol can have a molecular weight of about 27000 daltons, about 31000 daltons, about 47000 daltons, about 67000 daltons, about 125000 daltons.

In some embodiments, the polymer may be a cellulose polymer, in some embodiments, the cellulose polymer may be hydroxypropyl methylcellulose, hydroxypropyl cellulose. In some embodiments, the viscosity of the 2% aqueous solution of hydroxypropyl methylcellulose may be 5mpa.s, 6mpa.s, 15mpa.s, 30mpa.s. In some embodiments, the hydroxypropyl methyl microscopic hydroxypropyl methyl content may be 7.0-12.0%. In some embodiments, the cellulose

In some embodiments, the polymer may be a modified starch, in some embodiments, the modified starch may be a rogue productAnd/>

In some embodiments, the polymer may be polyvinylpyrrolidone, a vinylpyrrolidone/vinyl acetate (6:4) copolymer, or a combination of both.

In some embodiments, the polymer may be a polyvinyl alcohol-polyethylene glycol graft copolymer.

In some embodiments, the pharmaceutical composition may further comprise a stabilizer. Stabilizers may include antioxidants, chelating agents, pH adjusters. In some embodiments, the stabilizer may be 2, 6-di-tert-butyl-p-cresol, sodium metabisulfite, disodium edetate, ascorbic acid, citric acid.

In some embodiments, the pharmaceutical composition may comprise epinephrine, an epinephrine prodrug, and a combination of both. In some embodiments, the adrenergic prodrug can be dipivefrin.

In some embodiments, the epinephrine prodrug may be an ester of epinephrine.

In some embodiments, the pharmaceutical composition may comprise a molecular structure comprising a combination of the following adrenaline and an epinephrine prodrug:

compared with the prior art, the invention has the following beneficial effects:

1. The composition containing epinephrine and the prodrug thereof provided by the invention can quickly increase the epinephrine concentration to a proper effective treatment concentration through a permeation enhancement technology, and maintain the concentration for a period of time, so that adverse reactions caused by the ultra-high peak concentration are avoided, and more convenient and effective treatment selection is brought to allergic patients.

2. The invention researches the pharmacokinetic characteristics of the pharmaceutical composition, verifies the permeability of the medicine through an in vitro permeation test (an in vitro sublingual mucosa permeation test), and verifies the pharmacokinetics of the medicine through a pharmacokinetics test of a beagle dog. The results show that: the composition containing epinephrine and the prodrug thereof provided by the invention has the advantages that the lag time is shortened, permeation occurs more quickly, the exposure time of the drug is prolonged, the concentration fluctuation of the drug in blood plasma can be reduced, adverse reactions caused by the ultra-high peak concentration are reduced, and the treatment effect is prolonged.

Drawings

FIG. 1 pharmacokinetic profile of epinephrine in the prior art (Drebog et al, ALLERGY ASTHMA CLIN Immunol, 2021)

FIG. 2 pharmacokinetic profile AQST-109 of prior art (Aquestive publication, 2021)

FIG. 3 example 1 PK profile prediction of epinephrine and prodrug compositions and epinephrine alone and prodrug alone

FIG. 4 example 4 influence of pH 4 on epinephrine permeability

FIG. 5 effect of pH 4 on dipivefrine

FIG. 6 example 5 effect of donor concentration on the permeability to dipivefrin

FIG. 7 example 11 permeability of adrenergic membranes

FIG. 8 permeability of the dipivefrin film of example 11

FIG. 9 example 14 experiment of the conversion of dipivefrin in plasma

FIG. 10 example 15 ratio of dipivefrin to epinephrine conversion in permeability experiments

FIG. 11 example 15 conversion ratio of dibenzoylmethepinephrine in permeability experiments

Detailed Description

It is to be noted that the raw materials used in the present invention are all common commercial products, and the sources thereof are not particularly limited.

Example 1

This example illustrates predicted pharmacokinetic profiles of epinephrine after combination of epinephrine and dipivefrin. The pharmacokinetic calculation model for the epinephrine and prodrug combination is shown below:

a (1) is a component of the first stage and may be epinephrine or a rapidly converting prodrug. A (2) is a component of the second stage, allowing for slow absorption and conversion of the prodrug.

K _a1 is the first order rate constant following sublingual or nasal administration, K _a2 is the first order rate constant for the overall slow onset of epinephrine in the blood circulation, which may be the slow absorption of prodrug, the slow conversion of prodrug to epinephrine, or a combination of prodrug absorption conversions.

A (3) is the amount of epinephrine in the circulation. Epinephrine is calculated by A (3)/Vd, where Vd is the volume of epinephrine distributed, K _elim is the first order rate constant to eliminate epinephrine from the circulation, calculated by CL/Vd, where CL is the systemic clearance of epinephrine. It is estimated that epinephrine has a CL of about 2L/h/kg and Vd is about 0.08L/kg (Oualha et al, CRITICAL CARE 2014, 18:R23). K _elim is about 25/hr. In some embodiments, additional delays in absorption or conversion may occur, where additional T _lag parameters may be used for pharmacokinetic modeling.

In the pharmacokinetic calculations, it was assumed that the actual total amount of epinephrine and dipivefrin absorbed was an epinephrine dose corresponding to 0.75 μg/kg. This example is used to show the difference in time profile after combination of epinephrine and dipivefrin in different ratios. The actual amount of drug added to the sublingual film or nasal spray or intramuscular injection depends on the actual bioavailability of the different formulations by different routes. The sublingual absorption of epinephrine is rapid (example 3) because epinephrine appears rapidly in the recipient's pool. For such a fast recovery, it can be assumed that K _a1 is a large value, for example 10 1/hour. In vitro permeation studies with porcine sublingual mucosa showed that epinephrine was present in the recipient pool after about 15 minutes and that this uptake was stable within about 2 hours. Thus, total absorption and/or conversion T1/2 is estimated to be 0.4 hours (assuming stabilization after five half-lives), and K _a2 is about 1.5 1/hour.

Formulations of epinephrine, an epinephrine prodrug, or a combination of both, result in differences in pharmacokinetic profiles due to differences in raw materials. Formulation 1 contained 100% epinephrine, dipivefrin; formulation 2 contained 30% epinephrine and 70% dipivefrin (molar ratio), and formulation 3 contained 100% dipivefrin, with no epinephrine. Formulation 1 showed a rapid rise in epinephrine concentration and, to very high values, rapidly dropped to very low levels about 30 minutes. High concentrations of epinephrine may present a higher risk of side effects such as cardiotoxicity. While low concentrations after about 30 minutes have no therapeutic effect, which may increase the likelihood of second-stage allergic reactions occurring, resulting in the need to prepare two doses of the drug. Formulation 3 contained only the prodrug dipivefrin, whose PK profile showed that epinephrine began to appear and climb after a delay of 15-20 minutes. Formulation 2 contained 30% epinephrine and 70% dipivefrin, whose PK profile showed that 1) epinephrine was rapidly marketed in 5-10 minutes; 2) Bimodal and more prolonged drug exposure occurred, with drug exposure times exceeding 1h (see figure 3). Overall, formulation 2, as a combined formulation of the appropriate epinephrine and its prodrug dipivefrin, produced a better therapeutic effect than formulations 1 and 3.

Example 2

In vitro sublingual mucosal permeation protocol

Example in vitro sublingual mucosal penetration is assessed as follows.

(1) The porcine sublingual tissue was freshly resected and rapidly dissected and treated no more than 8 hours prior to treatment with tissue.

(2) Muscles, tissues and the like connected to the sublingual mucosa are removed and cut to a precise thickness.

(3) The mucosa is cut to size and placed in a Franz diffusion cell that includes a donor drug, a donor cell, a mucosal tissue, a recipient cell, a stirrer, and a heater.

(4) 12Ml of receptor Chi Jiezhi (PBS 6.8 buffer) was added to the receptor pool. The donor pool, mucosal tissue were fixed by ligation, ensuring no air bubbles in the recipient pool, and pre-heated at 37 ℃ for half an hour. After the end of the warming up, 0.5mL of donor drug solution or 1cm diameter round membranes were added to the donor pool and wetted with 500uL of PBS 6.8 buffer.

(5) At given time intervals, typically 0.25, 0.5, 1,2, 3 and 6 hours. 400 μl was sampled from the recipient cell by syringe and replenished to the scale with fresh medium, with high care to avoid introducing air into the recipient cell during sampling and replenishing, and if air is introduced, it would need to be immediately drained.

(6) And detecting the concentration of the medicine in the sample by adopting a High Performance Liquid Chromatograph (HPLC). The cumulative amount of drug (mg) per unit surface area penetrated by permeation (cm ²) is plotted against time (h).

Example 3

In vitro permeation experiments, the permeation of epinephrine tartrate under the action of various permeation enhancers, the prodrug dipivefrin hydrochloride under the action of various permeation enhancers, and the prodrug dibenzoylmethane epinephrine (DBE) under the action of various permeation enhancers was studied.

According to the Fick's first law, a steady-state flow Jss, an enhancement factor EF and a permeability coefficient P of the drug in the stratum corneum are calculated by using a following calculation formula.

Equation 1: dM/Sdt=Jss=DC ₀ K/h

Equation 2: p=kd/h

Equation 3: jss=pc ₀

Equation 4: ef=j ₁/J₀

Where D is the coefficient of dispersion of the drug in the stratum corneum, h is the diffusion path length or mucosal thickness, K is the coefficient of distribution of the drug in the stratum corneum and the carrier, C ₀ is the donor concentration, and P is the coefficient of penetration in the stratum corneum. Jss is the amount of drug permeation in the steady-state portion and the slope of the time curve, and Lag time Lag time is the extrapolation of the steady-state linear equation. J0 and J1 represent steady state flow without and with a permeation enhancer, respectively.

Permeation enhancer-epinephrine

Permeation experiments using 4.4mg/ml donor concentrations of epinephrine tartrate and permeation enhancers showed significant permeation enhancement to epinephrine for 3wt% clove oil, 3wt% labrasol, 3wt% angelica oil, 10wt% labrafil, 3% peppermint oil. And except 3wt% labrasol, each group of permeation enhancers shortens the lag time, indicating that permeation increases while steady state flow is achieved at an earlier time and permeation occurs more rapidly. It is explained that clove oil, labrasol, angelica oil, labrafil and peppermint oil have a promoting and enhancing effect on transmucosal permeation of epinephrine. It is expected that similar permeation enhancement results can be obtained for pharmaceutical compounds that are structurally similar to epinephrine.

Table 1 example 3 osmotic data in an epinephrine ex vivo osmotic experiment

Penetration enhancer dipivefrine

In vitro permeation experiments using dipivefrin hydrochloride at a concentration of 8mg/ml and various permeation enhancers showed significant permeation promotion of dipivefrin to Labrasol, labrafil, peppermint oil, levulinic acid and dimethyl sulfoxide (DMSO). And each group of permeation enhancers shortens the delay time, which indicates that the steady-state flow reaches at an earlier time and permeation occurs more quickly while permeation is increased. It is expected that similar permeation enhancement results can be obtained for pharmaceutical compounds that are structurally similar to epinephrine.

Table 2 example 3 osmotic data in the dipivefrin in vitro osmotic experiments

Permeation enhancer dibenzoylmethepinephrine

In vitro permeation experiments using 8mg/ml concentration of dibenzoylmethepinephrine and various permeation enhancers showed that the permeation enhancement of several permeation enhancers was not evident, but Labrasol, transcuto P, peceol all reduced lag time, accelerated permeation time.

TABLE 3 example 3 osmotic data in dibenzoylmethepinephrine ex vivo permeation experiments

Example 4

Influence of pH

In an in vitro sublingual mucosa permeability experiment, the effect of the pH value of the donor drug solution on the permeability was studied, see FIGS. 4-5. In this example, the result of permeability shows that an increase in acidity increases the permeability of epinephrine, especially when the pH is around 1.5. In contrast, pH has little effect on the permeability of dipivefrin and slightly alkaline conditions (e.g., pH 8.4) are beneficial to the permeability of dipivefrin.

Example 5

Influence of drug-carrying concentration

In an in vitro sublingual mucosa permeability experiment, the effect of drug concentration of the donor drug dipivefrin on permeability was studied, see fig. 6. In this example, the results of the permeability show that an increase in concentration increases the permeability of dipivefrin.

Example 6

Study of film-forming materials of sublingual film agents

In experiments where epinephrine and prodrugs were prepared as sublingual films, film forming studies were conducted on different rapidly soluble polymers. The film forming material is dissolved in water, and after the raw materials, the penetration enhancer and other auxiliary materials are added, the film forming material is coated on a release film according to a certain thickness, and after drying and cutting, the sublingual film is obtained. And (5) examining the folding strength, tensile strength and dissolution time of the film agent, and evaluating the film forming property and the drug release speed. In some embodiments, examining polyvinyl alcohol 4-88 as a film forming material, the results show that when polyvinyl alcohol 4-88 is used as a film forming material, the degree of folding resistance can be greater than 30 times, the tensile strength is greater than or equal to 4σM, and the film agent drug is completely released within 2 minutes.

TABLE 4 example 6 evaluation of polyvinyl alcohols 4-88 as film forming materials

Example 7

In one embodiment, the epinephrine/dipivefrin pharmaceutical combination film may be prepared using the following formulation:

TABLE 5 example 7 formulation

Names of raw and auxiliary materials	Wet weight, g	Dry weight, g	Dry weight ratio
				Water and its preparation method	28	0	0％
Polyvinylpyrrolidone	1.5	1.5	11.3％
				Polyvinyl alcohol	7	7	52.6％
Dipivofilin hydrochloride	3	3	22.56％
				Sucralose	0.3	0.3	2.26％
Polyethylene glycol 400	1.5	1.5	11.3％

Example 8

TABLE 6 example 8 formulation

Names of raw and auxiliary materials	Wet weight, g	Dry weight, g	Dry weight ratio
				Water and its preparation method	28	0	0％
Polyvinylpyrrolidone	1.5	1.5	12.7％
				Polyvinyl alcohol	6	6	50.8％
Dipivofilin hydrochloride	1.5	1.5	12.71％
				Epinephrine system	0.5	0.5	4.23％
Sucralose	0.3	0.3	2.54％
				Labrafil	1	1	8.47％
Polyethylene glycol 400	1	1	8.5％

Example 9

TABLE 7 example 9 formulation

Names of raw and auxiliary materials	Wet weight, g	Dry weight, g	Dry weight ratio
				Water and its preparation method	10	0	0％
Propylene glycol	2	2	10.5％
				Polyethylene glycol 400	2	2	10.5％
Ethanol	23	0	0％
				Hydroxypropyl methylcellulose	10.5	10.5	54.9％
Citric acid	0.25	0.25	1.3％
				Sodium metabisulfite	0.38	0.38	2.0％
Tartaric acid epinephrine	1.2	1.2	6.27％
				Dipivofilin hydrochloride	2.8	2.8	14.63％

Example 10

TABLE 8 example 10 formulation

Names of raw and auxiliary materials	Wet weight, g	Dry weight, g	Dry weight ratio
				Water and its preparation method	35	0	0％
Hydroxypropyl methylcellulose	2	2	12.7％
				Propylene glycol	1.5	1.5	9.5％
Polyvinyl alcohol	7	7	44.3％
				Dipivofilin hydrochloride	2.1	2.1	13.3％
Sucralose	0.3	0.3	1.9％
				Polyethylene glycol 400	1.5	1.5	9.5％
Citric acid	0.2	0.2	13％
				Sodium metabisulfite	0.3	0.3	1.9％
Tartaric acid epinephrine	0.9	0.9	5.7％

Example 11

Permeability of sublingual film agent

In an in-vitro sublingual mucosa permeability experiment, the prepared sublingual film agent is subjected to permeability evaluation, and the permeability of the film-formed medicine is inspected. In some embodiments, the film is cut to a diameter of 3/8 inch (0.9525 cm) and placed in a donor cell of a Franz diffusion cell as the donor drug. The drug permeability was examined according to the in vitro sublingual mucosa permeability test procedure, see figures 7-8 and table 9. In some embodiments, the steady flow rate of epinephrine or the prodrug dipivefrin can be up to 0.15 mg/(cm ₂ h) or more and the amount of drug delivered over 0.5 hours can be up to 0.2mg or more. Wherein steady state flow rate of 008-190329C is maximum, and the amount of drug permeated is maximum, and 0.375mg of drug permeated is reached after 0.5 hours.

TABLE 9 permeability data for example 11 films

Example 12

Effect of pH on stability of aqueous solutions of dipivefrin

Dipivefrin was dissolved in water at a concentration of 40-50 μg/mL, pH adjusted to different values with formic acid, phosphoric acid and sodium hydroxide, stored in a constant temperature and humidity cabinet at 25 ℃ (60% rh), and the effect of pH on dipivefrin stability was examined by measuring the concentration of dipivefrin on days 0, 1, 2, 3, 7, 37 and 50. The stability results show that dipivefrin is relatively stable under acidic conditions and degrades over time under slightly alkaline conditions. In some embodiments, dipivefrin remains stable at pH 2.89, pH 3.99, pH 5.27, pH6.36, pH 7.42, with a concentration change of less than 1%. In some embodiments, dipivefrin degrades after 37 days at pH 8.38, and pH 6.75 by an amount greater than or equal to 10%.

TABLE 10 influence of example 12pH on the stability of dipivefrin

Example 13

The stability of the dipivefrin film at both 25 ℃ and 40 ℃ was investigated. In some embodiments, the amount of dipivefrin film remains relatively stable for less than one month under 25 ° conditions, with a fluctuation range of no more than 1%. However, the degradation speed is obviously accelerated at 40 ℃, and the content can be reduced by 7.8% at most after one month, which indicates that the temperature is a key factor affecting the stability of the dipivefrin.

TABLE 11 stability data for the dipivefrin film formulation of example 13

Example 14

Conversion of prodrugs to epinephrine in plasma

The rate and amount of conversion of the prodrug dipivefrin to epinephrine in plasma was studied. In some examples, after pre-heating the heparin-anticoagulated human plasma in an incubator at 37 ℃ for 20 minutes, dipivefrin is added to the plasma at higher concentrations, e.g., 40 μm and 80 μm, in duplicate, at each concentration, 400 μl of plasma is taken at 2,4, 8, 20, 40, 60, and 120 minutes, 160ml of trifluoroacetic acid precipitated protein is added, mixed well, sonicated, the supernatant is filtered, and the prodrug and epinephrine concentration is detected by HPLC injection, as shown in fig. 9. The results show that the prodrug can be metabolized in human plasma and converted to epinephrine. Conversion to epinephrine was detectable from 4-20 minutes, at a concentration of 40. Mu.M at 20 minutes and at a concentration of 80. Mu.M at 4 minutes. From the time of epinephrine appearance to 120min, epinephrine was found to be continuously and slowly rising, indicating that conversion was continuously proceeding. It is predicted that upon administration of the combination formulation of local phenylephrine and epinephrine to an animal or human, in an initial phase, rapid ramp-up of epinephrine will occur in the plasma with osmotic uptake of epinephrine and uptake and conversion of epinephrine, and in a later phase with elimination of epinephrine, but the cumulative sustained conversion of phenylephrine to epinephrine, the plasma may develop a second peak epinephrine concentration or a more prolonged epinephrine exposure profile.

Example 15

In vitro transmucosal permeability experiments, the conversion of prodrugs to epinephrine

In an ex vivo transmucosal permeability experiment, the conversion of prodrug to epinephrine was also detected in the receptor pool. This may be due to the presence of hydrolytic enzymes in the mucosa, the prodrug being hydrolysed by the passage of the prodrug through the mucosa and some of the hydrolytic enzymes falling into the recipient pool. In some embodiments, the results of the permeability experiments of dipivefrin (see fig. 10) show that after 15 minutes epinephrine begins to appear in the recipient pool and plateau is reached after approximately 2 hours, about 70% of the dipivefrin that permeates the mucosa is converted to epinephrine. The plateau phase is assumed to be reached after 5 half-lives, so the half-life T1/2 of the in vitro mucosal to prodrug conversion is 0.4 hours.

In another embodiment, the converted epinephrine is also detected in the ex vivo transmucosal permeation of the prodrug dibenzoylmethepinephrine (see FIG. 11). If only the proportion of the total amount of "substrate + adrenal" is seen, the proportion does not continuously rise as in the dipivefrin permeability experiment described above, but gradually drops after fluctuation. In fact, the study also detected two unknown metabolites, 1 and 2.1 was detected early and peak area was large, decreasing rapidly at 1 hour. 2 is detected only 1 hour and rises rapidly. It is presumed that the metabolite 1 may be a product of hydrolysis of one benzoic acid side chain, and 2 may be side chain benzoic acid separated out upon hydrolysis. In summary, in vitro transmucosal permeability experiments have found that hydrolytic enzymes are also present in the mucosa which hydrolyse the prodrug of epinephrine, and that part of the prodrug can be hydrolyzed to epinephrine during permeation. Thus, it is speculated that during actual administration, a similar phenomenon will occur, where the prodrug is hydrolyzed to epinephrine during permeation, and thus epinephrine may be rapidly present in the plasma, providing the concentration required for treatment.

Example 16

Epinephrine + dipivefrin nasal spray formulation

TABLE 12 example 16 formulation

Names of raw and auxiliary materials	Prescription quantity
		Sodium chloride	0.8g
Epinephrine system	0.9g
		Dipivofilin hydrochloride	4.4g
Sodium metabisulfite	0.1g
		Ethylene diamine tetraacetic acid disodium salt	0.15g
SNAC	10g
		Hydrochloric acid	Proper amount of pH value is regulated to 3-5
20% Ethanol-Water	Constant volume to 100mL

Note that: SNAC is sodium 8- (2-LingYlbenzamido) octoate

Example 17

Epinephrine + dipivefrin nasal spray formulation

TABLE 13 example 17 formulation

Names of raw and auxiliary materials	Prescription quantity
		Sodium chloride	0.8g
Epinephrine system	0.9g
		Dipivofilin hydrochloride	4.4g
Sodium metabisulfite	0.1g
		Ethylene diamine tetraacetic acid disodium salt	0.15g
Capric acid sodium salt	10g
		Hydrochloric acid	Proper amount of pH value is regulated to 3-5
20% Ethanol-Water	Constant volume to 100mL

Example 18

Epinephrine + dipivefrin nasal spray formulation

TABLE 14 example 18 formulation

Names of raw and auxiliary materials	Prescription quantity
		Sodium chloride	0.8g
Epinephrine system	0.9g
		Dipivofilin hydrochloride	4.4g
Sodium metabisulfite	0.1g
		Ethylene diamine tetraacetic acid disodium salt	0.15g
Dodecyl-beta-D-maltoside	3g
		Hydrochloric acid	Proper amount of pH value is regulated to 3-5
20% Ethanol-Water	Constant volume to 100mL

Example 19

Beagle pharmacokinetic experiments with epinephrine + dipivefrin combination formulations

Pharmacokinetic experiments with sublingual film formulations: the beagle dogs are adopted as model animals for animal experiments, 0.5ml of blood is taken at 2h,1h,0.5h,0.25h and 0h before experimental administration, the blood is added into an anticoagulation tube, centrifugation is carried out at 3500rpm for 10min, supernatant plasma is taken, and 0.5M EDTA-Na2 and 6mg/ml sodium metabisulfite are added as stabilizers, so that the stability of epinephrine in the plasma is ensured. And then taking the sublingual film agent of the epinephrine and dipivefrin combination, placing the beagle on a blood drawing frame for fixing, breaking off the mouth of the beagle, placing the sublingual film at a position below the tongue of the beagle, immediately closing the mouth of the beagle, and fixing the mouth of the beagle for more than 15 minutes to avoid swallowing the sublingual film agent. Blood was collected as described above at 3,5, 10, 15, 20, 30, 45, 60, 90 and 120min after administration. After plasma collection, the samples were stored in a refrigerator at-60℃until detection. The concentration of epinephrine and dipivefrin in plasma was detected using liquid chromatography-mass spectrometry (LC/MS).

Pharmacokinetic experiments of nasal sprays: the beagle dogs are adopted as model animals for animal experiments, 0.5ml of blood is taken at 2h,1h,0.5h,0.25h and 0h before experimental administration, the blood is added into an anticoagulation tube, centrifugation is carried out at 3500rpm for 10min, supernatant plasma is taken, and 0.5M EDTA-Na2 and 6mg/ml sodium metabisulfite are added as stabilizers, so that the stability of epinephrine in the plasma is ensured. And then taking the combined nasal spray of epinephrine and dipivefrin, placing the beagle on a blood drawing frame for fixing, inserting a nozzle of the nasal spray into one nostril of the beagle, pressing a spray pump, spraying out a liquid medicine, and carrying out nasal administration. Blood was collected as described above at 3,5,10,15,20,30,45,60,90 and 120min post administration. After plasma collection, the samples were stored in a refrigerator at-60℃until detection. The concentration of epinephrine and dipivefrin in plasma was detected using liquid chromatography-mass spectrometry (LC/MS).

Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.

Claims

1.A composition comprising epinephrine and a prodrug thereof, wherein the pharmaceutically active ingredient comprises a combination of epinephrine and a prodrug of epinephrine.

2. The composition of claim 1, wherein the adrenergic prodrug is a compound of formula (1) or a pharmaceutically acceptable salt thereof:

Wherein R _1a、R_1b、R₂、R₃, independently of one another, can be hydrogen atom H, C-C16 alkyl, alkylcarbamoyl, alkyloxycarbonyl, benzoylmethyl, sulfate or phosphate.

3. The composition of claim 2 wherein when one of the groups R _1a、R_1b、R₂、R₃ is other than H, two of the four groups form a cyclic structure; wherein the structural formula of the epinephrine prodrug comprises dicarbonyl, disulfate or diphosphate.

4. A composition according to claim 3 wherein a cyclic structure is formed by R _1a and R _1b、R_1b and R ₂、R_1b and R ₃、R_1b and R ₂、R_1b and R ₃, or R ₂ and R ₃.

5. The composition of claim 1, further comprising a permeation enhancer.

6. The composition of claim 1, wherein the composition is for sublingual mucosal administration as a film, nasal spray and intramuscular injection.

7. The composition of claim 1, wherein the mass ratio of epinephrine and epinephrine prodrug is between 0.1 and 10.

8. The composition of claim 5, wherein the penetration enhancer comprises glycerides, plant essential oils, fatty acids, esters of oleic acid, diethylene glycol methyl ethyl ether, dimethyl sulfoxide, sodium 8- (2-hydroxybenzoyl) octoate, dodecyl-beta-D-maltoside, and octyl-beta-D-glucopyranoside.

9. The composition of claim 8, wherein the glyceride comprises caprylic capric polyethylene glycol glyceride, oleoyl polyoxyethylene glyceride, or glycerol monooleate.

10. The composition of claim 8, wherein the plant essential oil comprises clove oil, angelica oil, peppermint oil.

11. The composition of claim 8, wherein the fatty acid comprises oleic acid, oleic acid esters, levulinic acid, oleyl oleate, glycerol monooleate, glycerol trioleate.

12. The composition of claim 1, further comprising a stabilizer comprising sodium metabisulfite, ascorbic acid, 2, 6-di-t-butyl-p-cresol, disodium edetate.

13. The composition of claim 1, further comprising a pH adjuster comprising citric acid, hydrochloric acid.

14. The composition of claim 1, further comprising a polymer matrix selected from one or more of hydroxypropyl methylcellulose, polyvinyl alcohol, polyoxyethylene, polyvinylpyrrolidone, modified starch, vinylpyrrolidone/vinyl acetate copolymer, polyvinyl alcohol-polyethylene glycol graft copolymer.

15. Use of a composition according to any one of claims 1 to 14 in a medicament for the treatment of type I allergic reactions.

16. The use according to claim 15, wherein the epinephrine concentration of 100-2000ng/mL is provided within 5 minutes to 6 hours after administration.

17. The use according to claim 16, wherein following administration, an epinephrine concentration of 100-2000ng/mL is provided within 5 minutes to 2 hours.

18. The use of claim 17, wherein the epinephrine concentration of 100-2000ng/mL is provided within 5 minutes to 1 hour after administration.

19. The composition of claim 1, wherein the pharmaceutical composition provides less fluctuation in plasma drug concentration over time than plasma concentration of epinephrine or prodrug alone.

20. The composition of claim 1, wherein the pharmaceutical composition provides: (a) reducing plasma concentration fluctuations; and (b) reducing adverse reactions; (C) prolonged plasma exposure and therapeutic effect.