KR100438209B1 - Complex antimicrobial composition based on carvacrol, thymol, and citral - Google Patents

Abstract

PURPOSE: A natural antimicrobial composition containing carvacrol, thymol and citral as an effective ingredient is provided. It has an antimicrobial effect on bacteria and thus is useful for treatment and prevention of bacterial diarrhea, bronchial pneumonia or mastitis of animals. CONSTITUTION: The natural antimicrobial composition contains carvacrol, thymol and citral in a weight ratio of 1.0:0.2 to 0.8:0.1 to 0.2 and as an auxiliary component at least one selected from 0.01 to 1% by weight of sodium benzoate, 0.1 to 1.0% by weight of allantoin, 0.1 to 1% by weight of sodium propionate, 0.005 to 0.5% by weight of dexamethasone sodium phosphate, 0.01 to 0.1% by weight of tripelenamine hydrochloride, 1 to 10% by weight of brine and 1 to 10% by weight of propylene glycol.

Description

Complex antimicrobial composition based on carvacrol, thymol, and citral containing carbachol, thymol and citral as active ingredients

The present invention relates to a complex antimicrobial composition containing carbacrol, thymol and citral as an active ingredient, and more particularly, a certain ratio of carbacol, thymol and citral, which are plant extracts, are mixed. The present invention relates to an antimicrobial composition which is useful for the treatment and prevention of bacterial diarrhea, bronchial pneumonia or mastitis in animals.

At present, there is a lot of research to find a material to replace the chemical synthetic antimicrobial agent at the time when consumers' interest in organic foods or livestock products that do not contain antibiotics is increasing with the movement to limit the use of antibiotics for promoting growth. Therefore, carbacroll and thymol obtained from various plants mainly act on the cell wall of bacteria to denature and coagulate proteins in the cell wall structure, and also act on the cytoplasmic membrane to change the permeability of cations, which damages the reproductive process of the cell and eventually dies. [US 6,322,825 B1]. In particular, for parasitic pathogenic microorganisms, the infected cells are eliminated by themselves and serve to simultaneously inhibit cells and pathogenic microorganisms [US 6,322,825 B1]. In addition, carbacroll and thymol are known to have excellent anti-inflammatory effects [US 6,322,825 B1].

On the other hand, citral has a strong fragrance, strong antibacterial and anti-fungal action, is a component included in many foods everyday, and harmless to the human body licensed as a food additive [Kyungin Food and Drug Administration No. 101].

As a natural antibacterial agent currently being used and sold in the animal drug market, a mastitis agent using citrus is commercially available, and a cabacrol agent that kills intestinal pathogenic bacteria as an oral drug is commercially available in Korea. These products are all single-component formulations, which do not have perfect antimicrobial and anti-inflammatory effects and do not maintain proper aroma when used in feed. Currently, domestic antimicrobial products using citral have not been developed. Recently, it has been reported that cabacrol is effective in treating respiratory diseases due to bronchial dilatation [Boskabady, 2003].

On the other hand, where Staphylococcus aureus is a problem with the bacterial infection of the animals (Staphylococcus aureus) is a fast acquisition of resistance to antibiotics by strain infected animal in causing diarrhea and pneumonia antibiotics currently in use will not be treated. Therefore, we tried to develop a natural antimicrobial agent that has an excellent effect on the antibiotic resistance strain Staphylococcus aureus. In addition, Bodetella bronchiseptica, Mycoplasma hyopneumoniae , which is a respiratory pathogen in animals, and Salmonella typhimurium , which causes diarrhea in animals, Escherichia coli , etc. As a test strain, we tried to develop a new animal natural antimicrobial agent.

Therefore, the present inventors have studied to solve the above problems, and as a result, by mixing a certain ratio of carbacrolol, thymol and citral to develop a natural complex antimicrobial agent for use as an enteric infection, mastitis, disinfectant of animals It was completed.

Accordingly, an object of the present invention is to provide a natural composite antimicrobial composition for animals comprising citral, carbacroll and thymol in a constant ratio.

The present invention is characterized by an antimicrobial composition containing citral: carvacrol: thymol in a content of 1.0: 0.2 to 0.8: 0.1 to 0.2 (weight ratio).

Referring to the present invention in more detail as follows.

The present invention is a natural complex antimicrobial composition for animals newly mixed with a certain ratio of carvacrol (Carvacrol), thymol and citral plant extracts for the treatment and prevention of bacterial diarrhea, bronchial pneumonia or mastitis in animals Antimicrobial compositions useful in the present invention.

Animals in the present invention include livestock, including chickens, pigs, cattle, horses, dogs, ducks, goats and sheep; Freshwater fish, including trout, carp and eel; It also includes sea fish, including flatfish, rockfish, sea bream and shrimp.

First, citral, which is one of the active ingredients of the antimicrobial composition according to the present invention, is a natural substance extracted from grapefruit and various plants and has a molecular weight of 152.24, a colorless or pale yellow liquid, which has a lemon-like scent. use. In addition, it is a safe substance that can also be used as a mouthwash, and has an excellent effect on Gram-negative and Gram-positive bacteria and exhibits an antifungal effect. Aqueous solutions are insoluble in aqueous solutions, but are soluble in alcohol, and are used as mouthwashes and novel natural antimicrobial compositions. However, no product has been developed for animals.

In addition, among the active ingredients of the novel animal natural antimicrobial composition according to the present invention, carbacrol and thymol act to promote tissue regeneration, and these components are used as aromatherapy, and also have an immune boosting effect, so as to cure diseases and It will help prevention. In particular, carbacrol has been reported to have bronchial dilatation, which can be effectively used for respiratory therapy [Boskaboddy et al, 2003].

Therefore, the combination of citral, carbacroll and thymol is a novel functional novel natural complex antimicrobial agent having immunostimulatory action, bronchodilator action and anti-inflammatory action in addition to antibacterial action.

Particularly, in the present invention, it is preferable to mix citral, carbacroll and thymol in a weight ratio of 1.0: 0.2 to 0.8: 0.1 to 0.2, and when citral, carbacroll and thymol are less than 1.0: 0.2: 0.1 by weight, bronchial dilatation action When citral, carbacroll and thymol exceed 1.0: 0.8: 0.2 weight ratio, bronchial dilation and anti-inflammatory action increase, but odor stress reduces animal feed efficiency. have.

On the other hand, sodium benzoate (Sodium benzonate) that can be added as an auxiliary ingredient to the composition has an antiseptic effect and acts to increase the antimicrobial activity of the composition as a drug used as a pharmaceutical preservative. The preferred content of sodium benzoate is 0.01 to 1.0% by weight based on the total composition, there is a problem that the price is higher than the above range.

In addition, allantoin which can be added as an auxiliary ingredient to the composition (allantoin: 5-uraidhydantoin) is a substance produced by the decomposition of uric acid by uricase, and has a wound healing promoting action to revive cells of wound tissue. It synergizes with citral, further promoting inflammation repair. Preferred content of allantoin is 0.1 to 1.0% by weight based on the total composition, there is a problem that the toxicity of allantoin increases if the above range is exceeded.

In addition, sodium propionate (Sodium Propionate), which can be added as an auxiliary ingredient to the composition, acts as an antifungal substance to inhibit fungi causing mastitis. Its preferable content is 0.1 to 1.0% by weight based on the total composition, if the above range is exceeded there is a problem of increasing the toxicity due to propionic acid.

In addition, a steroidal anti-inflammatory agent having an anti-inflammatory effect can be added to the composition, and preferably 0.005 to 0.5% by weight of dexamethasone phosphate can be used with respect to the whole composition. At this time, if the above range is exceeded, there is a problem of residual.

In addition, an appropriate amount of antihistamine may be added to the composition for vascular contraction, secretion inhibition and sedation of the inflammatory site. As the antihistamines used in the composition, both antihistamines, which are effective and safe for humans and animals, may be used. Preferably, triplenmine hydrochloride may be included in an amount of 0.01 to 0.1 wt% based on the total composition. . At this time, if the above range is used, there is a problem of remaining when using inorganic.

In addition, a variety of bases suitable for infusion administration through nipples, including oily ointment, may be used as the base for the composition. As the oil-based ointment, petrolatum, vegetable oil can be used, and water-soluble base is used depending on the purpose of use. Preferably 1 to 10% by weight of brine or 1 to 10% by weight of propylene glycol can be used relative to the total composition. At this time there is a problem that the antimicrobial activity is lowered outside the range.

Such combinations are used to treat infectious diseases and disinfectants of resistant strains caused by the use of conventional antibiotics and chemotherapeutic agents.The treatment of infectious diseases is effective in preventing or disinfecting or treating intestinal infections, respiratory infections, and mastitis in animals. It is safe and can be used as a natural antimicrobial agent that can be used without distinction between inorganic and dry oil without requiring a separate drug holiday.

The present invention also includes compositions for the prevention and treatment of mastitis, which are effective in acute, chronic and subclinical mastitis as well as latent mastitis.

The co-formulation of the present invention may be administered in a variety of oral or parenteral formulations during actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used, may be used. Are prepared using.

Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose in lignans and lactone compounds and derivatives thereof. Mixed with gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solvents, emulsions, and syrups. In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.

Formulations for parenteral administration include sterile aqueous solutions, water-insoluble solvents, suspensions, emulsions, lyophilizers, suppositories. As the non-aqueous and suspending solvent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerol gelatin and the like can be used.

The content in the preparation of the active ingredient according to the present invention can be appropriately selected depending on the degree of absorption, inactivation rate, excretion rate, age and condition of the active ingredient in the body. The dosage of the co-formulation of the present invention is 10 to 100 mg / kg, preferably 20 to 50 mg / kg, and can be administered orally or intramammary injection once or three times a day.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1

Gram-positive strains of Staphylococcus aureus, KCCM40510, KCCM11593, KCCM41294, KCCM40881 and KCCM41291, which are gram-positive strains, were sold by the Korea Microorganism Conservation Center (KCTC), Muller-Hin. It was inoculated to and precultured at 37 ° C. The precultured strains were inoculated in Muller-Hinton medium to test the test medium, and the effect of the first antimicrobial activity on the ingredients shown in Table 1 was verified by the diffusion method using a paper disk. At this time, 60 μl of the components shown in the following Table 1 was adsorbed on the paper disc, placed on a test medium, and incubated overnight at 37 ° C. After 18 hours of incubation, the low-ring was measured in mm and summarized in Table 1 below.

As shown in Table 1, the antimicrobial activity in the composition of the composite 1 showed a superior effect than the citral, carbacroll, thymol, citral-carbacroll-thymol complex. From this result, it was confirmed that there is no antagonism between citral, carbacroll, and thymol.

Example 2

Natural antibacterial complex consisting of citral, carbacroll and thymol was treated as follows to observe the sterilization effect against the antimicrobial resistant strains. That is, the number of cultured strains was measured and 1 ml was inoculated into 8 ml of Mullertonton's liquid medium prepared in advance, and 1 ml of the natural antibacterial complex was added at the same time, 30 minutes, 60 minutes, 120 minutes and 24 hours before inoculation. The killing rate of the bacteria was measured. As controls, antibiotics ampicillin and nofloxacin, which are currently used in mastitis, were used. The number of bacteria was according to the National Committee for Clinical Laboratory Standards (NCCLS) method. The results are summarized in Table 2 to Table 6 below.

As shown in Tables 2 to 6, nofloxacin and ampicillin, the most commonly used antibiotics, were found to have no effect in treating staphylococcus aureus. In addition, the combination of carbacrol, thymol and citral showed superior antimicrobial effect than carbacrol, thymol and citral used alone. In particular, it was confirmed that the antimicrobial activity of the composite agent 1 is very excellent than the composite agent 2. Therefore, it can be seen that there is a difference in the antimicrobial activity according to the proper mixing ratio for these components.

Example 3

Antimicrobial susceptibility to Bodetella bronchiseptica, Mycoplasma hyopneumoniae , and Salmonella typhimurium , which cause diarrhea in animals, and Escherichia coli The test medium was inoculated in Muller-Hinton medium and precultured at 37 ° C. The precultured strains were inoculated in Müllertonton medium to make a test medium and compare the antimicrobial activity of the components shown in Table 7 below. 0.5 ml (1.0 × 10 ⁹ ) of the pre-cultured strain solution was added to 4 ml of M. Hinterton medium, and 0.5 ml of each of the ingredients shown in Table 7 was added thereto, and then the number of strains was confirmed after overnight culture. However, mycoplasma high opneumoniae was measured by liquid culture method due to the difficulty of culturing bacteria and slow growth. After 14 days, the growth of bacteria was measured by the change of medium color.

As shown in Table 7, the combination showed an excellent antimicrobial effect against the causative agents of respiratory and digestive infections. In particular, the combination of citral, carbacroll and thymol was superior to the mycoplasma bacterium, a causative agent of respiratory diseases, than the citral, carbacrolyl and thymol alone. In addition, the antimicrobial effect of Compound 1 was much better than that of Compound 2. Therefore, there was a significant difference in antimicrobial activity according to the mixing ratio of citral, carbacroll and thymol.

Example 4 Antimicrobial Activity of Natural Complex Antibiotic and Supplement

Following the addition of five auxiliary ingredients described in Table 8, the antimicrobial activity was measured in four strains.

As a result, the most excellent antimicrobial activity was shown in the combination No. 3 with 3% salt added as an auxiliary ingredient. In all five combinations tested, there was no decrease in antimicrobial activity by the auxiliary components.

Example 5: Somatic Reduction Test

A combination (citral: carbacro: thymol = 1: 0.7: 1.8) was added to two cows with mastitis three times at 12 hour intervals. 50 ml of the sample was taken before the injection of the drug to measure the somatic cell count (Cells / ml). Measurement of somatic cells was carried out with a somatic instrument of Fosmatic-90 (FOSSMATIC-90, Denmark).

As shown in Table 9, the natural complex antimicrobial agent showed an excellent somatic cell reduction effect, showing that the reduction of somatic cells in two cows with mastitis showed more than 90% after 36 hours.

Example 5: Toxicity Test

ICR mice (body weight: 25-30 g, 5 males per unit dose) were treated with 2,000 mg / Kg, 680 mg / Kg, 230 mg / kg combination (citral: carbachol: thymol = 1: 0.7: 0.18). Oral toxicity was administered once in the unit of Kg and 75 mg / Kg, and visually observed at intervals of two weeks, and mortality observation, general symptoms and weight measurements were performed for two weeks after the end of drug administration. Check the presence or absence.

As a result, the LD ₅₀ of the combination was found to be safe at 2,000 mg / kg or more, and there were no unusual symptoms at all doses, and the autopsy showed no change with the control.

Preparation Example 1: Preparation of Animal Oral Administration Powder

50 g of a natural complex antimicrobial agent (citral: carbacroll: thymol = 1: 0.7: 0.18) for the prevention and treatment of digestive organ infections for animals was mixed with 50 g of lactose, 20 g of crystalline cellulose, and 10 g of magnesium stearate. This mixture was mixed with feed excipients and formulated into pellet feeds of suitable size from 0.01 to 0.1 cm in diameter. The use was administered as a supplementary feed by mixing 500 g of the formulation with 1 ton of feed.

The components of the powder are as follows.

Active ingredient ··········· 50 g

50 g of lactose

20 g of crystalline cellulose

Magnesium Stearate10 g

Supplementary ingredients ·······

Excipients (powder) ······

Preparation Example 2 Preparation of Mastitis Injection

Natural complex antimicrobials (Citral: Carbacroll: Timol = 1.0: 0.7: 0.18) for the prevention and treatment of mastitis in cows are prescribed in the following formula by putting 7.5 g in one syringe into a dry or non-organic period. Infusion times.

The composition of the mastitis agent is as follows.

Natural complex antibacterial agent ... 1-5 g

Sodium benzoate 0.01-0.1 g

Allantoin 0.1 to 1 g

Sodium propionate 0.1 to 0.3 g

Tripereneamine hydrochloride ············· 0 to 0.02 g

Salt ·········· 3 g

Water-soluble or vegetable oils ·········

Distilled water balance

(100 g total)

Preparation Example 3 Preparation of Animal Respiratory Disease Treatment Injection

Natural complex antimicrobial agent (Citral: Carbachol: Timol = 1.0: 0.7: 0.18) for the prevention and treatment of respiratory tract infections for animals is injected through the nose or installed in an automatic control device indoors. Mounting. This is a device designed for disinfecting barn specialty, and it is easy to disinfect every corner of barn through micro-injection of chemicals, so it can control skin disease and various infectious diseases (foot-and-mouth disease, cholera, and obesque disease) in the early stage. In addition, by preventing the respiratory diseases of animals, etc., it is possible to prevent diseases and maintain a pleasant environment.

The composition ratio of the propellant is as follows.

Composite agent (citral: carbacro: thymol = 1.0: 0.7: 1,8) 1 to 5 g

Sodium benzoate 0.01-0.1 g

Ethyl Alcohol

Distilled water balance

As described above, the pharmaceutical composition according to the present invention acts directly on the bacteria causing the digestive tract infections, respiratory infections, and mastitis in animals to act to kill and significantly reduce the number of somatic cells. In addition, it exhibits an excellent effect on staphylococcus aureus resistant to antimicrobial agents and can increase the healing and prophylactic effects as a secondary selective fungicide against antimicrobial agents.

Claims (5)

Citral: Carvacrol: Carvacrol: Thymol is contained in a content of 1.0: 0.2-0.8: 0.1-0.2 (weight ratio). The composition according to claim 1, wherein 0.01 to 1% by weight of sodium benzonate, 0.1 to 1.0% by weight of allantoin, 0.1 to 1% by weight of sodium propionate, and 0.005 to 0.5% by weight of dexamethasone phosphate as auxiliary ingredients in the composition. %, 0.01 to 0.1% by weight of triprenamine hydrochloride, 1 to 10% by weight of brine, and 1 to 10% by weight of propylene glycol, wherein the composition of the animal natural complex antimicrobial composition . Orally administered drug comprising a natural complex antimicrobial composition of claim 1 or 2. An injection comprising a natural composite antimicrobial composition of claim 1 or 2. Disinfectant propellant comprising the natural complex antimicrobial composition of claim 1 or 2.