KR20150041698A - Pharmaceutical composition comprising genipin and crocin as effective components for prevention or treatment of diseases related to thrombosis and health functional food comprising the same - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and functional health food for preventing or treating thrombotic diseases, containing genipin and crocin exhibiting strong platelet aggregation inhibitory activity as active ingredients. A mixture containing genipin and crocin as active ingredients of the pharmaceutical composition and functional health food for preventing or treating thrombotic diseases of the present invention strongly inhibits human platelet aggregation, thereby efficiently suppressing thrombosis.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition and a health functional food for preventing or treating thrombotic diseases containing jenipine and crocin as an effective ingredient exhibiting a synergistic effect of inhibiting platelet aggregation, THROMBOSIS AND HEALTH FUNCTIONAL FOOD COMPRISING THE SAME}

The present invention relates to a pharmaceutical composition and a health functional food for the prevention or treatment of a thrombotic disease which contains, as an active ingredient, genipin and crocin which exhibit strong human platelet aggregation inhibiting activity.

As a constituent of human body, blood has various important functions such as oxygen and nutrients, the function and buffering function of waste products, maintenance of body temperature, control of osmotic pressure and maintenance of ion balance, maintenance of moisture, regulation of fluidity, maintenance and regulation of blood pressure, have. Therefore, blood and blood circulation are essential for the survival of human beings. In case of damage of blood vessels, blood clots should be rapidly generated to prevent blood loss.

However, excessive blood clot formation in the blood vessels interferes with blood circulation, and in severe cases, blood vessels are blocked to cause serious damage to cardiovascular system and brain tissue, resulting in thrombosis. Such thrombosis is caused by the following: 1) abnormal vascular endothelial cells, 2) excessive coagulation activity of blood coagulation factors (XII factor, XI factor, IX factor, X factor etc.) and blood coagulation enzyme (thrombin, prothrombin enzyme) 4) The abnormal function of the thrombus dissolution system is known to be responsible for these causes. These causes are caused individually or in combination, resulting in excessive blood clot formation in the blood vessels.

For the prevention and treatment of thrombotic diseases, currently 1) various anticoagulants such as heparin and coumarin which inhibit the activity of blood coagulation factors to inhibit thrombogenesis, 2) anti-platelet aggregation inhibitors such as aspirin Thrombocytopenia, and 3) a thrombolytic agent such as europaea, which inhibits thrombus formation by dissolving already generated thrombus. However, they are not only very expensive, but also have limited use due to hemorrhagic side effects, gastrointestinal disorders, and hypersensitivity reactions.

KR 10-2008-0002770 A

 Zhang, et al., 2013. Antitrombotic activities of aqueous extracts from Gadenia jasminoides and its main constituents. Pharmacology Biology, 51: 221-225   Suzuki, et al., 2001. Antithrombotic effect of geniposide and genipin in the mouse thrombosis model. Plant Med. 67: 807-810   Thushara et al., 2013. Crocin, a dietary additive protects platelets from oxidative stress induced apoptosis and inhibit platelet aggregation. Mol. Cell Biochem 373: 73-83

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in order to solve the problems of the prior art as described above, and it is an object of the present invention to provide a method for inhibiting platelet aggregation, A pharmaceutical composition for preventing or treating thrombotic diseases, and a health functional food.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating thrombotic diseases comprising genipin represented by the following formula 1 and crocin represented by the following formula 2 as an active ingredient Gt;

[Chemical Formula 1]

(2)

In addition, the pharmaceutical composition of the present invention may further comprise aspirin, which is an antithrombotic agent commercially available as an active ingredient.

The thrombotic disease is caused by acute myocardial infarction, chest pain, dyspnea, loss of consciousness, ischemic stroke, hemorrhagic stroke, deep vein thrombosis, lower limb edema, pain, acute peripheral arterial atresia, deep vein thrombosis, Thrombosis, acute renal vein occlusion, cerebral sinus thrombosis and central retinal vein occlusion.

The present invention also provides a health functional food for preventing or improving thrombotic diseases containing genipin and crocin described above.

The pharmaceutical composition for the prevention or treatment of thrombotic diseases of the present invention and the mixture comprising genipin and crocin as an active ingredient of the health functional food strongly inhibit the aggregation of human platelets, There is an effect that can be suppressed.

FIG. 1 is a graph showing the activity of human platelet aggregation inhibitory activity of a 1: 1 equivalent mixture of genipin, crocin, genipin and crocin. (25 μg / ml), crocin (25 μg / ml), and aspirin (0.25 mg / ml: 1400 μM) as a solvent control from the upper left, DMSO, aspirin , genipin (25 μg / ml) + crocin (25 μg / ml), respectively.

Hereinafter, the present invention will be described in detail.

The present invention relates to a pharmaceutical composition comprising genipin, methyl (1S, 2R, 6S) -2-hydroxy-9- (hydroxymethyl) -3-oxabicyclo [4,3,0] nona-4,8-diene -5-carboxylate and crocin (crocetin digentiobiose ester) represented by the following formula (2) as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating a thrombotic disease.

[Chemical Formula 1]

(2)

Genipin and crocin exhibit weak human platelet aggregation inhibitory activity in each case. However, these mixtures exhibit very strong platelet aggregation inhibitory activity and are useful as pharmaceuticals for the prevention or treatment of thrombotic diseases Can be used as a composition.

The genipin and crocin can be obtained commercially (Sigam, USA) or isolated from medicinal plants such as chemical synthesis or gardenia.

Jenny pin of the present invention has recently been shown to inhibit platelet aggregation and has been reported to inhibit thrombogenesis by 67.6% when administered at a dose of 20 mg / kg (Zhang, et al., 2013. Pharmacology Biology, 51: 221-225).

In addition, it has been reported that crocin of the present invention can efficiently inhibit collagen-induced platelet aggregation on a test tube scale as a recently discovered platelet aggregation inhibiting function (Thushara et al., 2013. Mol. Cell Biochem 373: 73-83).

However, the mechanism of antiplatelet function of zenithpin and crocin has not been known to date, and it is not known that the mixture of zeniprin and crocin exhibits synergistic effects of platelet aggregation inhibition function. Therefore, the mixture of the above-mentioned jenny pin and crocin exhibits a strong synergistic effect than the effect of platelet aggregation inhibition indicated when using only jenny pin or crocin, so that the mixture can be used for the prevention or treatment of various diseases related to the thrombotic disease .

In particular, a recent paper (78%) reported a mortality rate of 78% in a rodent experiment with orally administered 200 mg / kg of jennifin (Hou et al., 2008. Metabolism and pharmacokinetics of genipin and geniposide in rats, Food and Chemical Toxicology, 46: 2764-2769), it is important to use a minimum amount of Jenny pin. Therefore, in order to prevent and treat thrombotic diseases without severe side effects such as lethality, a mixture of Jenny pin and cloche is used as an antithrombotic agent Can reduce the amount of usage due to the synergy effect.

The mixing ratio of the jenny-pin and crocine can be adjusted in the range of 1: 0.1 to 10, preferably 1: 0.5 to 5, and most preferably 1: 1.

In addition, the present invention may further include aspirin (acetylsalicylic acid) in addition to zeniprin and crocin as an active ingredient of the pharmaceutical composition of the present invention.

When genipin and crocin are used in combination, they exhibit very strong platelet aggregation inhibitory activity. However, synergistic effect is further enhanced when aspirin is additionally used.

The mixing ratio of the Jenny pin, crocin and aspirin can be adjusted in the range of 1: 0.1-10: 10-100, preferably 1: 0.5-5: 5 - 50, most preferably 1: 20 by weight.

Such thrombotic diseases include, for example, arterial thrombosis such as acute myocardial infarction, chest pain, dyspnea, loss of consciousness, ischemic stroke, hemorrhagic stroke, headache, dyskinesia, sensory abnormalities, personality changes, Deep vein thrombosis, acute renal vein thrombosis, cerebral sinus thrombosis, central retinal vein occlusion, and the like can be cited as examples of venous thrombosis such as pulmonary thrombosis, deep vein thrombosis, deep vein thrombosis, lower limb swelling, pain and acute peripheral artery occlusion. have.

The pharmaceutical composition comprising the mixture of the present invention and the clenidine may be formulated into oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, sterilized Injections of injection solutions, and the like, and they can be administered through various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, and the like.

Such pharmaceutical compositions may further comprise carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, But are not limited to, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

In a preferred embodiment, the solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, for example starch, calcium carbonate, Sucrose, lactose, gelatin and the like are mixed and formulated. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used.

Examples of the oral liquid preparation include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, Perfumes, preservatives, and the like.

As a preferable specific example, the preparation for parenteral administration includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-drying agents, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, and the like.

The mixture comprising the present zenith fins and the crocin is administered in a pharmaceutically effective amount. In the present invention, a "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level will depend on the type of disease, severity, The sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

In a preferred embodiment, the effective amount of the mixture of jenipine and clocin may vary depending on the age, sex, and body weight of the patient, and is generally in the range of 1 to 10 mg per kilogram body weight (in this case, 0.5 to 5 mg of jenipine, , Preferably 2 to 5 mg per day or every other day, or one to three divided doses per day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like.

Further, the present invention provides a health functional food for preventing or ameliorating a thrombotic disease, which comprises a mixture of jenny pin and crocin as an active ingredient.

The mixing ratio of the jenny-pin and crocine can be adjusted in the range of 1: 0.1 to 10, preferably 1: 0.5 to 5, and most preferably 1: 1.

The health functional food of the present invention can be variously used for foods and beverages effective for preventing or improving thrombotic diseases. Examples of the foods containing the mixture of jenny pin and crocin of the present invention include various foods, beverages, gums, tea, vitamin complex, health supplement foods and the like, and may be in the form of powder, granule, tablet, capsule or beverage .

The mixture of the present invention jenipine and croscreen may generally be added in an amount of 0.01 to 0.2% by weight of the total food, and the health beverage composition may be added in an amount of 0.005 to 0.5 g based on 100 ml.

The health functional food of the present invention may contain, as an additional ingredient, a food-acceptable food-aid additive such as natural carbohydrates and various flavors, in addition to containing the above-mentioned compound as an essential ingredient in the indicated ratio.

Examples of the natural carbohydrate include sugar sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol.

As the flavor, natural flavoring agents such as tautatin and stevia such as rebaudioside A or glycyrrhizin, and synthetic flavorings such as saccharin and aspartame may be used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and heavy stabilizers, pectic acid and its salts, alginic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for producing natural fruit juice, fruit juice drink, vegetable drink and the like. These components may be used independently or in combination. The ratio of such additive is generally selected in the range of 0.01 to about 5 parts by weight per 100 parts by weight of the mixture of the jenny pin and crocine of the present invention.

Hereinafter, the specific method of the present invention will be described in more detail by way of examples. The following examples are only a preferred embodiment of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[ Example ]

Example  One: Jenny Finn , Crocin  And Jenny Finn Crocin  Mixture of Evaluation of blood clotting enzyme / blood coagulation factor inhibitory activity

Jenny pin and crocin (Sigma, USA) having a purity of 99% or more were purchased. Each of these and mixtures thereof was evaluated for blood clotting inhibitory activity.

Blood clotting enzyme inhibition was measured by thrombin time and prothrombin time, and the inhibitory activity of blood clotting factors by endogenous thrombus formation was measured by apitime time (Sohn et al ., 2004. Kor J. Pharmacogn 35, 52-61; Kwon meat al ., 2004. J. Life Science , 14: 509-513; Li et al. 2010. J. Life Science , 20, 922-928). The specific measurement method is as follows.

Thrombin  time( Thrombin Time )

50 [mu] l 0.5 U thrombin (Sigma Co., USA) and 20 mM CaCl ₂ (Control plasma, Thermo Fisher Scientific Inc, MA, USA) was added to the tube of Amelung coagulometer KC-1A (Japan) for 2 minutes. After the addition, the time until the plasma coagulated was measured. As a control, aspirin (Sigma Co., USA) was used and DMSO was used as a solvent control instead of the sample. DMSO showed a clotting time of 32.1 sec, and the results were expressed as the mean value of the experiments repeated three or more times.

Prothrombin  time( prothrombin time )

Add 70 μl of standard plasma (MD Pacific Co., China) and appropriate concentrations of genepin, crocin or a mixture thereof to the tube of Amelung coagulometer KC-1A (Japan), incubate at 37 ° C for 3 minutes, add 130 μl of PT reagent (Control plasma, Thermo Fisher Scientific Inc, MA, USA) was used as the average value of the experiment in which the time until coagulation was repeated three times. As a control, aspirin (Sigma Co., USA) was used and DMSO was used as a solvent control instead of the sample. DMSO showed a clotting time of 18.1 sec, and the results were expressed as the mean value of the experiments repeated three or more times.

aPTT  ( activated Partial Thromboplastin Time )

100 μl of plasma (control plasma, Thermo Fisher Scientific Inc, MA, USA) and appropriate concentrations of genepin, crocin or a mixture thereof were added to tubes of Amelung coagulometer KC-1A (Japan) , 50 μl of aPTT reagent (Sigma, ALEXIN ^™ , USA) was added and incubated at 37 ° C. for 3 minutes. After the addition of 50 μl CaCl ₂ (35 mM), the time until the plasma coagulated was measured. As the solvent control, DMSO was used instead of the sample. In this case, the solidification time of 55.1 seconds was shown, and the results were shown as the average value of three repeated experiments.

According to the above measurement method, the blood coagulation inhibitory activity of a mixture of jennypin, crocin, and jennifin and crocin was evaluated at 0.5 mg / ml, 1.5 mg / ml, and 2.5 mg / ml concentration and no significant activity was observed , And thus it was concluded that there is no blood coagulation inhibitory activity of jenipine, crocin and their equivalent mixture (no result).

Example  2: Jenny Finn , Crocin  And Jenny Finn Crocin  Mixture of Evaluation of Platelet Aggregation Inhibitory Activity

The platelet aggregation inhibitory activity of each of the jenny fins and the crocins and the mixture thereof in Example 1 was evaluated in accordance with the following method. As a control, aspirin (Sigma Co., USA) was used and DMSO was used as a solvent control instead of the sample.

Platelet Aggregation Inhibitory Activity Platelet aggregation inhibition activity )

Platelets were mixed with 3.2% sodium citrate solution in a ratio of 1: 9 from healthy human whole blood, and centrifuged at 1,100 rpm for 10 minutes to obtain upper layer platelet rich plasma (PRP). The platelets were washed with a washing buffer (138 mM NaCl, 2.7 mM KCl, 12 mM NaHCO ₃ , 0.36 mM NaH ₂ PO ₄ , 5.5 mM Glucose, 1 mM EDTA, pH 6.5). Thereafter, the cells were resuspended in a suspending buffer (138 mM NaCl, 2.7 mM KCl, 12 mM NaHCO ₃ , 0.36 mM NaH ₂ PO ₄ , 5.5 mM Glucose, 0.49 mM MgCl ₂ , 025% gelatin, pH 7.4) and incubated at 3,000 rpm for 10 minutes After centrifugation, the cells were resuspended in a suspending buffer and the platelet count was adjusted to ⁴ × ^{10 9} / ml. Then platelet aggregation was measured at 37 ° C using a whole-blood agarometer (Chrono-log, USA) after 2.5 μl of collagen was added to 1 ml of suspension and reacted for 5 minutes.

Platelet aggregation inhibitory activity of jennifin was 80.1% at the concentration of 25 μg / ml (19.9% inhibition of platelet aggregation), and 48.7% of the platelet aggregation ability at 50 μg / ml Indicating good platelet aggregation inhibitory activity. On the other hand, crocin exhibited weak platelet aggregation inhibition ability at a concentration of 25 μg / ml, showing 82.7% platelet aggregation ability (17.3% platelet aggregation inhibition ability), and 76.2% platelet aggregation ability at 50 μg / ml It stopped showing. Platelet aggregation was 91% and 101.6%, respectively, when platelets were treated at a concentration of 12.5 μg / ml, respectively (Table 1, Fig. 1).

However, the same amount of mixture of zenithpin and crocin exhibited more synergistic inhibition of platelet aggregation than the respective substances. First, when the concentrations of jennypin and clocin were respectively 12.5 μg / ml, the aggregation rate decreased to 76.3%, indicating that the aggregation inhibition was better than that of 25 μg / ml or 25 μg / ml of jenny pin, In the case of mixture of 25 μg / ml each of pin and crocin, only 23.6% platelet aggregation (76.4% platelet aggregation inhibition) was observed. It was found that the synergistic effect of platelet aggregation inhibition was very strong when considering the platelet aggregation ability of 48.7% at the concentration of 50 μg / ml of JENNIFINE and the platelet aggregation ability of 76.2% at the treatment of 50 μg / mL of crocin (Table 1 , Fig. 1). Actually, when 50 μg / ml of each was mixed, the platelet aggregation inhibition rate was almost 100% because there was almost no platelet aggregation at 0.2%. This may be an indication of the platelet aggregation inhibitory effect of the zeinifine and the crocin mixture, considering that aspirin (product name: Protect, Bayer), which is used as a platelet aggregation inhibitor, has a platelet aggregation of 37.5% at a concentration of 500 μg / ml.

Sample / Control burglar
(Amplitude
: ohm) inclination
(Slope) Extension time
(Lag time: seconds) ^* Falling area
(Area under) Cohesion rate
(%) DMSO (DMSO) 17 3 31 123.6 100.0 Aspirin (500 μg / mL, 2,800 μM) 8 One 41 46.4 37.5 Aspirin (250 μg / mL, 1,400 μM) 16 2 25 102.5 82.9 Jenipine (50 μg / mL) 8 One 40 60.2 48.7 Jenipine (25 μg / mL) 14 2 26 99.0 80.1 Jenipine (12.5 μg / mL) 17 3 32 112.5 91.0 (50 < RTI ID = 0.0 > pg / mL) 14 2 26 94.2 76.2 Croscreen (25 μg / mL) 14 2 28 102.2 82.7 (12.5 μg / mL) 17 3 28 125.7 101.6 Jenny pin (50 μg / mL) +
(50 < RTI ID = 0.0 > pg / mL) 0 0 58 0.3 0.2 Jenny pin (25 μg / mL) +
Croscreen (25 μg / mL) 4 One 58 29.2 23.6 Jenny pin (12.5 μg / mL) +
(12.5 μg / mL) 13 2 32 94.4 76.3

* Smaller falling area means no platelet aggregation

On the other hand, it is also possible to use geniposide (1S, 4αS, 7αS) -1- (β-D-Glucopyranosyoxy) 1,4a, 5,7a-tetrahydro-7- (hydroxymethyl) cyclopenta [c] pyran -4-carboxylic acid methyl ester is a glycoside conjugated to a single glucose molecule in the genitin (Zhang, et al., 2013. Pharmacology Biology, 51: 221-225) Showed a platelet aggregation inhibitory effect which was similar to that of jennifin at a dose of 5 times higher than that of jennifin (Table 2). In addition, the synergistic effect of the mixture of xenon-foside and crocin was not significant (Table 2), confirming that the synergistic effect of the use of the mixture of xenon and crocin was specific.

(3)

Sample / Control burglar
(Amplitude
: ohm) inclination
(Slope) Extension time
(Lag time: seconds) ^* Falling area
(Area under) Cohesion rate
(%) DMSO (DMSO) 19 3 23 134.6 100.0 Aspirin (500 μg / mL, 2,800 μM) 5 One 10 37.6 27.9 Aspirin (250 μg / mL, 1,400 μM) 12 2 16 68.0 50.5 Jenny foside (250 μg / mL) 11 2 38 78.3 58.1 JENNY POID (25 μg / mL) 21 3 20 143.9 106.9 JENNY POID (25 μg / mL)
+ Crocin (25 [mu] g / mL) 14 2 22 104.5 77.6

* Smaller falling area means no platelet aggregation

Example  3: Jenny Finn Crocin  Evaluation of synergistic effect of aspirin on platelet aggregation inhibitory activity of a mixture

Synergistic effects were observed when an aspirin (commercial platelet aggregation inhibiting antithrombotic agent) was further mixed in the same amount mixture of xanthine and crocin.

First, as shown in Table 3, when 25 μg / ml of aspirin and 250 μg / ml of aspirin were mixed, the change of the platelet aggregation rate was within 1%, indicating 80.1 ~ 81.1% of aggregation rate. In addition, when 250 μg / ml of aspirin was added to 25 μg / ml of crocin, the change of platelet aggregation rate was also within 1.7%, indicating 81.0 ~ 82.7%.

However, the addition of aspirin at a concentration of 250 μg / ml in addition to the mixture of jennypin and crocin (each concentration of 12.5 μg / ml: total of 25 μg / ml) decreased the agglutination rate from 76.3% to 29.7% Showed very strong synergistic inhibition of platelet aggregation. Although the exact mechanism is not known, the synergistic effect of the mixture of JENNIFINE and CROSINE on existing aspirin patients or the general public indicates that it is possible to further reduce the use of JENNIFINE and CROSINE, Suggesting that the use of aspirin may be associated with side effects. Therefore, this result suggests that Jenipin, Crocine, and Aspirin all have different targets in the final inhibition mechanism of platelet aggregation.

Sample / Control burglar
(Amplitude
: ohm) inclination
(Slope) Extension time
(Lag time: seconds) ^* Falling area
(Area under) Cohesion rate
(%) DMSO (DMSO) 17 3 31 123.6 100.0 Aspirin (500 μg / mL, 2,800 μM) 8 One 41 46.4 37.5 Aspirin (250 μg / mL, 1,400 μM) 16 2 25 102.5 82.9 Jenipine (25 μg / mL) 14 2 26 99.0 80.1 Jenny pin (25 μg / mL) +
Aspirin (250 μg / mL) 16 2 32 100.3 81.1 Croscreen (25 μg / mL) 14 2 28 102.2 82.7 Crocin (25 μg / mL) +
Aspirin (250 μg / mL) 16 2 36 100.2 81.0 Jenny pin (12.5 μg / mL) +
(12.5 μg / mL) 13 2 32 94.4 76.3 Jenny pin (12.5 μg / mL) +
(12.5 μg / mL) +
Aspirin (250 μg / mL) 5 One 33 36.8 29.7

* Smaller falling area means no platelet aggregation

Example  4: Jenny Finn , Crocin  And Jenny Finn Crocin  Human erythrocyte hemolytic activity of the mixture

As a result of measuring the hemolytic activity of human erythrocytes in the jenny pin and the crocin mixture of Example 1, human hemolytic hemolytic activity did not show up to 200 μg / ml concentration as in the case of each of jennifin and crocin (Table 4). Thus, the use of a mixture of jennifin and crocin showed no additional acute toxicity.

sample Human hemolytic hemolytic activity (%) Jenny pin (100 μg / ml) 5.4 ± 0.24 (100 μg / ml) 1.6 ± 0.51 (100 mu g / ml) + crocin (100 mu g / ml) mixture 5.0 ± 0.79 Triton X-100 (0.1% 100.0 ± 1.20 Amphotericin B 62 μg / ml 97.0 3.55

Claims (4)

A pharmaceutical composition for preventing or treating a thrombotic disease comprising genipin represented by the following formula (1) and crocin represented by the following formula (2) as an active ingredient.
[Chemical Formula 1]

(2)

The pharmaceutical composition according to claim 1, wherein the active ingredient further comprises aspirin. 4. The method of claim 1 or 2, wherein the thrombotic disease is selected from the group consisting of acute myocardial infarction, chest pain, dyspnea, loss of consciousness, ischemic stroke, hemorrhagic stroke, deep vein thrombosis, Acute peripheral arterial occlusion, deep vein thrombosis, portal vein thrombosis, acute renal vein occlusion, cerebral sinus thrombosis, and central retinal vein occlusion. A health functional food for preventing or ameliorating a thrombotic disease comprising genipin and crocin according to claim 1.

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