WO2023167488A1 - Veterinary composition for topical administration of molnupiravir and use thereof - Google Patents

Abstract

A veterinary composition for topical administration of molnupiravir and use thereof are disclosed. The veterinary composition for topical administration according to the present invention is percutaneously absorbed by at least 60% of molnupiravir within 20 minutes of application to the skin and at least 95% of molnupiravir within 60 minutes thereof, thereby significantly enhancing dermal penetration so that the side effects of injection or oral administration can be minimized while comparable efficacy to that of injection or oral administration is obtained.

Description

Veterinary composition for topical administration of molnupiravir and its use

The present invention relates to a veterinary composition for topical administration of molnupiravir and its use, and more particularly, to a veterinary composition for topical administration of molnupiravir having improved skin permeation by having a percutaneous absorption rate of 95% or more within 60 minutes of topical administration. Compositions and their use in the treatment of feline infectious peritonitis.

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

Molnupiravir is an oral antiviral drug developed for the treatment of influenza. As a prodrug of the synthetic nucleoside derivative N4-hydroxycytidine, it 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 for the treatment of COVID-19 as an oral drug.

Recently, molnupiravir has also been reported to have a therapeutic effect on Feline Infectious Peritonitis (FIP) (Non-Patent Document 1).

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

FIP is a progressive immune-related disease, which can take the form of 'wet' or 'dry' FIP. In wet type FIP, blood leaks out of blood vessels weakened by inflammation, causing ascites and pleural effusion, resulting in water filling not only the abdomen but also the lungs, making breathing difficult. In dry type FIP, lesions appear in the stomach, intestines, liver, kidneys, and nervous system, as well as loss of appetite, fever, jaundice, diarrhea, weight loss, and seizures. do it too The onset of FIP, either wet or dry, is invariably fatal. FIP is the leading 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 therapeutics is an antiviral drug targeting a specific protein essential for viral replication (FIPV). GC376, a protease inhibitor of FIPV, GS-441524 of the active part, a precursor of remdesirvir, and remdesirvir (GS-441524) 5734) was reported from 2016 to 2019, and molnupiravir is reported to be similar to remdesivir (Non-Patent Document 1).

However, conventional FIP treatment has been attempted to be developed as an injection or oral drug, and when administered through injections (usually 30 to 100 times) of FIP patients, problems such as soreness, pain, scarring at the injection site, ulcers, scabs, necrosis, etc. Oral administration also requires difficulties and efforts, and side effects such as stress, gastrointestinal disorder, anorexia, and vomiting occur in the sick cat.

Therefore, there is a need for the development of a formulation capable of administering a FIP treatment through topical transdermal without the above problems or side effects, and in particular, the development of a composition for topical administration with high skin permeability in a short time is required.

(Patent Document 1) WO2018/169946

(Non-Patent Document 1) 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, a veterinary composition for topical administration of molnupiravir using a combination of a solvent and a solubilizing agent of a specific composition showed that molnupiravir was 20 The present invention was completed after confirming that skin permeation was remarkably improved by percutaneous absorption of 60% or more within minutes and 95% or more within 60 minutes.

Accordingly, an object of the present invention is to provide a veterinary composition for topical administration of molnupiravir with enhanced skin penetration.

Another object of the present invention is to provide a method for 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 enhanced skin permeation 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), hexamethylphosphoramide In the group consisting of HMPA), hexamethylphosphoric triamide, nitromethane, propylene carbonate, dimethylpropyleneurea, pyridine and sulfolane It is 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, preferably diethylene glycol monoethyl ether.

In the present invention, the weight ratio of the solvent and the solubilizing agent is 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 the solubilizing agent are used in combination in the above weight ratio, the veterinary composition for topical administration of the present invention has a percutaneous absorption of 60% or more of molnupiravir within 20 minutes and 95% or more of molnupiravir within 60 minutes. skin permeation was remarkably improved (Example 1). When the mixed weight ratio of the solvent and the solubilizing agent was out of the above range, the percutaneous absorption rate was lowered (Comparative Examples 1 to 3).

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

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

The pour-on formulation refers to a liquid formulation that can be poured and applied to a portion of the skin along the dorsal line of a 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, in addition, antioxidants, anti-irritation agents, dyes, preservatives, stabilizers, buffers, lubricants, diluents, crystallization inhibitors, thickeners, solubilizers, fluidizing agents, which can be commonly used in the pharmaceutical field at the time of formulation , 0.01 to 10% by weight, preferably 0.05 to 7% by weight of one or more additives selected from the group consisting of a complexing agent, a mineral, an antifoaming agent and a spreading agent.

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. Preferred is butylated hydroxytoluene.

The anti-irritation agent may be selected from pharmaceutically or veterinarily available anti-irritation agents. Preferably it is Povidone K30.

The preservative may be selected from methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, phenol, sorbic acid, cresols and chlorocresols, 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 92 to 92% by weight of a mixture of DMSO and diethylene glycol monoethyl ether. 94.03% by weight, and DMSO and diethylene glycol monoethyl ether are mixed at 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 preparing a composition for topical administration. That is, the composition may be prepared by completely dissolving molnupiravir in a solvent and a solubilizing agent by a mechanical method, and then mixing and dissolving additives. In addition, the composition is a solution in which molnupiravir is first dissolved in a part of the solvent, and the rest It may be prepared 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, 10 to 50 mg/ml of molnupiravir, 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 cat's skin from the head to the tail.

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

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

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

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

In the present invention, 'feline subject' means a cat in need of 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, 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, preferably topically administered (applied) in a volume of 1 ml or less.

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

The veterinary composition for topical administration according to the present invention is percutaneously absorbed by more than 60% of molnupiravir within 20 minutes and more than 95% of molnupiravir within 60 minutes of application to the skin, thereby significantly enhancing skin permeation, It is possible to minimize the side effects of injection or oral administration while achieving the corresponding efficacy.

1 is a photograph showing a veterinary composition for topical administration prepared in the present invention. 1: Comparative Example 1, 2: Example 1, 3: Comparative Example 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 percutaneous absorption rate over time of the veterinary composition for topical administration prepared in the present invention.

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

Preparation Example 1: Preparation of veterinary composition for topical administration

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

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

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 permeation (percutaneous 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, the skin permeation (percutaneous absorption rate) test was performed using the Transdermal Diffusion Cell System DHC-6TD model (LOGAN Instruments (USA)). proceeded using

The Transdermal Diffusion Cell System DHC-6TD model is a device capable of completely removing air bubbles from media by using a Bubble Free Franz Cell and a Franz Cell Plate Tilting function. A Franz diffusion cell with an inner diameter of 15 mm and an inner volume of 12 ml (13 ml when added to the connection line volume) was used, 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 so that the test substance did not leak, and the upper part was assembled. The lower part of the Franz diffusion cell was filled with ultrapure water, which is an aqueous solution (receptor fluid). The internal temperature was set to 37 °C and stirred using a magnetic bar. When the temperature reaches the set temperature, 0.5 ml (containing about 14.25 mg of molnupiravir) of each sample (composition of Example 1 and Comparative Examples 1 to 3) was applied on the artificial skin PB-M, and at 20 minutes and 1 hour The aqueous solution was aliquoted. The fractionation method proceeded by discharging the entire aqueous solution and filling it with a new aqueous solution. The fractionated aqueous solution was quantitatively analyzed by HPLC.

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

Specifically, a solution in which 4.6 g of ammonium dihydrogen phosphate was 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 a sample solution (Ammonium dihydrogen phosphate solution: ACN = 95: 5) to prepare a concentration of 3,000 ppm, 1500 ppm, 750 ppm, 300 ppm, 150 ppm, 60 ppm, 30 ppm, 15 ppm, 3 ppm, 1.5 ppm concentration A calibration curve was prepared with the area value obtained by dilution with (FIG. 2). Using the prepared calibration curve, the aqueous solution fractionated for each time period in the percutaneous absorption experiment was quantitatively analyzed to calculate the percutaneous absorption rate, and the results are shown in Table 5 and FIG. 3.

column	YMC-Triart C18 plus / 150*4.6mm / 3um
mobile phase	Ammonium dihydrogen phosphate solution : MeOH
flow rate	1.0ml/min
Detector	DAD (260 nm)
hour	53min
column temperature	40 ℃
infusion capacity	5 μl
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 percutaneous 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 (total)	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) was able to confirm that the percutaneous absorption rate of molnupiravir after skin application was significantly increased to 62.73% within 20 minutes and 97.08% within 60 minutes. there is.

Claims (12)

1. A veterinary composition for topical administration of molnupiravir with enhanced skin permeation comprising molnupiravir, a solvent and a solubilizing agent,

   The solvent and the solubilizing agent are included in a weight ratio of 1: 1.5 to 2.4,

   Molnupiravir is included in 1 to 10% by weight,

   The composition is a composition that has a transdermal absorption rate of 60% or more and 95% or more within 60 minutes within 20 minutes of molnupiravir being applied to the skin.
2. 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 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.
3. The veterinary composition according to claim 2, wherein the solvent is dimethyl sulfoxide and the solubilizing agent is diethylene glycol monoethyl ether for topical administration of molnupiravir with enhanced skin penetration.
4. The method of claim 1, wherein the composition is an antioxidant, anti-irritant, dye, preservative, stabilizer, buffer, lubricant, diluent, crystallization inhibitor, thickener, solubilizer, fluidizing agent, complexing agent, mineral, antifoaming agent and spreading agent. A veterinary composition for topical administration of molnupiravir with enhanced skin permeation, further comprising 0.01 to 10% by weight of one or more additives selected from.
5. The veterinary composition of claim 1, wherein the composition further comprises butylated hydroxytoluene as an antioxidant and povidone K30 as an anti-irritation agent for topical administration of molnupiravir with enhanced skin penetration.
6. The method of claim 1, wherein the composition is molnupiravir 2 ~ 3% by weight, butylated hydroxytoluene 0.07 ~ 0.2% by weight, povidone K30 3 ~ 5% by weight, a mixture of dimethyl sulfoxide and diethylene glycol monoethyl ether 92 to 94.03% by weight, and dimethyl sulfoxide and diethylene glycol monoethyl ether are mixed at a weight ratio of 1: 1.7 to 2.1, a veterinary composition for topical administration of molnupiravir with enhanced skin penetration.
7. The veterinary composition according to claim 1, wherein the composition is a spot-on formulation or a pour-on formulation for topical administration of molnupiravir with enhanced skin penetration.
8. The veterinary composition for topical administration of molnupiravir with enhanced skin permeation according to claim 1, wherein the composition contains 10 to 50 mg/ml of molnupiravir.
9. A method for 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.
10. 10. The method of claim 9, wherein the composition is a spot-on formulation or a pour-on formulation.
11. The method of claim 9, wherein the composition is applied in a volume of 5 ml or less.
12. 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.

Images