JPS622565B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to the use of substituted furanaldehydes and alkyl ketones as antirinovirus agents. Compounds of the following formula are useful as antirinoviral agents. In the above formula, R ₂ is a tetradecyl group and R ₃ is a methyl group. In the above formula, the substituent also represented by R2 _- O- may be attached to the 5-position of the furan ring. The present invention relates to the use of compounds of the formula as antirinoviral agents. R ₃ is hydrogen and R ₂ is a straight or branched saturated hydrocarbon chain with 10 to 20 carbon atoms or a straight or branched chain with 10 to 20 carbons and 1 to 4 double bonds Compounds replacing the formula definition when the chain is an unsaturated hydrocarbon chain are disclosed in pending U.S. patent application Ser. The use of this class of compounds as depressants has been described. R ₃ is methyl, R ₂ is a straight or branched saturated hydrocarbon chain with 10 to 20 carbon atoms, or a straight chain with 10 to 20 carbon atoms and 1 to 4 double bonds, or Compounds replacing the formula definitions when they are branched unsaturated hydrocarbon chains are disclosed as intermediates for the production of hypolipidemic agents in pending U.S. Patent Application No. 587,117 filed June 16, 1975. Are listed. R ₃ is methyl and R ₂ is a straight or branched hydrocarbon chain with 10 to 20 carbon atoms or a straight or branched chain with 10 to 20 carbon atoms and 1 to 4 double bonds. Compounds replacing the definition of the formula when the chain is an unsaturated hydrocarbon chain are described as intermediates for the production of hypolipidemic agents in pending U.S. Patent Application No. 347,232, filed April 2, 1973. has been done. Compounds of formula are useful as antirinoviral agents. The genus Rhinovirus, a member of the Picornaviridae family, includes over 100 different serotypes and is known to be responsible for many syndromes associated with respiratory infections. The name rhinovirus refers to the prominent nasal complications seen in these viral infections that produce symptoms characteristic of the common cold. The rhinovirus group includes serotypes 1 to 89 and subtypes 1A (88,
89, 90), and at least 20 more types will be added to the classification. Experimental studies have shown that the nasal mucosa is more susceptible to rhinoviruses than the lower respiratory tract. Symptoms of rhinovirus infection have also been produced experimentally by instillation of small amounts of virus onto the conjunctiva, indicating that the eyeball is another susceptible site for infection. Advanced rhinovirus infections are characterized by hyperemia and edema of the mucous membranes, with exudation of serous fluid and mucus.
The nasal cavity becomes narrow due to thickening of the membranes and hyperemia of the nasal turbinates. The compounds described herein have been found to be effective antiviral agents against many types of rhinoviruses, and have been found to be effective antivirals against rhinovirus infections in susceptible hosts, including humans and some great apes such as chimpanzees. This makes this class of compounds useful for treating conditions. It is known in the art that several test formats can be used to measure antiviral activity against rhinoviruses. For example, anti-rinoviral activity can be measured using plaque assays or in vitro assays, in which the activity of a compound against viral challenge is measured in a cellular context. Using various test systems, compounds of the formula were found to be effective antirinoviral agents when the test compound was given prior to, concurrently with, or after viral challenge. The utility of the compounds described herein as antirinoviral agents has been demonstrated in a variety of test systems. For example, use HeLa cell culture medium and challenge it with 30 to 100 TCID ₅₀ rhinoviruses simultaneously with the test compound at 4, 20 or 100 μl.
g/ml, after which the cell culture medium is incubated for 48 hours. Microscopic examination of cell culture media showed that the compounds of the formula significantly inhibited the cytopathological effects of the virus when compared to cell culture media containing virus challenge. For example, the compound of Example 1 at a concentration of 4 μg/ml
When added to the cell culture medium together with 100 TCID ₅₀ rhinovirus challenge, the cytopathological effects of the virus were inhibited by 87% compared to the control group. In an in vitro assay using HeLa cells, the tissue culture medium ED ₅₀ of the compound of Example 1 was found to be 0.3 μg/ml. For the treatment of rhinovirus infection symptoms, compounds of the formula can be administered orally, topically, e.g. intranasally, and parenterally.
For example, it can be administered intramuscularly. Local administration is preferred. The compounds are preferably administered in the form of a pharmaceutical formulation to a host susceptible to rhinovirus infection, before or after viral invasion, or after the onset of infection. Prophylactic treatment includes approximately 1 to 5 doses prior to anticipated viral exposure.
It is contemplated that an anti-rinoviral effective amount of the compound will be administered for about 5 to 10 days or about 5 to 15 days after exposure to rhinovirus. Rhinoviruses are easily transmitted from one susceptible host to another, as commonly occurs, for example, within families, classrooms, and military populations. The compounds of the formula are also useful therapeutically in the treatment of rhinovirus infections in that the compounds are effective in reducing or blocking viral replication. For prophylactic or therapeutic treatment of rhinovirus infections,
Any anti-rinoviral effective amount of a compound of formula may be used. For therapeutic treatment, the dosage of the compound will depend primarily on the severity of the infection. For therapeutic or prophylactic treatment, the compound dosage varies from about 0.1 to 15 mg per kg of body weight of the patient or susceptible host.
Preferably, the compound dosage varies from about 1 mg/Kg to about 3 mg/Kg. Typically about 25 compounds
Unit doses containing 1 to 6 mg per day will produce the desired effect. With a suitable pharmaceutical carrier, compounds of the formula can be presented in solid dosage form such as tablets, capsules, powders, or suppositories. Powders can be administered orally or by insufflation. For the preparation of solid dosage forms, it is desirable to micronize the compound used.
In solid dosage form, the compounds can be carried in conventional carriers, such as binders such as gum arabic, corn starch, or gelatin, disintegrants such as corn starch, potato starch, or alginic acid, stearic acid or magnesium stearate. and inert fillers such as lactose, sucrose or corn starch. Compounds of the formula can be prepared using sterile liquids such as oil or water with the addition of pharmaceutically suitable surfactants or emulsifiers for oral, topical or parenteral administration.
Alternatively, it may be administered without addition as a liquid suspension or solution. A particularly suitable mode of administration is a liquid formulation of the compound applied directly to the nasal cavity, for example in the form of nasal sprays. Liquid formulations contain compounds such as peanut oil,
It is suitable to formulate with fixed oils such as sesame oil, cottonseed oil or olive oil. Peanut oil and sesame oil are particularly suitable for preparing formulations for intramuscular injection. Such oils can also be used in the preparation of soft gelatin-type and suppository formulations. Typically water, saline,
Aqueous dextrose and related sugar solutions, as well as glycerols such as polyethylene glycols, can be used to make liquid formulations. These formulations may suitably contain suspending agents such as methylcellulose, hydroxypropylcellulose or carboxymethylcellulose as well as buffers and preservatives. Exemplary of suitable pharmaceutical formulations are described below. Ketone compounds of the formula are available from Fieser and Fieser "Reagents for Organic Synthesis",
by treating one equivalent of the corresponding carboxylic acid derivative with two equivalents of the appropriate alkyl lithium, as generally described by John Willey & Sons, Inc., New York, p. 688 (1967). It will be done. The reaction is carried out in a solvent such as ether, tetrahydrofuran, p-dioxane, dimethoxyethane or diethylene glycol dimethyl ether at a temperature of -10 DEG C. to the reflux temperature of the solvent for 1/2 hour to 10 hours. Ketone compounds of formula _R
Appropriately R ₂ when has the meaning defined by Eq.
It can also be produced by the reaction of -O-substituted furancarboxylic acid derivatives with imidazolide derivatives. This reaction involves ether, tetrahydrofuran,
It is carried out in a solvent such as dioxane, dimethoxyethane or acetonitrile. The reaction mixture is first cooled to -10°C, after which the temperature is increased from about 25°C to the reflux temperature of the solvent, and the reaction time varies from about 0.5 to 10 hours. Imidazolide derivatives can be obtained by treating a suitably R ₂ -O-substituted furancarboxylic acid derivative with N,N'-carbonyldiimidazole or by treating a substituted carboxylic acid with thionyl _chloride . Substituted furoyl chlorides were prepared by HAStaab, Angew.Chem.Inhernat.Edit.1 volume.
351 (1962) by treatment with two equivalents of imidazole. Ketone compounds of the formula when the R ₂ -O- substituent is attached to the 5-position of the furan ring include a suitably R ₂ -O-substituted furan (wherein R ₂ is defined in the formula expression (with meaning)

[Formula] Friedel-Crafts acyl by the acyl halide of halo (wherein halo is a halogen, preferably chlorine or bromine, and R ₄ is a straight-chain or branched alkyl group of 1 to 4 carbon atoms) created by The reaction is carried out in the presence of an acid catalyst, such as boron trifluoride etherate, stannic chloride, zinc chloride, hydriodic acid or orthophosphoric acid, and optionally in the presence of a solvent, such as methylene chloride, nitromethane or benzene. It can be done. The suitable temperature for this reaction is -20℃
to the reflux temperature of the solvent, and the reaction time varies from about 0.5 to 10 hours. The R ₂ -O-substituted furan derivatives used herein are the thermal derivatives of suitably R ₂ -O-substituted Freud's acid (furanic acid or furocarboxylic acid) by procedures known in the art. (＞150
°C) obtained by decarboxylation. The R ₂ -O-substituted furancarboxylic acid derivatives used herein include the aromatic parent nuclei generally described on page 500 of the March reference cited above, as summarized below. Created by gender substitution. In the general reaction above, R ₂ has the meaning defined in the general formula, and L represents a leaving group such as nitro, fluoro, chloro, bromo, or iodo;
A preferred leaving group is chloro. Substituent L on the compound of structure 1 and R ₂ -O- on the compound of structure 2
The group is attached to the 3-, 4-, or 5-position of the furan ring. The above reaction can be carried out with or without a solvent. Suitable solvents for the reaction are chlorinated hydrocarbon solvents such as benzene, xylene, toluene, chlorobenzene, ethers such as bis(2-methoxyethyl)ether, 1,2-dimethoxyethane or anisole, dimethylformamide, dimethylacetamide. , 1-methyl-2-pyrrolidone or pyridine. Preferred solvents are xylene and dimethylacetamide. Copper metal or a copper salt such as cuprous chloride may optionally be added to the reaction. Suitable bases for the reaction include sodium or potassium metal, sodium hydride, potassium amide, potassium tert-butyrate, or other strong bases such as potassium carbonate, potassium hydroxide, sodium hydroxide and sodium carbonate. The reaction temperature is approximately 25
℃ to the reflux temperature of the solvent. Reaction times vary from about 1 hour to about 7 days. After the reaction is complete,
Treatment of carboxylate salt derivatives with mineral or organic acids yields compounds of structure 2. Alcohols represented by R2 _- OH that have utility in the above general reaction are either commercially available or prepared by reduction of the corresponding carboxylic acid or aldehyde. The following specific examples further illustrate the invention. Example Methyl 2-(5-tetradesiloxy)furyl ketone (A) 5-bromo-2-furanic acid 125.0 g (0.652 mol), 1-tetradecanol 210.0 g (0.978 mol), potassium tert-butoxide 183.0 g ( 1.630
mol) and 2500 ml of dimethylacetamide are heated with stirring. The tert-butanol formed during the reaction is distilled off and the mixture is then heated under reflux for 48 hours with stirring. Add 6 parts of ice water to the cooled mixture and acidify the mixture with malonic acid. The resulting precipitate was collected, dried and recrystallized twice from methanol, yielding 82.0 g (29%) of 5-(tetradesiloxy)-2-furanic acid.
will occur. Melting point 112-115°C (decomposition). (B) 5-(tetradesyloxy)-2-furanic acid 82.0
(0.253 mol), 41.0 g (0.253 mol) of N,N'-carbonyldiimidazole, and 800 ml of tetrahydrofuran are stirred at room temperature, during which carbon dioxide gas is evolved. When the reaction mixture was cooled to 0°C, N-[5-(tetradecyloxy)-
2-furoyl]imidazole. N-substituted imidazole in 500 ml of tetrahydrofuran
Cool 50.0 g (0.134 mol) in an ice bath. One equivalent of methylmagnesium bromide (50 ml of a 3M solution in ether) is slowly added to the stirred mixture over a period of 2 hours. The reaction was stirred for a further 3 hours, then an excess of 2NHC (500 ml) was added.
is added and the product is extracted with ether. The ether extract is separated, washed with water, dried over sodium sulfate, filtered and evaporated to dryness to yield methyl 2-(5-tetradesiloxy)furyl ketone. Melting point 70-72℃. Reference example 1 Methyl 5-(cis-9-octadecen-1-yloxy)-2furylketone 5-bromo-2-furanic acid 57.2g (0.300 mol), cis-9-octadecenol 121.0g (0.45
A mixture of 18.0 g (0.750 mol) of sodium hydride and 2 p-xylene is heated to reflux for 48 hours. The mixture is allowed to cool, then acidified with acetic acid and diluted with 2 portions of water. The organic layer is separated, dried and evaporated to dryness, and the residue is recrystallized from hexane to yield 5-(cis-9-octadecen-1-yloxy)-2-furanic acid. 5-(tetradecyloxy)- by the procedure of Example 1B
An appropriate amount of 5-(cis-9
Using -octadecen-1-yloxy)-2-furanic acid, methyl 5-(cis-9-octadecen-1-yloxy)-2-furyl ketone is obtained. Reference example 2 Ethyl 5-(9,12,15-octadecatriene-1-yloxy)-2-furylketone 5-bromo-2-furanic acid 57.0 g (0.300 mol), 9,12,15-octadecatrienol 119.0
g (0.450 mol) and potassium tert-butoxide 84
g (0.750 mol) in dry toluene are mixed with heating. The tert-butanol produced during the reaction was distilled off, and the mixture was heated to 110°C while stirring.
Reflux for 48 hours. The mixture is allowed to cool, then acidified with acetic acid and diluted with ice water. The toluene organic layer is separated, washed with water and extracted three times with 5% sodium bicarbonate solution. The combined aqueous extracts were cooled and acidified with 10% HC solution to give 5-(9,
12,15-octadecatriene-1-yloxy)-
Produces 2-furanic acid. Using the procedure of Example 1B, 5-(tetradecyloxy)
In place of -2-furanic acid, an appropriate amount of 5-(9,
12,15-octadecatriene-1-yloxy)-
Using 2-furanic acid and an appropriate amount of ethylmagnesium bromide in place of methylmagnesium bromide, ethyl 5-(9,12,15
-octadecatriene-1-yloxy)-2-furylketone is obtained. Example Measurement of anti-rinovirus activity Anti-rinovirus activity was measured according to the following procedure. Cells: HeLa cells susceptible to rhinovirus (R-
Hela) and human foreskin fibroblasts were grown in Eagle basal medium (BME) supplemented with 10% fetal bovine serum. Virus: Rhinovirus 39 (RV39) was used in most cases. Approx. original virus per cell
Adjustments were made by infecting at a multiplicity of 0.1 plaque forming units (PFU), a small volume was adsorbed for 90 min at 35°C, the cell sheet was washed with BME, fresh medium was added, and cultured at 35°C for 20–24 h. . Virus assay: Rhinovirus titers were determined by plaque method on R-HeLa cell sheets (80% rash) contained in 35 mm Petri dishes. Each assay was performed in duplicate. Add 0.1ml of virus to 35°C for 90 minutes.
4 ml of adsorbed to the cell layer and added magnesium.
of overlay medium (0.5 Methocel in BME) was used on each dish after the adsorption period. Infected cells were cultured at 35°C for 48 hours, then fixed with formalin and stained with crystal violet. Measurement of cytotoxicity: The cytotoxicity of the compound [methyl 2-(5-tetradecyloxy)furylketone] is (i)
Oyama VI and H.Eagle1956, Proc.Soc.Exp.Biol.
Med.91:305-307 and (ii) by measuring the total number of cells per culture medium after trypsin digestion. Measurement of compound binding to cells: Add 1 ml of BME to carbonyl cell culture medium in a 35 mm Petri dish at 35 °C.
were exposed to different concentrations of ¹⁴ C-labeled methyl 2-(5-tetradecyloxy)furyl ketone in At various times, the medium was removed from the culture medium and the cell sheets were washed three times with phosphate buffered saline pH 7.2. Cells from each culture were lysed in 1 ml of Hyamine Hydroxide (Sigma Chemical) and counted in a liquid scintillation spectrometer in toluene-based fluid. Results: Methyl 2-(5
-Tetradesyloxy)furylketone sensitivity is shown in Table 1. Virus was adsorbed to cells 1 hour before addition of BME containing the indicated concentrations of compounds.

Table: Several different human rhinoviruses were sensitive to the above compounds. The cytotoxicity of the above compounds is shown in Table 2. After culturing the cells with the above compound for 24 hours, the cell number and protein concentration were measured.

[Table] Table 3 shows the inhibition of RV39 when cells were pretreated with compounds (5 μg/ml).

[Table] Table 4 shows the antiviral effects when the above compounds were added after viral infection.

[Table] Formulation Example 1 Solution Methyl 2-(5-tetradesiloxy)furylketone 0.85g Alcohol 78.9ml Isopropyl myristate 5.0g Polyethylene glycol 400 10.0g Pure water Amount to make 100ml Alcohol, isopropyl myristate, and polyethylene glycol 400 together and dissolve the drug in it. Add enough pure water to make 100ml. Formulation Example 2 15,000 tablets Methyl 2-(5-dodecyloxy)furylketone 75g Lactose 1.216Kg Corn starch 0.3Kg The active ingredient, lactose, and corn starch are mixed uniformly. Granulate with 10% starch paste. Water content approx.
Dry to 2.5%. Sift through a No. 12 mesh sieve. Add and mix the following: Magnesium stearate 0.015Kg Corn starch 1.725Kg Compress with a suitable tablet press to a weight of 0.115g per tablet. Formulation Example 3 Soft gelatin capsules Methyl 2-(5-tetradesyloxy) Furyl ketone 0.25Kg Polysorbate 80 0.25Kg Corn oil Amount to make 25.0Kg Mix and fill into 50,000 soft gelatin capsules. Formulation Example 4 Powder % w/w Methyl 2-(5-tetradecyloxy) Furyl Ketone 1 Anhydrous silicon dioxide 0.5 Corn starch, lactose (same amount as fine powder) Sufficient amount

Claims (1)

[Claims] 1. An antirinovirus agent containing methyl 2-(5-tetradecyloxy)furyl ketone.