Anticoccidial compositions comprising ginkgo biloba leaves and uses thereof

Abstract

The present application relates to an anticoccidial composition comprising ginkgo biloba leaves and uses thereof. According to embodiments, the composition comprising ginkgo biloba leaves has an excellent effect of directly killing sporozoites that can induce coccidiosis, inhibiting penetration of sporozoites into cells, and / or inhibiting proliferation of sporozoites in cells, and has an excellent effect on prevention, improvement, and treatment of coccidiosis in vivo.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of priority based on Korean Patent Application No. 10-2020-0125244, filed on September 25, 2020, and the entire contents disclosed in the corresponding Korean patent application are incorporated in this specification. Technical Field

[0003] This application relates to an anticoccidial composition comprising ginkgo leaves and its use. Background Technology

[0004] Coccidiosis is an intestinal-associated disease caused by a protozoan parasite called *Eimeria*, belonging to the phylum *Apicomplexan*. Infection with coccidiosis causes digestive disorders, diarrhea, and weight loss, and can also lead to livestock death. Therefore, it has a significant economic impact on farms worldwide (Williams RB. A compartmentalized model for the estimation of the cost of coccidiosis to the world's chicken production industry, Int J Parasitol, 1999; 29(8): 1209-1229).

[0005] In recent years, many researchers have developed anticoccidial agents as therapeutics for coccidiosis. These anticoccidial agents, such as ionocarriers or chemically synthesized compounds, can prevent the formation of oocyst cell walls or asexual and sexual proliferation in protozoa. However, due to the long-term use of shuttle programs that alternately treat the resulting ionocarriers and chemically synthesized compounds, side effects such as the emergence of drug-resistant protozoa have occurred.

[0006] In particular, the accumulation of antibiotics in animals due to misuse and overuse is a serious problem, as humans ingest antibiotics through meat. Consequently, many countries around the world are banning the use of antibiotics due to antibiotic residues in livestock products. Therefore, there is an urgent need to develop and research alternatives to conventional anticoccidial agents, which exhibit side effects such as the emergence of resistant strains and internal residues.

[0007] Existing technology

[0008] Patent documents

[0009] (Patent Document 1) US Patent Publication No. 2008-0160000 Summary of the Invention

[0010] Technical issues

[0011] One embodiment of this application provides a feed composition for the prevention or relief of coccidiosis, comprising ginkgo leaves.

[0012] Other embodiments of this application provide a pharmaceutical composition for the prevention or treatment of coccidiosis, comprising at least one active ingredient selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and pharmaceutically acceptable salts thereof.

[0013] Other embodiments of this application provide a pharmaceutical composition for the prevention or treatment of coccidiosis, comprising ginkgo leaves.

[0014] Other embodiments of the present invention provide an antiprotozoan composition against Eimeria sp. protozoa, comprising at least one active ingredient selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide and their salts.

[0015] Other embodiments of this application provide an antiprotozoan composition against Eimeria protozoa, comprising ginkgo leaves.

[0016] Other embodiments of this application provide a method for preventing, alleviating, or treating coccidiosis, including the step of administering the composition to an animal other than a human.

[0017] Other embodiments of this application provide the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts in the preparation of compositions (e.g., pharmaceutical compositions) for the prevention, relief, and / or treatment of coccidiosis or in the preparation of antiprotozoan compositions; the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts in the prevention, relief, and / or treatment of coccidiosis; and / or the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts in the antiprotozoan activity against Eimeria protozoa (e.g., killing Eimeria protozoa; and / or inhibiting cell invasion and / or proliferation of Eimeria protozoa).

[0018] Other embodiments of this application provide the use of ginkgo leaves in the preparation of compositions (e.g., feed compositions, pharmaceutical compositions) for the prevention, relief and / or treatment of coccidiosis or in the preparation of antiprotozoan compositions; the use of ginkgo leaves for the prevention, relief and / or treatment of coccidiosis; and / or the use of ginkgo leaves for antiprotozoan drugs against Eimeria protozoa (e.g., killing Eimeria protozoa; and / or inhibiting cell invasion and / or proliferation of Eimeria protozoa).

[0019] Technical solution

[0020] Ginkgolide A is a compound with the molecular formula C 20 H 24 Compounds of O9, and having the following chemical formula 1 structure, such as CAS No. 15291-75-5, UNII-TAZ2DPR77B, CHEMBL465161, BN52020, DTXSID10873222 and / or HMS2089P12.

[0021] [Chemical Formula 1]

[0022]

[0023] Ginkgolide B is a compound with the molecular formula C 20 H 24 O 10 The compound, and having the structure of the following chemical formula 2, for example, it may be named Ginkgo lactone, and may be CAS No. 15291-77-7, BN52021 and / or AK160212.

[0024] [Chemical Formula 2]

[0025]

[0026] Ginkgolide C is a compound with the molecular formula C 20 H 24 O 11 The compound having the following chemical formula 3 structure, for example, CAS No. 15291-76-6, SCHEMBL16452771, MFCD02094178, ZINC85507063 and / or AKOS025311463.

[0027] [Chemical Formula 3]

[0028]

[0029] Quercetin is a substance with the molecular formula C 15 H 10Compounds of O7, having the structure of the following chemical formula 4, for example, may be named meletin, sophoretin and / or xanthaurine, and may be CAS No. 117-39-5.

[0030] [Chemical Formula 4]

[0031]

[0032] Kaempferol is a compound with the molecular formula C 15 H 10 Compounds of O6, having the structure of the following chemical formula 5, and for example, may be named Robigenin, Rhamnolutein, Populnetib, Trifolitin and / or Pelargidenolon, and may be CAS No. 520-18-3.

[0033] [Chemical Formula 5]

[0034]

[0035] Ginkgolide is a compound with the molecular formula C. 15 H 18 Compounds of O8, having the structure of the following chemical formula 6, and for example, it may be CAS No. 33570-04-6, UNII-M81D2O8H7U or CHEBI:3103.

[0036] [Chemical Formula 6]

[0037]

[0038] The compounds are selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and their salts. They can be purchased as commercial products, or obtained by extraction and separation from natural products, or prepared by commonly used organic synthesis methods, but are not limited thereto.

[0039] The salt of the compound in this application can refer to a physiologically acceptable salt of a substance in which the cation and anion are bound by electrostatic attraction, for example, a salt acceptable for feed compositions, a pharmaceutically acceptable salt, and / or a salt acceptable for antigenic animal compositions. For example, the salt can be at least one selected from the group consisting of metal salts, salts with organic bases, salts with inorganic acids, salts with organic acids, and salts with basic or acidic amino acids. In one embodiment, the metal salt can be at least one selected from the group consisting of alkali metal salts (sodium salts, potassium salts, etc.), alkaline earth metal salts (calcium salts, magnesium salts, barium salts, etc.), and aluminum salts; the salt with organic bases can be at least one selected from the group consisting of salts with triethylamine, pyridine, methylpyridine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylenediamine, etc.; and the salt with inorganic acids can be selected from hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid. At least one of the groups consisting of, etc.; the salt with organic acids may be at least one of the groups consisting of salts of formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.; the salt with basic amino acids may be at least one of the groups consisting of salts of arginine, lysine, ornithine, etc.; and the salt with acidic amino acids may be at least one of the groups consisting of salts of aspartic acid, glutamic acid, etc.

[0040] In this application, superior anticoccidiosis efficacy (activity, effect) may refer to at least one selected from the group consisting of (1) to (5) below (e.g., any one, two or more, three or more, or all of them):

[0041] (1) It had a higher anticoccidial index (ACI) compared with the control group;

[0042] (2) When administered to coccidiosis-induced animal subjects, the mortality rate, lesion score (e.g., appendiceal lesion score) and / or fecal oocyst excretion were reduced compared to the control group.

[0043] (3) It inhibits weight loss by inducing coccidiosis;

[0044] (4) Compared with the control group, it showed higher insecticidal activity against protozoa that induce coccidiosis; and

[0045] (5) Compared with the control group, the protozoa that induced coccidiosis had a higher inhibitory effect on cell invasion and a higher inhibitory effect on cell proliferation.

[0046] In this application, the control group may refer to a negative control group (a group without any treatment or a group treated with water and / or buffer) and / or a positive control group containing conventionally known anticoccidial agents (e.g., diclazuril, salinomycin, and / or gallic acid).

[0047] The composition according to one embodiment may have at least one feature selected from the group consisting of (1) to (6) below (e.g., one or more, two or more, three or more, four or more, five or more, or all six), and its features may be superior to those of the control group:

[0048] (1) Excellent anticoccidial activity;

[0049] (2) Excellent anti-antiprotozoan activity against protozoa that induce coccidiosis;

[0050] (3) Excellent acid resistance;

[0051] (4) Excellent heat resistance;

[0052] (5) Excellent in vivo stability and / or safety; and

[0053] (6) Excellent effect in relieving weight gain.

[0054] The composition according to one embodiment has excellent acid and / or heat resistance, thereby maintaining excellent anticoccidial activity for a long time when administered to the body, and has in vivo stability, thereby maintaining excellent anticoccidial activity even in environments with a variety of temperatures and / or a variety of pH ranges, and is suitable for a variety of products (e.g., feed additives), and its storage stability can be excellent.

[0055] According to one embodiment, the composition, when administered in vivo, is not absorbed by other tissues and organs (e.g., blood, liver, kidneys and / or spleen, etc.) except the intestine, thus leaving low residues in vivo, and therefore the composition can have excellent safety in vivo.

[0056] In one embodiment, the superior weight gain relief effect may refer to the superior effect of increasing the weight of the subject when the drug is administered to the subject, and in one embodiment, the weight gain may refer to daily weight gain, and the subject may be a subject in which coccidiosis has been induced in their body.

[0057] The composition according to one embodiment exhibits anticoccidioidomycin activity equivalent to or higher than conventionally known anticoccidial agents (e.g., sulfonamides such as sulfaquinoxaline, sulfachlorpyrifos and sulfadiazine, polyether ionocarrier antibiotics such as salinomycin and monensin sodium, amprolium, diclazuril, gallic acid and / or toltrazuril) without causing side effects or drug resistance, and can be safe for prolonged use because the composition contains a natural product (ginkgo leaf) with low toxicity or that does not remain in the body.

[0058] In this application, "prevention" refers to all actions that inhibit or delay the development of a disease by administering a composition according to one embodiment; "treatment" refers to all actions that alleviate or beneficially alter the symptoms of a suspected and diseased subject by administering a composition according to one embodiment; and "alleviation" refers to all actions that reduce parameters related to the condition of treating the disease by administering a composition according to one embodiment, such as, at least, the severity of symptoms. The disease may refer to coccidiosis.

[0059] In one embodiment, the composition comprises at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and their salts. The composition may be a composition containing ginkgo leaves, and the ginkgo leaves may be at least one selected from the group consisting of ginkgo leaf raw materials, dried materials, powdered materials, and extracts.

[0060] In another aspect, a feed composition comprising ginkgo leaves may be provided for the prevention or relief of coccidiosis.

[0061] The composition according to one embodiment may contain ginkgo leaves as an ingredient exhibiting anticoccidial activity. The ginkgo leaves may be at least one (e.g., any one, two or more, three or more, or all) selected from the group consisting of ginkgo leaf raw materials, dried materials, powdered materials, and extracts.

[0062] Ginkgo leaves can refer to the leaves of the ginkgo tree (Ginkgo biloba) belonging to the Ginkgoaceae family. Unlike most other seed plant leaves, ginkgo leaves are fan-shaped, split in the middle, and in one embodiment, the leaves may be blue (green) in spring and / or yellow with autumn foliage in autumn. Ginkgo leaves have been used as medicine in traditional Chinese medicine for a long time, and in modern pharmaceuticals, ginkgo leaf extracts are used as therapeutic agents for cerebral and peripheral blood flow degeneration, sensory nervous system diseases, memory and cognitive function decline, and have been reported to have anti-cancer inhibitory effects.

[0063] In one embodiment, the ginkgo leaf may refer to a blue leaf (green leaf) preceding the yellow autumn leaf or a yellow leaf accompanying the yellow autumn leaf.

[0064] Ginkgo leaf raw material can refer to unprocessed (e.g., dried, ground into powder and / or extracted) ginkgo leaves.

[0065] Ginkgo leaf dried material can be obtained by drying ginkgo leaf raw materials, and can be obtained by methods such as vacuum drying, fluidized bed drying, room temperature drying and / or freeze drying.

[0066] Ginkgo leaf powder can be obtained by grinding and / or pulverizing raw ginkgo leaves and / or dried ginkgo leaves, and can be obtained using a pulverizer.

[0067] In one embodiment, ginkgo leaf powder can be prepared by drying ginkgo leaves at 50°C to 90°C for 4 to 24 hours and then grinding them into powder using a pulverizer.

[0068] In this application, "extract" refers to a preparation obtained by squeezing a herb containing a suitable extract and concentrating the extract by evaporation. It can be a dried product obtained by drying the extract obtained through extraction, a diluted solution or concentrate of the extract, its preparation material, and / or purification material. Ginkgo leaf raw material can be prepared using commonly known extraction, separation, and purification methods in the art. Specifically, extraction methods such as boiling water extraction, hot water extraction, floral fragrance extraction, reflux cooling extraction, or ultrasonic extraction can be used.

[0069] In one embodiment, the ginkgo leaf extract can be commercially available or prepared using extraction methods and solvents known in the pharmaceutical or food industries. For example, conventional extraction methods may include solvent extraction, ultrasonic extraction, filtration, and / or reflux extraction. In one embodiment, the ginkgo leaf extract can be prepared by extraction with an extraction solvent or by fractionation by adding a fractionating solvent to an extract prepared by extraction with an extraction solvent. For example, the ginkgo leaf extract can be extracted using a solvent selected from the group consisting of water, straight-chain or branched alcohols having 1 to 4 carbon atoms, propylene glycol, butanediol, glycerol, acetone, ethyl acetate, butyl acetate, chloroform, diethyl ether, dichloromethane, hexane, and mixtures thereof. Specifically, the alcohol having 1 to 4 carbon atoms can be ethanol, methanol, isopropanol, propanol, butanol, and / or tert-butanol. Extraction can be performed by adding an extraction solvent in an amount up to 1 to 10 times the weight of the dried ginkgo leaves; specifically, extraction can be performed by adding an extraction solvent in an amount of 2 to 3 times the weight of the dried ginkgo leaves. The extraction temperature can be 30℃–100℃, 50℃–90℃, or 60℃–80℃. Furthermore, the extraction time can be 10 hours–48 hours, 12 hours–30 hours, or 18 hours–24 hours. Additionally, extraction can be repeated 1 to 5 times, 2 times, or 4 times.

[0070] In this application, "coccidiosis" refers to a disease caused by coccidia (protozoa that can induce coccidiosis, such as Eimeria sp.) parasitizing the cytoplasm of the submucosal tissue of the digestive tract epithelium and damaging the epithelium, thus causing enteritis. It is also a protozoan disease that causes economic losses due to weight gain and regression caused by soft stools, diarrhea, and bloody stools, leading to prolonged market age in broiler farms. Coccidiosis can occur not only in broilers but also in birds and mammals. Specifically, it can infect cattle, rabbits, goats, dogs, cats, and laboratory animals such as mice and rats. Particularly, it can cause fatal harm to poultry such as chickens. In one embodiment, coccidiosis may include acute coccidiosis, subacute coccidiosis, and chronic coccidiosis. Acute coccidiosis can manifest as bloody stools, energy loss, and anemia within 48 hours of infection, and infected subjects may die. Subacute coccidiosis can manifest as hemorrhagic diarrhea and / or anemia after infection. Chronic coccidiosis can manifest as loose stools and / or weight loss after diarrhea within 1 to 2 days after infection.

[0071] When the oocysts (cysts, eggs) of coccidial protozoan species mature into sporocysts with spores under high humidity and high temperature, the oocysts are infectious. They are easily transmitted and their life cycle is repeatedly repeated when they are excreted in feces after a certain life cycle within the body of a subject. It is known that the oocytes (cysts) of coccidial protozoan species are highly resistant to the external environment, with the cyst wall consisting of an inner and outer layer. The outer layer of the cyst wall is a gelatinous material that strongly resists external physical pressure, while the inner layer is rich in nucleoproteins, thus strongly resisting chemical irritants such as disinfectants. An oocyst of a coccidial protozoan species can contain four sporangia, each of which can contain two sporozoites. After infecting an animal in the form of oocysts, these sporangia are released as sporangia and sporozoites, multiplying within the cell. The sporozoites, through sexual and / or asexual reproduction, can form oocysts, which are then extracted in feces. In one implementation, the sporozoite can be used to mean the same thing as a protozoan, and the sporozoite (protozoan) can cause lesions.

[0072] According to one implementation, coccidiosis can be caused by protozoa of the genus Eimeria. In one embodiment, the Eimeria protozoa can be selected from Eimeria acervulina, Eimeria tenella, Eimeria maxima, Eimeria venomaecatrix, Eimeria brunetti, Eimeria hagani, Eimeria mitis, Eimeria praecox, Eimeria mivati, Eimeria aurati, Eimeria baueri, Eimeria lepidosirenis, Eimeria leucisci, Eimeria rutile, and Eimeria van dendritic. Eimeria vanasi, Eimeria amphisbaeniarum, Eimeria witchery, Eimeria yemenensae, Eimeria adenoeides, Eimeria colchici, Eimeria curvata, Eimeria dispersa, Eimeria duodenalis, Eimeria fraterculae, Eimeria gallopavonis, Eimeria innocua, Eimeria meleagridis, Eimeria meleagrimitis, Eimeria phasiani, Eimeria pusse Eimeria purpureicephali, Eimeria ahsata, Eimeria alabamensis, Eimeria aliyeviEimeria alijevi, Eimeria aspheronica, Eimeria arloingi, Eimeria arundeli, Eimeria bakuensis, Eimeria bovis, Eimeria cameli, Eimeria caprina, Eimeria caprovina, Eimeria christenseni, Eimeria clethrionomyis, Eimeria coecicola, Eimeria contorta, Eimeria couesii, Eimeria clitorella Eimeria crandallis, Eimeria dammahensis, Eimeria dowleri, Eimeria exigua, Eimeria falciformis, Eimeria farasanii, Eimeria ferrisi, Eimeria flavescens, Eimeria gallatii, Eimeria granulosa, Eimeria hirci, Eimeria intestinalis, Eimeria irresidua, Eimeria intricata, Eimeria jolchijevi, Eimeria crandallis, Eimeria dammahensis, Eimeria dowleri, Eimeria exigua, Eimeria falciformis, Eimeria farasanii, Eimeria ferrisi, Eimeria flavescens, Eimeria gallatii, Eimeria granulosa, Eimeria hirci, Eimeria intestinalis, Eimeria irresidua, Eimeria intricata, Eimeria jolchijevi, Eimeria crandallis, Eimeria exigua ... Eimeria krijgsmanni, Eimeria larimerensis, Eimeria macusaniensis, Eimeria magna, Eimeria marconii, Eimeria media, Eimeria melanuri, Eimeria misiEimeria myoxi, Eimeria nagpurensis, Eimeria nieschulzi, Eimeria inakohlyakimovae, Eimeria ovinoidalis, Eimeria pallida, Eimeria palustris, Eimeria papillata, Eimeria perforans, Eimeria phocae, Eimeria pileata, Eimeria pipistrellus, Eimeria piriformis, Eimeria proterimi At least one of the following groups: Eimeria procyonis, Eimeria punctate, Eimeria roobroucki, Eimeria saudiensis, Eimeria sealanderi, Eimeria separata, Eimeria stiedae, Eimeria ursini, Eimeria vermiformis, Eimeria weybridgensis, Eimeria wobati, and Eimeria zuernii.

[0073] The composition according to one embodiment can have excellent effects in preventing, alleviating and / or treating coccidiosis caused by at least one protozoan selected from the group consisting of at least one Eimeria genus protozoan described in Table 1 below, which can cause coccidiosis in the animals described in Table 1 respectively.

[0074] Table 1

[0075]

[0076]

[0077]

[0078] The composition according to one embodiment can be highly effective in preventing, alleviating and / or treating coccidiosis caused by Eimeria tenella, Eimeria spp. and / or Eimeria gianti.

[0079] In one implementation, prevention or mitigation of coccidiosis may refer to at least one of the following groups (e.g., any one, two or more, three or more, or all of them) selected from (1) to (4) below, and for example, compared with the control group (negative control group and / or positive control group), at least one of the following groups selected from (1) to (4) below may be reduced, suppressed, and / or increased:

[0080] (1) Reduce at least one of the groups consisting of disease score (e.g., appendiceal disease score), fecal oocyst excretion and mortality;

[0081] (2) Inhibit weight loss caused by coccidiosis;

[0082] (3) Increase the anticoccidial index (ACI); and

[0083] (4) Reduce cell invasion of Eimeria protozoa, proliferation of protozoa in cells, or both.

[0084] In one embodiment, the lesion scoring method for determining the lesion score can refer to the literature of Johnson JK and Reid WM (Joyce Johnson and W. Malcolm Reid, Anticoccidial drugs: Lesion scoring techniques in battery and floor-pen experiments with chickens), Experimental Parasitology, 1970), and the lesion score can be 0 to 4 degrees. In one embodiment, the lesion score can refer to the lesion score measured in the appendix, duodenum, and / or jejunum, and it can be calculated by summing the lesion scores measured in each organ (appendix, duodenum, and / or jejunum).

[0085] In one implementation, the amount of fecal ovules extracted can be measured by collecting feces from the subject and using a microscope or a counting chamber (e.g., a McMaster chamber).

[0086] In one implementation, the mortality rate may refer to the mortality rate of animal subjects who have been induced with coccidiosis, and the number of subjects who died from causes other than coccidiosis can be excluded by performing an autopsy.

[0087] In one embodiment, the weight of a subject induced with coccidiosis may be less than that of a subject without coccidiosis, and the composition according to one embodiment can inhibit weight loss by inducing coccidiosis.

[0088] In one embodiment, the anticoccidial index can be calculated according to Equation 1 below, and in Equation 1, the lesion score can be calculated according to the aforementioned method.

[0089] (Equation 1)

[0090] Anticoccidial Index (ACI) = (Survival rate after challenge inoculation (%)) + (Daily weight gain compared to negative control group (%)) - (Lesion score × 10) - (Fecal oocyst excretion index)

[0091] The challenge inoculation can refer to the administration (e.g., oral inoculation) of a protozoan capable of inducing coccidiosis. In one embodiment, the survival rate can be the survival rate measured on days 5–10, 7–10, 8–10, 7–9, 7–8, or 7 after the challenge inoculation, and can be measured by excluding the number of subjects who died from causes other than coccidiosis through autopsy.

[0092] The weight gain in Equation 1 above compared to the negative control group can be a percentage calculated based on the values ​​of the negative control group (e.g., a negative control group of uninfected protozoa).

[0093] The lesion score in Equation 1 above is as described above.

[0094] In Equation 1 above, the percentage is calculated based on the values ​​of the negative control group (e.g., the negative control group of infected protozoa). When the calculated value is above 0% and below 1%, the fecal oocyst excretion index can be 0; when the calculated value is above 1% and below 26%, the fecal oocyst excretion index can be 5; when the calculated value is above 26% and below 51%, the fecal oocyst excretion index can be 10; when the calculated value is above 51% and below 76%, the fecal oocyst excretion index can be 20; and when the calculated value is above 76% and below 100%, the fecal oocyst excretion index can be 40.

[0095] In one embodiment, the active ingredient (at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts) may be included in the feed composition in the following amounts: less than 1 w / w%, less than 1 w / w%, 10 -1 w / w% or less, 5×10 -2 w / w% or less, 2.5×10 -2 w / w% or less, 2×10 -2 w / w% or less, 1.25×10 -2 w / w% or less, 10 -2 w / w% or less, 9×10 -3 w / w% or less, 8×10 -3 w / w% or less, 7×10 -3 w / w% or less, 6×10 -3 w / w% or less, 5×10 -3 w / w% or less, 4×10 -3 w / w% or less, 10 -7 w / w% or more, 10 -6 w / w% or more, 10 -5 w / w% or more, 10 -4 w / w% or more, 5×10 -4 w / w% or more, 10 -3 w / w% or more, 1.5×10 -3 w / w% or more, 2×10 -3 w / w% or more, 3×10 -3 w / w% or more, 4×10 -3 w / w% or more, 5×10 -3 w / w% or more, 10 -7 w / w%~1w / w%, 10 -7 w / w% ~10 -1 w / w%, 10 -7 w / w% ~ 5 × 10 -2 w / w%, 10 -7 w / w% ~10 -2 w / w%, 10 -7 w / w% ~ 5 × 10 -3 w / w%, 10 -7 w / w% ~ 4 × 10 -3 w / w%, 10 -7 w / w% ~10 -3 w / w%, 10 -7 w / w% ~ 5 × 10 -4 w / w%, 10 -7 w / w% ~10 -4 w / w%, 10-7 w / w%~10 -5 in / in%、10 -6 in / in%~1in / in%、10 -6 w / w%~10 -1 in / in%、10 -6 w / w%~5×10 -2 in / in%、10 -6 w / w%~10 -2 in / in%、10 -6 w / w%~5×10 -3 in / in%、10 -6 w / w%~4×10 -3 in / in%、10 -6 w / w%~10 -3 in / in%、10 -6 w / w%~5×10 -4 in / in%、10 -6 w / w%~10 -4 in / in%、10 -6 w / w%~10 -5 in / in%、10 -5 in / in%~1in / in%、10 -5 w / w%~10 -1 in / in%、10 -5 w / w%~5×10 -2 in / in%、10 -5 w / w%~10 -2 in / in%、10 -5 w / w%~5×10 -3 in / in%、10 -5 w / w%~4×10 -3 in / in%、10 -5 w / w%~10 -3 in / in%、10 -5 w / w%~5×10 -4 in / in%、10 -5 w / w%~10 -4 in / in%、10 - 4 in / in%~1in / in%、10 -4 w / w%~10 -1 in / in%、10 -4 w / w%~5×10 -2 in / in%、10 -4 w / w%~10 -2 in / in%、10 -4 w / w%~5×10 -3 in / in%、10 -4w / w% ~ 4 × 10 -3 w / w%, 10 -4 w / w% ~10 -3 w / w%, 10 -4 w / w% ~ 5 × 10 -4 w / w%, 10 -3 w / w%~1w / w%, 10 -3 w / w% ~10 -1 w / w%, 10 -3 w / w% ~ 5 × 10 -2 w / w%, 10 -3 w / w% ~10 -2 w / w%, 10 -3 w / w% ~ 5 × 10 -3 w / w%, 10 -3 w / w% ~ 4 × 10 -3 w / w%, 10 -3 w / w% ~ 2×10 - 3 w / w% or 10 -3 w / w% ~ 1.5 × 10 -3 w / w%. In one embodiment, the feed composition may be a feed containing an active ingredient within a range based on total weight (e.g., a mixed feed and / or compound feed ultimately ingested by the animal).

[0096] In one embodiment, the active ingredient (at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts) may be included in the feed composition at the following concentrations: less than 10,000 ppm, less than 5,000 ppm, less than 2,000 ppm, less than 1,000 ppm, less than 500 ppm, less than 400 ppm, less than 300 ppm, less than 250 ppm, less than 200 ppm, less than 125 ppm, less than 125 ppm, less than 100 ppm, less than 90 ppm, less than 80 ppm, less than 70 ppm, less than 65 ppm, less than 60 ppm, less than 50 ppm, less than 40 ppm, 0.001 ppm. Above, above 0.01ppm, above 0.1ppm, above 1ppm, above 5ppm, above 10ppm, above 15ppm, above 20ppm, above 30ppm, above 40ppm, above 50ppm, 0.001ppm~1000ppm, 0.001ppm~500ppm, 0.001ppm ~300ppm, 0.001ppm~200ppm, 0.001ppm~125ppm, 0.001ppm~100ppm, 0.001ppm~90ppm, 0.001ppm~80ppm, 0.001ppm~70ppm, 0.001ppm~60ppm, 0.001ppm~ 50ppm, 0.001ppm~40ppm, 0.001ppm~30ppm, 0.003ppm~1000ppm, 0.003ppm~500ppm, 0.003ppm~300ppm, 0.003ppm~200ppm, 0.003ppm~125ppm, 0.003ppm ~100ppm, 0.003ppm~90ppm, 0.003ppm~80ppm, 0.003ppm~70ppm, 0.003ppm~60ppm, 0.003ppm~50ppm, 0.003ppm~40ppm, 0.003ppm~30ppm, 0.01ppm~1000 ppm, 0.01ppm~500ppm, 0.01ppm~300ppm, 0.01ppm~200ppm, 0.01ppm~125ppm, 0.01ppm~100ppm, 0.01ppm~90ppm, 0.01ppm~80ppm, 0.01ppm~70ppm, 0.01 ppm~60ppm, 0.01ppm~50ppm, 0.01ppm~40ppm, 0.01ppm~30ppm, 0.1ppm~1000ppm, 0.1ppm~500ppm, 0.1ppm~300ppm, 0.1ppm~200ppm, 0.1ppm~125ppm, 0.1ppm~100ppm, 0.1ppm~90ppm, 0.1ppm~80ppm, 0.1ppm~70ppm, 0.1ppm~60ppm, 0.1ppm~50ppm, 0.1ppm~40ppm , 0.1ppm~30ppm, 1ppm~1000ppm, 1ppm~500ppm, 1ppm~300ppm, 1ppm~200ppm, 1ppm~125ppm, 1ppm~100ppm, 1pp m~90ppm, 1ppm~80ppm, 1ppm~70ppm, 1ppm~60ppm, 1ppm~50ppm, 1ppm~40ppm, 1ppm~30ppm, 3ppm~1000ppm, 3p pm~500ppm, 3ppm~300ppm, 3ppm~200ppm, 3ppm~125ppm, 3ppm~100ppm, 3ppm~90ppm, 3ppm~80ppm, 3ppm~70ppm , 3ppm~60ppm, 3ppm~50ppm, 3ppm~40ppm, 3ppm~30ppm, 5ppm~1000ppm, 5ppm~500ppm, 5ppm~300ppm, 5ppm~20 0ppm, 5ppm~125ppm, 5ppm~100ppm, 5ppm~90ppm, 5ppm~80ppm, 5ppm~70ppm, 5ppm~60ppm, 5ppm~50ppm, 5ppm~4 0ppm, 5ppm~30ppm, 10ppm~1000ppm, 10ppm~500ppm, 10ppm~300ppm, 10ppm~200ppm, 10ppm~125ppm, 10ppm~1 00ppm, 10ppm~90ppm, 10ppm~80ppm, 10ppm~70ppm, 10ppm~60ppm, 10ppm~50ppm, 10ppm~40ppm or 10ppm~30ppm. .

[0097] In one embodiment, when the active ingredient (selected from at least one of the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and their salts) is included within the above range, the anticoccidial activity may be superior compared to the case where the active ingredient is not included within the above range.

[0098] In one embodiment, the ginkgo leaves may be included in the feed composition in the following amounts: less than 1 w / w%, less than 1 w / w%, 10 -1 w / w% or less, 5×10 -2 w / w% or less, 2.5×10 -2 w / w% or less, 2×10-2 w / w% or less, 1.25×10 -2 w / w% or less, 10 -2 w / w% or less, 9×10 -3 w / w% or less, 8×10 -3 w / w% or less, 6.25×10 -3 w / w% or less, 7×10 -3 w / w% or less, 6×10 -3 w / w% or less, 5×10 -3 w / w% or less, 4×10 -3 w / w% or less, 10 - 7 w / w% or more, 10 -6 w / w% or more, 10 -5 w / w% or more, 10 -4 w / w% or more, 5 × 10 -4 w / w% or more, 10 -3 w / w% or more, 1.5 × 10 -3 w / w% or more, 2 × 10 -3 w / w% or more, 3 × 10 -3 w / w% or more, 4 × 10 -3 w / w% or more, 5 × 10 - 3 w / w% or more, 6.25 × 10 -3 w / w% or more, 10 -7 w / w% to 1w / w%, 10 -7 w / w% ~ 10 -1 w / w%, 10 -7 w / w% ~ 5 × 10 -2 w / w%, 10 -7 w / w% ~ 10 -2 w / w%, 10 -7 w / w% ~ 5 × 10 -3 w / w%, 10 -7 w / w% ~ 4 × 10 -3 w / w%, 10 -7 w / w% ~ 10 -3 w / w%, 10 -7 w / w% ~ 5 × 10 -4 w / w%, 10 -7 w / w% ~ 10 -4 w / w%, 10 -7 w / w% ~ 10 - 5 w / w%, 10 -6 w / w% to 1w / w%, 10 -6w / w%~10 -1 in / in%、10 -6 w / w%~5×10 -2 in / in%、10 -6 w / w%~10 -2 in / in%、10 -6 w / w%~5×10 -3 in / in%、10 -6 w / w%~4×10 -3 in / in%、10 -6 w / w%~10 -3 in / in%、10 -6 w / w%~5×10 -4 in / in%、10 -6 w / w%~10 -4 in / in%、10 -6 w / w%~10 -5 in / in%、10 -5 in / in%~1in / in%、10 -5 w / w%~10 -1 in / in%、10 -5 w / w%~5×10 -2 in / in%、10 -5 w / w%~10 -2 in / in%、10 -5 w / w%~5×10 -3 in / in%、10 -5 w / w%~4×10 -3 in / in%、10 -5 w / w%~10 -3 in / in%、10 -5 w / w%~5×10 -4 in / in%、10 -5 w / w%~10 -4 in / in%、10 -4 in / in%~1in / in%、10 -4 w / w%~10 -1 in / in%、10 -4 w / w%~5×10 -2 in / in%、10 -4 w / w%~10 -2 in / in%、10 -4 w / w%~5×10 -3 in / in%、10 -4 w / w%~4×10 -3 in / in%、10 -4 w / w%~10 -3 in / in%、10 -4w / w% ~ 5 × 10 -4 w / w%, 10 -3 w / w%~1w / w%, 10 -3 w / w% ~10 -1 w / w%, 10 -3 w / w% ~ 5 × 10 -2 w / w%, 10 -3 w / w% ~10 -2 w / w%, 10 -3 w / w% ~ 5 × 10 -3 w / w%, 10 -3 w / w% ~ 4 × 10 -3 w / w%, 10 -3 w / w% ~ 2×10 -3 w / w% or 10 -3 w / w% ~ 1.5 × 10 -3 w / w%. In one embodiment, the feed composition may be a feed containing an active ingredient within a range based on total weight (e.g., a mixed feed and / or compound feed ultimately ingested by the animal).

[0099] In one embodiment, the ginkgo leaves may be included in the feed composition at the following concentrations: less than 10,000 ppm, less than 5,000 ppm, less than 2,000 ppm, less than 1,000 ppm, less than 500 ppm, less than 400 ppm, less than 300 ppm, less than 250 ppm, less than 200 ppm, less than 125 ppm, less than 125 ppm, less than 100 ppm, less than 90 ppm, less than 80 ppm, less than 70 ppm, less than 65 ppm, less than 60 ppm, less than 50 ppm, less than 40 ppm, more than 0.001 ppm, more than 0.01 ppm, more than 0.1 ppm, more than 1 ppm, more than 5 ppm, more than 10 ppm. 15ppm or more, 20ppm or more, 30ppm or more, 40ppm or more, 50ppm or more, 0.001ppm~1000ppm, 0.001ppm~500ppm, 0.001ppm~300ppm, 0.001ppm~200ppm, 0.001ppm~125ppm, 0.0 01ppm~100ppm, 0.001ppm~90ppm, 0.001ppm~80ppm, 0.001ppm~70ppm, 0.001ppm~60ppm, 0.001ppm~50ppm, 0.001ppm~40ppm, 0.001ppm~30ppm, 0.003pp m~1000ppm, 0.003ppm~500ppm, 0.003ppm~300ppm, 0.003ppm~200ppm, 0.003ppm~125ppm, 0.003ppm~100ppm, 0.003ppm~90ppm, 0.003ppm~80ppm, 0.00 3ppm~70ppm, 0.003ppm~60ppm, 0.003ppm~50ppm, 0.003ppm~40ppm, 0.003ppm~30ppm, 0.01ppm~1000ppm, 0.01ppm~500ppm, 0.01ppm~300ppm, 0.01ppm ~200ppm, 0.01ppm~125ppm, 0.01ppm~100ppm, 0.01ppm~90ppm, 0.01ppm~80ppm, 0.01ppm~70ppm, 0.01ppm~60ppm, 0.01ppm~50ppm, 0.01ppm~40ppm, 0. 01ppm~30ppm, 0.1ppm~1000ppm, 0.1ppm~500ppm, 0.1ppm~300ppm, 0.1ppm~200ppm, 0.1ppm~125ppm, 0.1ppm~100ppm, 0.1ppm~90ppm, 0.1ppm~80ppm, 0.1ppm~70ppm, 0.1ppm~60ppm, 0.1ppm~50ppm, 0.1ppm~40ppm, 0.1ppm~30ppm, 1ppm~1000ppm, 1ppm~500 ppm, 1ppm~300ppm, 1ppm~200ppm, 1ppm~125ppm, 1ppm~100ppm, 1ppm~90ppm, 1ppm~80ppm, 1ppm~70ppm, 1ppm~60ppm, 1ppm~50ppm, 1ppm~40ppm, 1ppm~30ppm, 3ppm~1000ppm, 3ppm~500ppm, 3ppm~300ppm, 3pp m~200ppm, 3ppm~125ppm, 3ppm~100ppm, 3ppm~90ppm, 3ppm~80ppm, 3ppm~70ppm, 3ppm~60ppm, 3ppm~50p pm, 3ppm~40ppm, 3ppm~30ppm, 5ppm~1000ppm, 5ppm~500ppm, 5ppm~300ppm, 5ppm~200ppm, 5ppm~125pp m, 5ppm~100ppm, 5ppm~90ppm, 5ppm~80ppm, 5ppm~70ppm, 5ppm~60ppm, 5ppm~50ppm, 5ppm~40ppm, 5ppm~ 30ppm, 10ppm~1000ppm, 10ppm~500ppm, 10ppm~300ppm, 10ppm~200ppm, 10ppm~125ppm, 10ppm~100ppm , 10ppm~90ppm, 10ppm~80ppm, 10ppm~70ppm, 10ppm~60ppm, 10ppm~50ppm, 10ppm~40ppm or 10ppm~30ppm. .

[0100] In one embodiment, when ginkgo leaves are included in the above-mentioned range, their anticoccal activity can be superior compared to ginkgo leaves included outside the above-mentioned range.

[0101] In this application, "feed" may refer to any natural or artificial diet, meal, or component of a meal that is intended for animal consumption, ingestion, digestion, or suitability thereto. In a feed composition according to one embodiment, concentrated feed and / or specialty feed may be further included. Concentrated feed is a byproduct obtained through the purification of seeds, fruits, and grains, including grains such as wheat, oats, and corn, and may be bran, including rice bran, wheat bran, and barley bran; sesame cake, a byproduct obtained from pressing oil from soybeans, liquids, sesame seeds, flax seeds, coconuts, etc.; and residues such as residual starch, the main component of starch residue, which is the remainder after starch removal from sweet potatoes and potatoes; fish-solubles, obtained by concentrating fresh liquids obtained from fishmeal, fish waste, and fish; animal feed, such as dried whey, wherein whey is the remainder after the production of casein from meat meal, blood meal, feather meal, skim milk powder, cheese milk, skim milk, etc.; yeast, chlorella, and / or seaweed, etc.

[0102] According to one embodiment, a feed composition can refer to feed in the form ultimately ingested by animals, dietary supplements and / or feed additives that can be mixed with feed. Dietary supplements are compositions, for example, containing preparations that provide therapeutic or digestive aids to animals, and can refer to compositions that are not a common source of calories, i.e., an energy source, but are ingested outside of normal animal feed. Feed additives are substances added to feed for various purposes, such as supplementing nutrition and preventing weight loss, improving the digestibility of fiber in feed, improving oil quality, preventing reproductive disorders and enhancing fertility, and preventing summer heat stress. In one embodiment, it can refer to substances added for the purpose of preventing, alleviating, or treating coccidiosis.

[0103] In one embodiment, the feed composition may be a feed additive, and when the feed additive according to one embodiment is mixed into feed (e.g., mixed feed and / or compound feed ultimately ingested by the animal), it may be at a concentration of 0.001% (w / w) or more, 0.005% (w / w) or more, 0.01% (w / w) or more, 0.05% (w / w) or more, 0.1% (w / w) or more, or 0% (w / w) or more, based on the total weight of the feed. Above 0.5% (w / w), below 1% (w / w), below 0.5% (w / w), below 0.1% (w / w), below 0.05% (w / w), below 0.01% (w / w), below 0.005% (w / w), 0.001%~1% (w / w), 0.001%~0.5% (w / w), 0.001%~0.1% (w / w), 0.001%~0.05% (w / w), 0. 001%~0.01% (w / w), 0.001%~0.005% (w / w), 0.005%~1% (w / w), 0.005%~0.5% (w / w), 0.005%~0.1 % (w / w), 0.005% ~ 0.05% (w / w), 0.005% ~ 0.01% (w / w), 0.01% ~ 1% (w / w), 0.01% ~ 0.5% (w / w), 0.01 It can be added at a weight of 0.1% to 0.1% (w / w), 0.01% to 0.05% (w / w), 0.05% to 1% (w / w), 0.05% to 0.1% (w / w), 0.1% to 1% (w / w), 0.1% to 0.5% (w / w), or 0.5% to 1% (w / w), and according to one embodiment, it can be mixed with feed ingredients, supplements, adjuvants, and / or other kinds of additives, excluding the active ingredient.

[0104] In one embodiment, when the feed additive is included within the above-mentioned range, its anticoccidial activity may be superior compared to that of feed additives included outside the above-mentioned range.

[0105] On the one hand, a pharmaceutical composition for the prevention or treatment of coccidiosis can be provided, comprising at least one (e.g., any one, two or more, three or more, or all of) selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and their pharmaceutically acceptable salts as active ingredients.

[0106] In another aspect, a pharmaceutical composition comprising ginkgo leaves may be provided for the prevention or treatment of coccidiosis.

[0107] The pharmaceutical composition according to one embodiment can be used as a single formulation and can also be used as a mixed formulation by further comprising an approved pharmaceutical composition known to have preventive or therapeutic effects on coccidiosis. A unit dosage form of the drug can be prepared by adding a pharmaceutically acceptable carrier, excipient, or diluent.

[0108] In this application, "pharmaceuticalally acceptable" means having no significant irritant effect on organisms and not inhibiting the biological activity and properties of the drug-active substance. A pharmaceutical composition comprising a pharmaceutically acceptable carrier according to one embodiment may have any one of the following formulations selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories.

[0109] The pharmaceutical composition can be a variety of oral or parenteral formulations. In the case of formulations, it can be prepared using commonly used fillers, expanders, binders, wetting agents, disintegrants, diluents such as surfactants or excipients.

[0110] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc. These solid dosage forms can be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., into one or more compounds. In addition to simple excipients, lubricants such as magnesium stearate, talc, etc., can be used. Liquid dosage forms for oral administration, besides the commonly used simple diluents water and liquid paraffin, can also contain suspensions, internal solutions, emulsions, syrups, etc., as well as various excipients such as wetting agents, sweeteners, flavorings, preservatives, etc.

[0111] Preparations for parenteral administration can include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solutions and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate can be used. As a base for suppositories, polyethylene glycol fatty acid esters, Tween 61, cocoa butter, lauryl acetate, and glycerin gelatin can be used.

[0112] In one embodiment, the pharmaceutical composition can be formulated into various forms for use according to common methods for various purposes, such as oral preparations like powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, injectable preparations like sterile injectable solutions, and can be administered orally or via various routes including intravenous administration, intraperitoneal administration, subcutaneous administration, rectal administration, and local administration.

[0113] In one embodiment, the pharmaceutical composition may further include a carrier, excipient, or diluent, and examples of suitable carriers, excipients, or diluents may include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. Furthermore, the pharmaceutical composition may also additionally include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc.

[0114] In one embodiment, the effective amount of the active ingredient (e.g., at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their pharmaceutically acceptable salts) or ginkgo leaf in the pharmaceutical composition may vary depending on the patient's (subject's) age, sex, and weight, generally 0.0001 mg / kg to 0.001 mg / kg, 0.0001 mg / kg to 0.01 mg / kg, 0.0001 mg / kg to 0.1 mg / kg, 0.0001 mg / kg to 1 mg / kg, 0.0001 mg / kg to 10 ... mg / kg~250mg / kg, 0.0001mg / kg~500mg / kg, 0.0001mg / kg~1000mg / kg, 0.001mg / kg~0.01mg / kg, 0.001mg / kg~0.1mg / kg, 0.001mg / kg~1mg / kg, 0.001m g / kg~10mg / kg, 0.001mg / kg~100mg / kg, 0.001mg / kg~250mg / kg, 0.001mg / kg~500mg / kg, 0.001mg / kg~1000mg / kg, 0.01mg / kg~0.1mg / kg, 0.01mg / kg~1 mg / kg, 0.01mg / kg~10mg / kg, 0.01mg / kg~100mg / kg, 0.01mg / kg~250mg / kg, 0.01mg / kg~500mg / kg, 0.01mg / kg~1000mg / kg, 0.1mg / kg~1mg / kg, 0.1mg / kg~10mg / kg, 0.1mg / kg~100mg / kg, 0.1mg / kg~250mg / kg, 0.1mg / kg~500mg / kg, 0.1mg / kg~1000mg / kg, 1mg / kg~10mg / kg, 1mg / kg~100mg / kg, 1mg / kg~25 The dosage per kg of body weight is 0 mg / kg, 1 mg / kg~500 mg / kg, 1 mg / kg~1000 mg / kg, 10 mg / kg~100 mg / kg, 10 mg / kg~250 mg / kg, 10 mg / kg~500 mg / kg, 10 mg / kg~1000 mg / kg, 100 mg / kg~250 mg / kg, 100 mg / kg~1000 mg / kg, 250 mg / kg~500 mg / kg, 250 mg / kg~1000 mg / kg, or 500 mg / kg~1000 mg / kg, and can be administered daily or every other day, or divided into 1 to 3 doses daily.However, since the dosage can be increased or decreased depending on the route of administration, disease severity, sex, weight, age, etc., this dosage does not limit the scope of this application in any way. In one embodiment, when the composition is administered intraperitoneally, the composition can be administered at a concentration of 0.001 mg / kg to 50 mg / kg.

[0115] In one embodiment, the dosage of the pharmaceutical composition can be within a range depending on factors such as the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity.

[0116] In one embodiment, the concentration of ginkgo leaves contained in the active ingredient (e.g., at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and pharmaceutically acceptable salts thereof) or pharmaceutical composition is as described above.

[0117] In one embodiment, the pharmaceutical composition can be administered to a subject via multiple routes. Administration can refer to providing a substance to a subject (patient) by any suitable method. The route of administration of the pharmaceutical composition can be oral and / or parenteral, through all common routes, as long as the target tissue is reached. In the case of parenteral administration, options include external application to the skin, intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, and / or intrapleural injection. Furthermore, the composition according to one embodiment can be administered using any device capable of delivering the active ingredient to target cells.

[0118] One aspect may provide an antiprotozoan composition against Eimeria protozoa, comprising at least one (e.g., any one, two or more, three or more, or all) selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts as an active ingredient.

[0119] On the other hand, an antiprotozoan composition containing ginkgo leaves can be provided, which is an antiprotozoan of the genus *Eimeria*. The *Eimeria* protozoan is as described above.

[0120] In one embodiment, the antiprotozoan activity (effect, efficacy) against Eimeria protozoa is superior, which may refer to the following characteristics (1) and / or (2), for example, it may exhibit the following characteristics (1) and / or (2) compared with control groups (negative control group and / or positive control group):

[0121] (1) It has excellent killing effects on Eimeria protozoa; and / or

[0122] (2) Inhibit the cell invasion of Eimeria protozoa and / or inhibit the proliferation of protozoa in cells.

[0123] In one embodiment, at least one ingredient selected from the group consisting of ginkgolides A, B, C, quercetin, kaempferol, ginkgolide, and their salts, or ginkgo leaves, may be included in the antigenic animal composition within the aforementioned concentration range in the feed composition and / or pharmaceutical composition. In this embodiment, compositions containing active ingredients within the aforementioned concentration range may exhibit superior antigenic animal activity compared to compositions containing active ingredients outside the aforementioned concentration range.

[0124] On the other hand, a method for preventing, alleviating, or treating coccidiosis can be provided, comprising the step of administering the composition (e.g., a feed composition, feed additive, pharmaceutical composition, and / or antiprotozoan composition) to an animal. In one embodiment, prior to administering the composition, the method may further include identifying (selecting) a subject (patient) who requires prevention, alleviation, or treatment of coccidiosis. The composition and coccidiosis are as described above. According to one embodiment, identifying the subject may include detecting oocysts of protozoa capable of inducing coccidia in feces isolated from the subject.

[0125] In a method for preventing, alleviating, or treating coccidiosis according to one embodiment, the method of administration, route of administration, and / or dosage of the composition are as described above.

[0126] According to one embodiment, the composition can be administered at a pharmaceutically effective dose. In this application, a "pharmaceutically effective dose" refers to an amount sufficient to treat a disease under a reasonable benefit / risk ratio suitable for medical treatment, and the effective dose level can be determined based on the patient's disease type, severity, drug activity, drug sensitivity, timing of administration, route of administration and excretion ratio, treatment duration, concurrently used drug-containing components, and other components well known in the medical field. According to one embodiment, the composition can be administered as a standalone therapeutic agent or in combination with other anticoccidial agents, and can be administered simultaneously with, separately from, or sequentially with conventional therapeutic agents, and can be administered once or multiple times. Taking all factors into account, it is important to administer an amount that achieves maximum effect with minimal dosage without side effects, and this can be easily determined by those skilled in the art.

[0127] In one embodiment, the subjects to which the methods for preventing, mitigating, or treating coccidiosis are applied are animals that have or may have coccidiosis. These animals may be mammals, including humans, horses, cattle, mice, rats, dogs, cats, etc., birds, fish, amphibians, and / or reptiles, including poultry (e.g., breeder chickens, broilers, and / or laying hens).

[0128] In one embodiment, the animal used in the method for preventing, alleviating, or treating coccidiosis may be at least one selected from the group consisting of the animals described in Table 1 above, such as at least one selected from the group consisting of humans, chickens, ducks, geese, turkeys, quails, pheasants, pigeons, parrots, cattle, pigs, goats, sheep, horses, antelopes, eland, monkeys, cats, dogs, mice, rats, rabbits, raccoons, squirrels, bats, guinea pigs, camels, llamas, alpacas, wombats, lizards, goldfish, crucian carp, tilapia, barramundi, lungfish, and European whitefish. In one embodiment, the animal may be an animal other than a human.

[0129] On the other hand, the invention provides the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts in the preparation of compositions (e.g., pharmaceutical compositions) or antiprotozoal compositions for the prevention, relief, and / or treatment of coccidiosis; the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts for the prevention, relief, and / or treatment of coccidiosis; and / or the use of at least one selected from the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgolide, and their salts for antiprotozoal (e.g., killing Eimeria protozoa; and / or inhibiting cell invasion and / or proliferation of Eimeria protozoa) against Eimeria protozoa.

[0130] On the other hand, there is a use of Ginkgo biloba leaves in the preparation of compositions (e.g., feed compositions, pharmaceutical compositions) or antiprotozoan compositions for the prevention, relief and / or treatment of coccidiosis; a use of Ginkgo biloba leaves for the prevention, relief and / or treatment of coccidiosis; and / or a use of Ginkgo biloba leaves for antiprotozoan activity against Eimeria protozoa (e.g., killing Eimeria protozoa; and / or inhibiting cell invasion and / or proliferation of Eimeria protozoa).

[0131] In the above-mentioned uses, the selected material is at least one of the group consisting of ginkgolide A, ginkgolide B, ginkgolide C, quercetin, kaempferol, ginkgol, and their salts, and the ginkgo leaves, coccidiosis, antigenic animals, and Eimeria protozoa are as described above.

[0132] Beneficial effects

[0133] A composition containing ginkgo leaves according to one embodiment has excellent direct killing effect on protozoa capable of inducing coccidiosis, has cell invasion inhibition effect on protozoa and / or cell proliferation inhibition effect on protozoa, and has excellent preventive, alleviating and therapeutic effects on coccidiosis in vivo. Attached Figure Description

[0134] Figure 1 The anticoccidial index (ACI) is shown after Eimeria tenella challenge inoculation based on treatment with Ginkgo biloba powder and anticoccidial agents (dicrazine, salinomycin).

[0135] Figure 2 The study demonstrated the inhibitory effects of treatment with Ginkgo biloba leaf powder and anticoccidial agents (diclazuril, salinomycin, and gallic acid) on cell invasion and proliferation of Eimeria tenella. Detailed Implementation

[0136] The invention will be described in more detail below through the following embodiments. However, the embodiments are for illustrative purposes only, and the scope of the invention is not limited by these embodiments.

[0137] Example 1: In vivo anticoccidial activity of Ginkgo biloba leaf powder

[0138] Example 1-1. Experimental Facilities

[0139] An in vivo anticoccidial efficacy study was conducted at an animal testing facility in Gyeongsangnam-do, South Korea. One-day-old female Ross broilers were individually weighed and randomly assigned to groups for the experiment. The experimental design considerations and conditions are described in Table 2.

[0140] Table 2

[0141] category Experimental variables Aquaculture type cage Broiler chicken free-range age 1 day old Total experimental period 22 days gender female Number of broilers / cage 30 Number of repetitions / treatment groups 2 repetitions Number of treatment groups 6 groups Total number of broiler chickens 360 broiler chickens Challenges in inoculating protozoan species tender Eimeria coccidia Challenge: Number of ovarian cysts inoculated 10,000 oocysts orally inoculated into broilers

[0142] The farm was managed according to the Korean Poultry Farming Management Guidelines. The cages and the farm were cleaned and disinfected before the trial began. The farm was maintained at a temperature of 40°C–41°C and a humidity of 40%–50%, and this was continuously monitored.

[0143] Examples 1-2. Preparation of Ginkgo Leaf Powder

[0144] Ginkgo leaves were obtained and used from Cheongoksan Farm in Mitan-myeon, Pyeongchang-gun, Gangwon-do, and Jecheon Bank Farm in Susan-myeon, Jecheon-si, Chungcheongbuk-do. Green ginkgo leaves produced between July and September were dried in a 60°C dryer for 10 hours and then ground into a fine powder using a 100-mesh pulverizer to prepare ginkgo leaf powder, which was used in later experiments.

[0145] Examples 1-3. Experimental Design

[0146] The feed used was from Feed A1-choi (Korea). Various ingredients, including diclazuril (Yuhan Corporation YUHANDICLA product), salinomycin (Cheil Bio Cheilsalino-60 product), and gallic acid (Sigma product), along with the ginkgo leaf products prepared in Examples 1-2, were added to the feed and self-mixed at the concentrations described in Table 3. No antibiotics or additives were used in either the general feed or the mixed feed, and no anticoccidial agents were added except for the individual ingredients. Throughout the experiment, the broilers were fed freely. Each control or experimental group consisted of 30 one-day-old broilers randomly placed in cages and fed a general feed (A1-choi product) for 7 days. The prepared mixed feed was then divided into control and experimental groups for consumption.

[0147] The feed formulations used in the control group (negative control group or positive control group) and the experimental group, as well as whether coccidiosis was induced by Eimeria tenella, are described in Table 3 below.

[0148] Oral vaccination (challenge vaccination) was administered to 14-day-old broilers, with each chicken receiving 10,000 oocysts (more than 90% of which are sporozoites of Eimeria tenella) to induce coccidiosis.

[0149] Table 3

[0150] Group deal with Uninfected negative control group General feed Negative control group of infection Eimeria tenella infection + regular feed Positive control group 1 (salinomycin treatment group) Eimeria tenella infection + salinomycin 60ppm Positive control group 2 (diclazuril treatment group) Eimeria tenella infection + Diclazuril 1ppm Ginkgo leaf powder treatment group Eimeria tenella infection + Ginkgo biloba powder 125ppm

[0151] Examples 1-4. Determination of the anticoccidial activity of Ginkgo biloba leaf powder

[0152] The anticoccidial effect of the experimental groups designed in Examples 1-3 above is expressed as the anticoccidial index (ACI), which is calculated using the following Equation 2. The ACI score is out of 200. The higher the ACI score, the better the anticoccidial ability. When the ACI score is above 120 and below 140, it is determined to be effective as an anticoccidial material. When the ACI score is above 140 and below 160, it is determined to be excellent as an anticoccidial material. When the ACI score is above 160, its anticoccidial effect is determined to be very excellent (Luis Miguel De Pablos et al., Anticoccidial activity of maslinic acid against infection with Eimeria tenella in chickens, Parasitol Res, 2010).

[0153] (Equation 2)

[0154] Anticoccidial Index (ACI) = (Survival rate after challenge inoculation (%)) + (Daily weight gain compared to negative control group (RWG, %)) - (Lesion score × 10) - (Fecal oocyst excretion index)

[0155] 1) Survival rate: The number of dead individuals was recorded daily, and post-mortem examinations were performed to determine the cause of death, excluding individuals who died from causes other than coccidiosis. The survival rate (%) from challenge inoculation to day 8 was used to calculate the anticoccidial index.

[0156] The survival rates of the control and experimental groups are shown in Table 4 below.

[0157] Table 4

[0158] Group Survival rate (%) Uninfected negative control group 97 Negative control group of infection 90 Salinomycin treatment group 93 Diclazuril treatment group 97 Ginkgo leaf powder treatment group 100

[0159] Furthermore, the mortality rate of ginkgo leaf powder was 0% during the 4-week feeding period, and no damage to major organs (kidneys, spleen, liver, and heart) was identified, thus ensuring the safety of ginkgo leaf powder at a level of 125 ppm.

[0160] 2) Daily weight gain: The weight of individuals in each cage was measured before and on day 7 after protozoan challenge vaccination. The difference was divided by the number of days to calculate the daily weight gain (ADG, g / day). "Daily weight gain compared to the negative control group (RWG, %)" was calculated by dividing the daily weight gain (ADG, average daily gain; g / day) of each experimental group by the weight gain (ADG, g / day) of the uninfected negative control group, and then multiplying by 100. This was used to calculate the anticoccidial index.

[0161] Daily weight gain (ADG, g / day) measured in each control and experimental group, and daily weight gain (RWG, %) compared to the negative control group, are shown in Table 5 below.

[0162] Table 5

[0163]

[0164] 3) Lesion Scoring: On day 8 post-challenge inoculation, necropsy was performed on 4 broilers per cage, and the intestines were cut open. Each coccidial lesion in the appendix region of the broilers was scored. The lesion scoring method followed the literature of Johnson JK & Reid WM (1970) (Anticoccidial drugs: Lesion scoring techniques in battery and floor-pen experiments with chickens, Experimental Parasitology, 1970). The lesion score ranged from 0 to 4, with 0 corresponding to a normal appendix, 1 to mild infection, 2 to moderate infection, 3 to severe infection, and 4 to very severe infection or death. The measured appendix lesion score was multiplied by 10 to calculate the lesion index, which was used to calculate the anticoccidial index.

[0165] The appendiceal lesion scores measured in each control and experimental group are described in Table 6 below.

[0166] Table 6

[0167] Group Appendicitis score lesion index Uninfected negative control group 0.02 0 Negative control group of infection 3.05 31 Salinomycin treatment group 2.7 27 Diclazuril treatment group 2.88 29 Ginkgo leaf powder treatment group 1.68 17

[0168] 4) Fecal oocyst excretion: All feces from cages on days 6-8 of challenge inoculation were collected, mixed, and randomly sampled 3 times, 1g each time. After flocculating the oocysts in 1g of feces with saline, the oocyst excretion was measured using a McMaster chamber. The results are shown in Table 7 below.

[0169] Table 7

[0170] Group Oocyte excretion volume / g Ovum cyst index Uninfected negative control group 0 Negative control group of infection 1.9.E+08 40 Salinomycin treatment group 1.3.E+08 20 Diclazuril treatment group 9.5.E+07 10 Ginkgo leaf powder treatment group 1.1.E+08 20

[0171] The oocyst excretion rate (%) compared to the infected negative control group was calculated by dividing the oocyst excretion rate of each group by the oocyst excretion rate of the infected negative control group and multiplying by 100. The oocyst excretion rate index was calculated as follows: 0% to less than 1% compared to the infected negative control group; 5% to less than 26%; 10% to less than 26%; 20% to less than 51%; 20% to less than 76%; and 40% to less than 100%.

[0172] As described above, the anticoccidial index of each experimental group measured by Equation 2 is shown in Table 8 below. As shown in Table 8, the anticoccidial index of the Ginkgo biloba leaf powder treatment group was higher than that of the positive control group, diclazuril, or salinomycin treatment group, thus demonstrating excellent anticoccidial efficacy.

[0173] Table 8

[0174] Group Anticoccidial Index (ACI) Uninfected negative control group 196 Negative control group of infection 101 Salinomycin treatment group 141 Diclazuril treatment group 148 Ginkgo leaf powder treatment group 162

[0175] As confirmed in Tables 4-7 above, compared to the uninfected negative control group, coccidiosis developed in the challenge-inoculated *Eimeria tenella*-infected negative control group, resulting in decreased survival rate and weight gain, and increased lesion index and fecal oocyst excretion. Compared to the infected negative control group, survival rate and weight gain were increased, while lesion index and fecal oocyst excretion were decreased in the salinomycin-treated group and the diclazuril-treated group used as an anticoccidial control. Compared to the infected negative control group, survival rate and weight gain were improved, while lesion index and fecal oocyst excretion were decreased in the ginkgo leaf powder-treated group. This is a superior effect compared to salinomycin 60 ppm and diclazuril 1 ppm.

[0176] As described above, the anticoccidial index of each experimental group measured by Equation 2 above was... Figure 1 As shown in Table 8 above. Figure 1 As shown in Table 8, it can be observed that the anticoccidial index of the Ginkgo biloba leaf powder treatment group was higher than that of the positive control group, diclazuril or salinomycin treatment group, thus showing excellent anticoccidial efficacy.

[0177] Example 2. Direct killing effect of Ginkgo biloba leaf powder against Eimeria protozoa.

[0178] In this embodiment, the direct protozoan (sporophyte) killing ability of three representative Eimeria species known to infect most farms (Eimeria tenella, Eimeria spurula, and Eimeria giant) was assessed.

[0179] A certain amount of oocysts from various protozoa were placed in test tubes containing glass beads and crushed. The crushed oocyst cell walls and other debris were then removed. The internal sporangia were purified using a Percoll density gradient and washed with PBS solution. Sporangia from *Eimeria tenella*, *Eimeria squarrosa*, and *Eimeria giantiformis* were treated with reagents containing sodium taurocholate (Sigma-Aldrich, USA) and trypsin (Gibco, USA) respectively for decapacitation, incubated, and then washed once with PBS solution to obtain the protozoa.

[0180] Ginkgo leaf powder was reacted with anticoccidial agents, salinomycin, diclazuril, and gallic acid (hereinafter, materials) at different concentrations ranging from 1 to 500 ppm with three species of Eimeria protozoa. Only live protozoa (sporophytes) were counted under a microscope. The mortality rate (%) of protozoa treated with each material was then measured compared to a PBS-treated negative control group, and the minimum concentration required to directly kill 50% of the protozoa is shown in Table 9 below.

[0181] Table 9

[0182]

[0183] As shown in Table 9 above, in the gallic acid treatment group, gallic acid at concentrations up to 500 ppm did not have a 50% direct killing effect on protozoa capable of inducing coccidia. Similarly, in the case of diclazuril, diclazuril at concentrations up to 500 ppm did not have a 50% direct killing effect on *Eimeria tenella* and *Eimeria giantiformis* protozoa. In Ginkgo biloba leaf powder (green and yellow leaves), at significantly lower concentrations than other groups, 50% of *Eimeria tenella* and *Eimeria giantiformis* were killed, thus confirming an excellent protozoic killing effect.

[0184] Example 3. Inhibitory effect of Ginkgo biloba leaf powder on cell invasion and intracellular proliferation of Eimeria coccidia protozoa.

[0185] In this embodiment, the inhibitory effect of Ginkgo biloba powder on intracellular protozoan invasion and proliferation was studied using the MDBK cell line, a representative animal cell line known to cause infection and proliferation of Eimeria spp.

[0186] Example 3-1. Inhibitory effect of Ginkgo biloba leaf powder treatment on protozoan cell invasion.

[0187] 100,000 MDBK cells (purchased from ATCC) were aliquoted into 24-well plates and incubated at 37°C for 12 hours. Protozoa of *Eimeria tenella* were obtained similarly to the method described in Example 2 above. 200,000 protozoa were added to each well of the cell-allocated cells, and the cells were treated with each material (ginkgo leaf powder and anticoccidial agent, salinomycin, diclazuril, and gallic acid) at a concentration (0.5–10 ppm) and cultured at 41°C for 24 hours. The negative control group consisted of MDBK cells infected with *Eimeria tenella* protozoa, and the positive control group consisted of cells incubated with *Eimeria tenella* protozoa in solutions of salinomycin, diclazuril, or gallic acid. Subsequently, the cells were washed twice with PBS to remove uninvaded protozoa. After removing cells and intracellular protozoa using a pipette, DNA was extracted from the cells and PCR was performed using primers specific to the Eimeria tenella ITS-1 (Internal Transcriptional Spacer-1) gene. The sequences of the primers used are described in Table 10 below.

[0188] Table 10

[0189]

[0190] The Ct values ​​of each material before and after washing were compared and corrected with the ΔCt value of the negative control group to calculate the cell invasion inhibition rate (%) of each material on protozoa. The results are shown in [Figure Number]. Figure 2 and Table 11 below.

[0191] Example 3-2. Inhibitory effect of Ginkgo biloba leaf powder on intracellular proliferation of protozoan cells.

[0192] 100,000 MDBK cells (purchased from ATCC) were aliquoted into 24-well plates and incubated at 37°C for 12 hours. Eimeria tenella protozoa were obtained similarly to the method described in Example 2 above. 200,000 protozoa were added to each well of the cell-allotted plates, and the cells were treated with each material (ginkgo leaf powder and anticoccidial agent, salinomycin, diclazuril, and gallic acid) at the specified concentrations, and further cultured at 41°C for 24 hours. The negative control group consisted of MDBK cells infected with protozoa, and the positive control group consisted of cells incubated with Eimeria tenella protozoa in salinomycin and diclazuril solutions. After removing the cells and protozoa within them using a pipette, DNA was extracted from the cells and PCR was performed using primers specific to the Eimeria tenella ITS-1 (Internal Transcriptional Spacer-1) gene. The sequences of the primers used are described in Table 10 above.

[0193] The Ct values ​​before and after washing for each material were compared to calculate the cell invasion inhibition rate (%) and intracellular protozoan proliferation inhibition rate (%) of each material-treated protozoan. The results were... Figure 2 As shown in Table 11 below.

[0194] Table 11

[0195]

[0196]

[0197] like Figure 2 As shown in Table 11 above, to confirm the inhibition rates of cell invasion and intracellular protozoan proliferation of *Eimeria tenella* protozoa, Ginkgo biloba powder at 5 ppm and 10 ppm showed inhibition rates of over 40% in both cell invasion and intracellular protozoan proliferation. The 10 ppm concentration of Ginkgo biloba powder showed superior inhibitory effects on cell invasion and intracellular protozoan proliferation compared to salinomycin and diclazuril. Compared to gallic acid, it showed lower inhibitory effects on cell invasion but significantly higher inhibitory effects on intracellular protozoan proliferation. Commercially available anticoccidial agents, diclazuril and gallic acid, showed only one effect on either cell invasion or intracellular protozoan proliferation inhibition for each protozoan, while salinomycin and Ginkgo biloba powder showed both effects, with Ginkgo biloba powder showing better inhibitory effects on intracellular protozoan proliferation than salinomycin.

[0198] Example 4. Evaluation of anticoccidial efficacy of ginkgo leaf powder based on its components.

[0199] To confirm the components of Ginkgo biloba leaf powder that exhibit anticoccal activity, the mortality rate of *Eimeria tenella* protozoa was evaluated by treatment with ginkgolides A, B, C, quercetin, kaempferol, ginkgolide, or Ginkgo biloba leaf powder. The concentration of each material was selected from 5 ppm to 500 ppm, and the protozoan mortality rate (%) was measured using a method similar to that described in Example 2 above, as shown in Table 12 below.

[0200] Table 12

[0201]

[0202] Table 12 above confirms that Ginkgo biloba leaf powder at concentrations above 16 ppm killed 50% of *Eimeria tenella* protozoa, and at concentrations above 63 ppm killed 100% of *Eimeria tenella* protozoa, as the results of Example 2. Ginkgolide B and Ginkgolide C showed killing ability against *Eimeria tenella* protozoa at concentrations above 125 ppm. Ginkgolide A showed killing ability against *Eimeria tenella* protozoa at concentrations above 31 ppm. Ginkgolide and Quercus quercetin showed killing ability against *Eimeria tenella* protozoa at concentrations above 63 ppm, and showed a mortality rate of over 50% at concentrations above 500 ppm. Kaempferol showed killing ability against protozoa at concentrations above 125 ppm, and showed a mortality rate of over 50% at concentrations above 500 ppm.

[0203] Example 5. Evaluation of the acid resistance of Ginkgo biloba leaf powder

[0204] In this embodiment, the acid tolerance of Ginkgo biloba leaves was evaluated. Hydrochloric acid (HCl) solution was added to the Ginkgo biloba leaf powder prepared in Examples 1 and 2 above, adjusting the pH to 2.0, 2.5, and 3.0, respectively, and then allowed to stand at 40°C for 1 hour. Then, by adding sodium hydroxide (NaOH) solution to neutralize (pH 7.0), 200,000 Eimeria giant coccidia protozoa (spores) were exposed to different concentrations of Ginkgo biloba leaf powder diluted with PBS solution (10 ppm to 500 ppm) and reacted at 41°C for 4 hours. The protozoan mortality rate (%) was then measured using a method similar to that in Example 2 above, and is shown in Table 13 below. In Table 13, "protozoa alone" refers to the untreated group of Ginkgo biloba leaf powder, and the control group refers to the group treated with Ginkgo biloba leaf powder without hydrochloric acid treatment. As shown in Table 13, it can be confirmed that Ginkgo biloba leaf powder does not lose its killing effect on protozoa that induce coccidiosis even under strongly acidic conditions.

[0205] Table 13

[0206]

[0207] Example 6. Evaluation of the heat resistance of Ginkgo leaf powder

[0208] In this embodiment, the heat resistance of Ginkgo biloba leaves was evaluated. The Ginkgo biloba leaf powder prepared in Examples 1 and 2 above was exposed to 85°C–95°C for 15 minutes, then cooled and diluted with PBS solution to concentrations of 1 ppm, 10 ppm, 50 ppm, and 100 ppm. 200,000 *Eimeria tenella* protozoa were reacted with different concentrations of Ginkgo biloba leaf powder (1 ppm–100 ppm) at 41°C for 4 hours. The protozoan mortality rate (%) was then measured using a method similar to that in Example 2, as shown in Table 14. In Table 14, "protozoa alone" refers to the group untreated with Ginkgo biloba leaf powder, and the control group refers to the group treated with Ginkgo biloba leaf powder without high-temperature treatment.

[0209] As shown in Table 14, it can be confirmed that Ginkgo leaf powder does not lose its killing effect on protozoa that induce coccidiosis even under adverse conditions of high temperature.

[0210] Table 14

[0211]

[0212] From the above description, those skilled in the art will understand that this application can be implemented in other specific forms without altering the technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of this application should be interpreted to include all changes or modifications and equivalent concepts derived from the meaning and scope of the claims described below, and not the detailed description above. sequence list <110> CJ CheilJedang Corporation <120> Anticoccidial compositions containing ginkgo leaves and their uses <130> OPP20211039KR <150> KR 10-2020-0125244 <151> 2020-09-25 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Synthetic E. tenella ITS-1 forward primer <400> 1 tggaggggat tatgagagga 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Synthetic E. tenella ITS-1 reverse primer <400> 2 caagcagcat gtaacggaga 20

Claims

1. The use of at least one compound selected from the group consisting of ginkgolides A, ginkgolides B, ginkgolides C, ginkgolide and their salts as an active ingredient in the preparation of a composition for the prevention, relief or treatment of coccidiosis. in, Ginkgolide A or its salt is contained in the composition at a concentration of 31 ppm or higher; Ginkgolide B or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide C or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide or its salt is contained in the composition at a concentration of 63 ppm or higher; and The coccidiosis mentioned above is caused by Eimeria genus ( Eimeria sp. Protozoa-induced.

2. The use according to claim 1, wherein, The compound also includes at least one selected from the group consisting of quercetin, kaempferol and their salts as an active ingredient.

3. The use according to claim 1, wherein, The prevention, mitigation or treatment of coccidiosis is selected from at least one of the following groups (1) to (4): (1) Reduce at least one of the groups selected from those consisting of lesion score, fecal oocyst excretion, and mortality; (2) Inhibit weight loss caused by coccidiosis; (3) Increase the anticoccidial index (ACI); and (4) Reduce cell invasion of Eimeria protozoa, proliferation of protozoa in cells, or both.

4. The use of at least one compound selected from the group consisting of ginkgolides A, B, C, ginkgolide, and their salts as an active ingredient in the preparation of a pharmaceutical composition for the prevention or treatment of coccidiosis. in, Ginkgolide A or its salt is contained in the composition at a concentration of 31 ppm or higher; Ginkgolide B or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide C or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide or its salts are contained in the composition at a concentration of 63 ppm or higher, and The coccidiosis mentioned above is caused by Eimeria genus ( Eimeria sp. Protozoa-induced.

5. The use according to claim 4, wherein, The compound also includes at least one selected from the group consisting of quercetin, kaempferol and their salts as an active ingredient.

6. The use of at least one compound selected from the group consisting of ginkgolides A, B, C, ginkgolide, and their salts as an active ingredient in the preparation of an antiprotozoan composition against Eimeria protozoa. in, Ginkgolide A or its salt is contained in the composition at a concentration of 31 ppm or higher; Ginkgolide B or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide C or its salt is contained in the composition at a concentration of 125 ppm or higher; Ginkgolide or its salts are contained in the composition at a concentration of 63 ppm or higher.

7. The use according to claim 6, wherein, The compound also includes at least one selected from the group consisting of quercetin, kaempferol and their salts as an active ingredient.

Images