KR20230129922A - Veterinary composition for topical delivery of molnupiravir and its uses - Google Patents

Abstract

The present invention discloses a veterinary composition for topical administration of molnupiravir and its use.
In the veterinary composition for topical administration according to the present invention, more than 60% of molnupiravir is absorbed percutaneously within 20 minutes of application to the skin, and more than 95% of molnupiravir is absorbed transdermally within 60 minutes, thereby significantly improving skin penetration, making it effective when administered by injection or oral administration. It is possible to achieve equivalent medicinal efficacy while minimizing side effects during injection or oral administration.

Description

Veterinary composition for topical delivery of molnupiravir and its uses {Veterinary composition for topical delivery of molnupiravir and its uses}

The present invention relates to a veterinary composition for topical administration of molnupiravir and its use. More specifically, it relates to a veterinary composition for topical administration of molnupiravir, which has a percutaneous absorption rate of 95% or more within 60 minutes of topical administration and has improved skin penetration. It relates to a composition and its use in the treatment of feline infectious peritonitis.

Molnupiravir (EIDD-2801) has the following chemical formula: ((2R,3S,4R,5R)-3,4-dihydroxy-5-(4-hydroxyimino)-2-oxo-3 ,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methylisobutyrate [((2R,3S,4R, 5R)-3,4-dihydroxy-5-(4- (hydroxyimino)-2-oxo-3,4-dihydropymidin-1(2H)-yl)tetrahydrofuran-2-yl)methylisobutyrate]:

Molnupiravir is an oral antiviral drug developed for the treatment of influenza. A prodrug of the synthetic nucleoside derivative N4-hydroxycytidine, which exhibits antiviral activity by inducing replication errors during viral RNA replication, resulting in antiviral effects against RNA viruses, including MERS-CoV and SARS-CoV-2. has been reported and has been conditionally approved as an oral drug for the treatment of COVID-19.

Recently, molnupiravir was also reported to be effective in treating feline infectious peritonitis (FIP) (Non-patent Document 1).

Feline coronavirus (FCoV) exists as feline enteric coronavirus (FECV) and a mutant form of FECV, feline infectious peritonitis virus (FIPV). FECV-infected cats are common, are asymptomatic, and do not cause pathological problems. However, FIPV, a mutant form of FECV, causes feline infectious peritonitis (FIP).

FIP is a progressive immune-related disease and can take the form of ‘wet’ or ‘dry’ FIP. Wet type FIP occurs when blood flows out of blood vessels weakened by inflammation, causing ascites and pleural effusion, causing water to fill not only the abdomen but also the lungs, making breathing difficult. Dry type FIP causes lesions in the stomach, intestines, liver, kidneys, and nervous system, as well as decreased appetite, fever, jaundice, diarrhea, weight loss, and seizures. Eye disease and neurological symptoms cause walking problems and paralysis. Sometimes it happens. The onset of FIP, either wet or dry, is invariably fatal. FIP is a major cause of death in cats under 2 years of age, and is estimated to kill 0.3 to 1% of cats worldwide (Patent Document 1).

Research on FIP treatments is an antiviral drug targeting a specific protein (FIPV) essential for viral replication. GC376, a protease extract of FIPV, GS-441524, the active part of the precursor of remdesirvir, and remdesirvir (GS- 5734) was reported from 2016 to 2019, and molnupiravir is reported to be similar to remdesirvir (Non-patent Document 1).

However, attempts are being made to develop conventional treatments for FIP as injections or oral drugs. When administered to cats with FIP through injections (usually 30 to 100 times), problems such as swelling, pain, scarring at the injection site, ulcers, scabs, and necrosis occur. Oral administration is also difficult and requires effort, and side effects such as stress in the cat, gastrointestinal disorders, loss of appetite, and vomiting occur.

Therefore, there is a need for the development of a formulation that can administer FIP treatment through topical transdermal treatment without causing the above-mentioned problems or side effects. In particular, the development of a composition for topical administration with high skin permeability in a short period of time is required.

WO2018/169946

Pedersen N.C. The Long History of Beta-d-n4-Hydroxycytidine and Its Modern Application to Treatment of COVID-19 in People and FIP in Cats. Sock it to Fip. October 20, 2021 [https://sockfip.org/https-sockfip-org-wp-content-uploads-2022-04-the-long-history-of-beta-d-n4-hydroxycytidine-and-its-modern -application-to-treatment-of-covid-19-in-people-and-fip-in-cats-v2-pdf/]

Accordingly, the present inventor continued research to develop a veterinary composition for topical administration of molnupiravir, and as a result, the veterinary composition for topical administration of molnupiravir using a combination of a solvent and a solubilizing agent of a specific composition was found to be molnupiravir 20 The present invention was completed after confirming that skin penetration was significantly improved by transdermal absorption of more than 60% within minutes and more than 95% within 60 minutes.

Therefore, the purpose of the present invention is to provide a veterinary composition for topical administration of molnupiravir with improved skin penetration.

Another object of the present invention is to provide a method of treating feline infectious peritonitis by topically applying the veterinary composition for topical administration of molnupiravir to the skin of a cat.

In order to achieve the above object, the present invention provides a veterinary composition for topical administration of molnupiravir with improved skin penetration, comprising molnupiravir, a solvent, and a solubilizing agent.

In the present invention, the solvent is dimethyl sulfoxide (DMSO), dimethylformamide (DMF), acetone, acetonitrile, ethyl acetate, dichloromethane (DCM), tetrahydrofuran (THF), and hexamethylphosphoramide. ; HMPA), hexamethylphosphoric triamide, nitromethane, propylene carbonate, dimethylpropyleneurea, pyridine, and sulfolane. One or more selected, preferably DMSO.

In the present invention, the solubilizing agent is at least one selected from the group consisting of diethylene glycol monoethyl ether, isopropyl alcohol, methanol, ethanol, propanol, ammonia, formic acid, and acetic acid, and is preferably diethylene glycol monoethyl ether.

In the present invention, the solvent and solubilizing agent are used in a weight ratio of 1:1.5 to 2.4, preferably 1:1.7 to 2.1, and most preferably 1:1.9 to 2.0.

Surprisingly, when the solvent and solubilizing agent are used in combination in the above weight ratio, the veterinary composition for topical administration of the present invention absorbs more than 60% of molnupiravir within 20 minutes and more than 95% of molnupiravir within 60 minutes. Skin penetration was significantly improved (Example 1). When the mixing weight ratio of solvent and solubilizing agent was outside the above range, the transdermal absorption rate decreased (Comparative Examples 1 to 3).

In the present invention, the veterinary composition for topical administration may be a spot-on formulation or a pour-on formulation.

Spot-on formulation refers to a liquid formulation that can be applied dropwise or coated to a portion of the skin from the head to the tail of the target animal.

Pour-on formulation refers to a liquid formulation that can be poured and applied to a portion of the skin along the back line of the target animal.

In the present invention, 'veterinary composition' means a composition suitable for use in the treatment, control, eradication, alleviation or prevention of diseases or symptoms in animals.

In the composition of the present invention, molnupiravir is included at 1 to 10% by weight, preferably at 2 to 9% by weight, and most preferably at 2 to 3% by weight.

The composition of the present invention further contains antioxidants, anti-irritant agents, dyes, preservatives, stabilizers, buffers, lubricants, diluents, crystallization inhibitors, thickeners, solubilizers, and fluidizing agents, which can be commonly used in the pharmaceutical field when formulated. , complexing agents, minerals, antifoaming agents, and spreading agents may be included in an amount of 0.01 to 10% by weight, preferably 0.05 to 7% by weight.

Antioxidants include butylated hydroxytoluene, vitamin E, vitamin E phosphate, vitamin A, ascorbic acid, vitamin B12, polyphenols, butyl hydroxyanisole (BHA), propyl gallate, tocopherol, ascorbic acid, citric acid, di- Alpha-tocopheryl phosphate, beta-carotene, carotene, carotenoids, flavonoids, sulfate compounds, L-cysteine, thiodipropionic acid, thiolactic acid, monothioglycerol, propyl gallate sodium metabisulfite, sodium formaldehyde, sulfoxylate acetate, and mixtures thereof. Preferably it is butylated hydroxytoluene.

The antiirritant agent may be selected from pharmaceutically or veterinarily usable antiirritant agents. Preferably it is povidone K30.

Preservatives may be selected from methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, phenol, sorbic acid, cresol and chlorocresol, and mixtures thereof.

Thickeners include methylcellulose, ethylcellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, and mixtures thereof.

Complexing agents include EDTA and its salts, phosphates, nitrates, acetates, citrates, and mixtures thereof.

Preferably, the composition of the present invention may further include butylated hydroxytoluene as an antioxidant and povidone K30 as an anti-irritant agent.

Preferably, the composition of the present invention contains 2 to 3% by weight of molnupiravir, 0.07 to 0.2% by weight of butylated hydroxytoluene, 3 to 5% by weight of povidone K30, and a mixture of DMSO and diethylene glycol monoethyl ether at 92 to 92%. It contains 94.03% by weight, and DMSO and diethylene glycol monoethyl ether are mixed in a weight ratio of 1:1.7 to 2.1.

The veterinary composition for topical administration of the present invention can be prepared by a conventional method for producing a composition for topical administration. In other words, the composition can be prepared by completely dissolving molnupiravir in a solvent and a solubilizing agent by a mechanical method, and then adding and dissolving additives. Additionally, the composition includes a solution in which molnupiravir is first dissolved in a portion of the solvent, and the remainder. It may also be manufactured by mixing a solution in which an additive is dissolved in a mixture of a solvent and a solubilizing agent.

The veterinary composition for topical administration according to the present invention contains, but is not limited to, molnupiravir in an amount of 10 to 50 mg/ml, preferably 20 to 30 mg/ml.

The veterinary composition for topical administration according to the present invention can be administered by applying it to a part of the skin from the head to the tail of the cat.

The daily dosage for cats of the veterinary composition for topical administration according to the present invention is preferably 1 to 50 mg/kg of molnupiravir, and the single dose is 1 to 25 mg/kg of molnupiravir. It is desirable to set it to kg.

The veterinary composition for topical administration according to the present invention exhibits a transdermal absorption rate of molnupiravir of more than 60% within 20 minutes of application to the skin and more than 95% of molnupiravir within 60 minutes (Test Example 1).

Therefore, the veterinary composition for topical administration according to the present invention has excellent transdermal absorption rate of molnupiravir in a short period of time, and can achieve the corresponding medicinal effect upon injection or oral administration while minimizing side effects during injection or oral administration.

According to another object of the present invention, the present invention provides a method of treating feline infectious peritonitis, comprising topically applying an effective amount of the veterinary composition for topical administration to the skin of a cat.

In the present invention, 'cat individual' refers to a cat that requires treatment for a feline infectious disease.

In the treatment method of the present invention, the veterinary composition for topical administration is a spot-on formulation or a pour-on formulation, and is preferably a spot-on formulation.

In the treatment method of the present invention, the veterinary composition for topical administration is administered in a volume of 5 ml or less, and is preferably administered (applied) topically in a volume of 1 ml or less.

Preferably, it includes topically administering (applying) a veterinary composition for topical administration in a volume capable of delivering 1 to 50 mg of molnupiravir per 1 kg of body weight of the cat.

In the veterinary composition for topical administration according to the present invention, more than 60% of molnupiravir is absorbed percutaneously within 20 minutes of application to the skin, and more than 95% of molnupiravir is absorbed transdermally within 60 minutes, thereby significantly improving skin penetration, making it effective when administered by injection or oral administration. It is possible to achieve equivalent medicinal efficacy while minimizing side effects during injection or oral administration.

1 is a photograph showing a veterinary composition for topical administration prepared in the present invention. 1: Comparative Examples 1, 2: Examples 1, 3: Comparative Examples 2, 4: Comparative Example 3.
Figure 2 is a graph showing the calibration curve of molnupiravir in HPLC quantitative analysis.
Figure 3 is a graph showing the transdermal absorption rate over time of the veterinary composition for topical administration prepared in the present invention.

The invention is explained in more detail by the following examples. These examples are intended to illustrate the invention, and the scope of the invention should not be limited by them.

Preparation Example 1: Preparation of veterinary composition for topical administration

After putting the same volume of DMSO (solvent) and diethylene glycol monoethyl ether (solubilizing agent) in a conical tube as shown in Table 1, molnupiravir was added, followed by vortexing (5 minutes) and ultrasonic treatment (1 minute). After complete dissolution, butylated hydroxytoluene (antioxidant) and povidone K30 (anti-irritation agent) are added, followed by vortexing (5 minutes) and sonication (1 minute) to completely dissolve, thereby preparing a veterinary composition for topical administration. (Figure 1).

Raw material name Comparative Example 1 Example 1 Comparative Example 2 Comparative Example 3 Molnupiravir 285mg 285mg 285mg 285mg Butylated hydroxytoluene 9mg 9mg 10mg 12mg Povidone K30 450mg 473mg 512mg 584mg DMSO 2.469 g 3.462 g 5.666 g 10.919g Diethylene glycol monoethyl ether 7.206 g 6.627 g 5.122 g 1.169 g

The molnupiravir content (mg/ml) in the prepared veterinary composition for topical administration is shown in Table 2.

Content (mg/ml) Comparative Example 1 Example 1 Comparative Example 2 Comparative Example 3 Molnupiravir 28.51 28.51 28.51 28.51

Test Example 1: Skin penetration (transdermal absorption rate) test

For each of the veterinary compositions for topical administration of Example 1 and Comparative Examples 1 to 3 prepared in Preparation Example 1, a skin penetration (percutaneous absorption rate) test was performed using the Transdermal Diffusion Cell System DHC-6TD model (LOGAN Instruments (USA)). It was carried out using

The Transdermal Diffusion Cell System DHC-6TD model is a device that can completely remove air bubbles from media using the Bubble Free Franz Cell and Franz Cell Plate Tilting functions. The Franz diffusion cell was used with an inner diameter of 15 mm and an internal volume of 12 ml (13 ml when including the connecting line volume), and PB-M artificial skin made of cellulose membrane and an inner diameter of 15 mm and an outer diameter of 38 mm were used. , a gasket with a thickness of 1 mm, and a magnetic bar were used.

Specifically, the prepared artificial skin PB-M was placed on a receptor chamber, a gasket was installed to prevent the test substance from leaking, and the upper part was assembled. The lower part of the Franz diffusion cell was filled with ultrapure water, an aqueous solution (receptor fluid). The internal temperature was set at 37°C and stirred using a magnetic bar. When the temperature reaches the set temperature, 0.5 ml of each sample (composition of Example 1 and Comparative Examples 1 to 3) (containing about 14.25 mg of molnupiravir) is applied on the artificial skin PB-M and applied for 20 minutes and 1 hour. The aqueous solution was aliquoted. The preparative method was carried out by discharging the entire aqueous solution and filling it with new aqueous solution. The aqueous solution collected was quantitatively analyzed by HPLC.

HPLC quantitative analysis was performed on each aqueous solution using 1260 high-performance liquid chromatography (HPLC; Agilent) under the conditions shown in Tables 3 and 4 below.

Specifically, a solution of 4.6 g of ammonium dihydrogen phosphate dissolved in 2 L of ultrapure water was analyzed under gradient conditions with mobile phase A and methanol as mobile phase B. First, the standard solution was prepared by dissolving molnupiravir in the sample solution (Ammonium dihydrogen phosphate solution: ACN = 95:5) to prepare a concentration of 3,000ppm, and the concentrations were 1500ppm, 750ppm, 300ppm, 150ppm, 60ppm, 30ppm, 15ppm, 3ppm, and 1.5ppm. A calibration curve was created using the area values obtained by diluting (Figure 2). Using the prepared calibration curve, the aqueous solution collected at each time period in the transdermal absorption experiment was quantitatively analyzed to calculate the percutaneous absorption rate, and the results are shown in Table 5 and Figure 3.

column YMC-Triart C18 plus / 150*4.6mm / 3um mobile phase Ammonium dihydrogen phosphate solution: MeOH flow rate 1.0ml/min Detector DAD(260nm) hour 53min column temperature 40℃ injection dose 5㎕ sample solution Ammonium dihydrogen phosphate solution: ACN = 95:5

hour
(minute) Mobile phase A
(% v/v) Mobile phase B
(% v/v) 0-2 100 0 2-43 100 → 20 0 → 80 43-48 20 80 48-48.1 20 → 100 80 → 0 48.1-53 100 0

Time (minutes) Molnupiravir transdermal absorption rate (%) Comparative Example 1 Example 1 Comparative Example 2 Comparative Example 3 0 - 20 minutes 16.29 62.73 8.99 7.84 20 - 60 minutes 31.86 34.34 36.76 31.17 0 - 60 minutes (cumulative) 48.15 97.08 45.75 39.00

As shown in Table 5, only the veterinary composition for topical administration according to the present invention (Example 1) significantly improved the transdermal absorption rate of molnupiravir to 62.73% within 20 minutes and 97.08% within 60 minutes after skin application. there is.

Claims (12)

A veterinary composition for topical administration of molnupiravir with improved skin penetration, comprising molnupiravir, a solvent, and a solubilizing agent,
Solvent and solubilizing agent are included in a weight ratio of 1:1.5 to 2.4,
Molnupiravir is included in an amount of 1 to 10% by weight,
The composition is a composition in which molnupiravir has a transdermal absorption rate of 60% or more within 20 minutes and 95% or more within 60 minutes of application to the skin.
The method of claim 1, wherein the solvent is dimethyl sulfoxide, dimethylformamide, acetone, acetonitrile, ethyl acetate, dichloromethane, tetrahydrofuran, hexamethylphosphoramide, hexamethylphosphorictriamide, nitromethane, propylene carbonate, At least one selected from the group consisting of dimethylpropylene urea, pyridine, and sulfolane,
The solubilizing agent is a veterinary medicine for topical administration of molnupiravir with improved skin penetration, wherein the solubilizing agent is at least one selected from the group consisting of diethylene glycol monoethyl ether, isopropyl alcohol, methanol, ethanol, propanol, ammonia, formic acid, and acetic acid. Composition.
The veterinary composition for topical administration of molnupiravir with improved skin penetration according to claim 2, wherein the solvent is dimethyl sulfoxide and the solubilizing agent is diethylene glycol monoethyl ether.
The method of claim 1, wherein the composition is a group consisting of antioxidants, anti-irritation agents, dyes, preservatives, stabilizers, buffers, lubricants, diluents, crystallization inhibitors, thickeners, solubilizers, fluidizing agents, complexing agents, minerals, anti-foaming agents, and electrodeposition agents. A veterinary composition for topical administration of molnupiravir with improved skin penetration, further comprising 0.01 to 10% by weight of one or more additives selected from.
The veterinary composition for topical administration of molnupiravir with improved skin penetration according to claim 1, wherein the composition further comprises butylated hydroxytoluene as an antioxidant and povidone K30 as an antiirritant.
The method of claim 1, wherein the composition contains 2 to 3% by weight of molnupiravir, 0.07 to 0.2% by weight of butylated hydroxytoluene, 3 to 5% by weight of povidone K30, and a mixture of dimethyl sulfoxide and diethylene glycol monoethyl ether. A veterinary composition for topical administration of molnupiravir with improved skin penetration, comprising 92 to 94.03% by weight, and dimethyl sulfoxide and diethylene glycol monoethyl ether mixed at a weight ratio of 1:1.7 to 2.1.
The veterinary composition for topical administration of molnupiravir with improved skin penetration according to claim 1, wherein the composition is a spot-on formulation or a pour-on formulation.
The veterinary composition for topical administration of molnupiravir with improved skin penetration according to claim 1, wherein the composition contains molnupiravir at 10 to 50 mg/ml.
A method of treating feline infectious peritonitis comprising topically applying an effective amount of the veterinary composition for topical administration according to any one of claims 1 to 8 to the skin of a cat.
The method of claim 9, wherein the composition is a spot-on formulation or a pour-on formulation.
The method of claim 9, wherein the composition is applied in a volume of 5 ml or less.
The method of claim 9, wherein the composition is applied in a volume capable of delivering 1 to 50 mg of molnupiravir per 1 kg of body weight of the cat.