KR102085115B1 - Pharmaceutical composition for preventing or treating hematoposis disorder diseases comprising maghemite-saponins nanoparticles - Google Patents

Abstract

The present invention relates to a composition for treating hematopoietic dysfunction comprising a maghemite-saponin nanoconjugate as an active ingredient, wherein the maghemite-saponin nanoconjugate improves red blood cell reduction in anemia due to iron deficiency, chemicals or The maghemite-saponin nanoconjugates are associated with hematopoietic dysfunction, confirming that the number of erythrocytes, leukocytes, platelets, or neutrophil granulocytes in animal models showing hematopoietic deterioration such as hematopoiesis decrease by radiation treatment is restored to normal levels. It may be provided as a composition for preventing or treating a disease, and may be combined with a chemical or radiation therapy.

Description

Pharmaceutical composition for preventing or treating hematoposis disorder diseases comprising maghemite-saponins nanoparticles}

The present invention relates to a composition for treating hematopoietic dysfunction comprising a maghemite-saponin nanoconjugate as an active ingredient.

The hematopoietic system of the human body is inherently or very sensitive to exposure to various harmful factors such as radiation or some chemotherapeutic drugs, resulting in related diseases resulting from decreased hematopoietic function, ie, reduced blood cells such as red blood cells, white blood cells and platelets.

Representative diseases due to erythrocyte abnormality are anemia, and "anemia" is a condition in which the number of erythrocytes and hemoglobin in the blood is lower than the normal value according to the measurement of the specific volume of blood, and according to the cause of aplastic anemia, hemolytic anemia, Iron deficiency anemia and sickle cell anemia.

Aplastic anemia exhibits red blood cell regeneration deficiency and resistance to treatment, and hemolytic anemia is anemia caused by abnormal hemolysis of red blood cells. Iron deficiency anemia is anemia most commonly found in medical outpatient practice. Iron is an essential microorganism for heme production, and iron deficiency leads to decreased heme production, which is hemoglobin. It causes the production of, resulting in microcytic anemia in which the size of red blood cells is reduced.

In order to treat such iron-deficiency anemia, oral iron, parenteral iron, intravenous injection, blood transfusion, and the like are used. Among them, the frequency of the use of oral iron is particularly high. Oral iron is largely divided into non-heme iron and heme iron. Currently non-heme iron formulations in the domestic market include ferrous sulfate, polysaccharide iron complex, iron protein succinylate, ferric hydroxide poly maltose, and carbohydrate. Carbonyl iron is a disadvantage that is often accompanied by gastrointestinal disorders such as bloating, abdominal pain, constipation and diarrhea. Heme iron is a problem that the side effects of the above-mentioned non-heme iron is low, but the iron content is low, the rapidity as a therapeutic agent is poor.

Intravenous injection, a parenteral iron product, must satisfy the conditions that iron should be readily available for hemoglobin synthesis, have no local or systemic side effects, and have sufficient storage stability due to half-life. Old iron powders include iron-dextran (e.g. InFed, Dexferrum), iron-gluconate complexes (Ferrlecit) and iron-sucrose (Venoferrum). .

Iron-dextran is the first parenteral iron product on the market in the United States, with a high incidence of anaphylactoid reactions (respiratory distress, asthma cough, chest pain, hypotension, urticaria, hypersensitivity reactions such as angioedema) Parenteral iron products (eg iron-sucrose and iron-gluconate) do not contain dextran and have a low frequency of anaphylaxis, but these compounds are toxic to iron compounds when administered and have high pH, high molarity, etc. There is a problem in that the dosage and the rate of administration are limited according to the physical properties of the.

Recently, ferumoxytol (ferumoxytol) based on iron oxide nanoparticles has been developed as an iron-containing preparation for parenteral administration rather than iron-complex. Ferrumoxytol is more stable than other parenteral iron products, such as iron-dextran, iron-sucrose, and iron-gluconate, but has a low concentration of free iron and should be administered twice a week to administer 1 g of iron. And, accordingly, there is a problem that causes the discomfort of the patient due to the injection cost and the tube and the injection solution.

In addition, chemotherapy with radiation or chemotherapy is a leading cause of bone marrow hematopoietic function, such as blood cell reduction. To date, chemotherapy with radiation and chemotherapy with radiation are used in addition to specific cytotoxicity to target tumor cells. Nonspecific cytotoxicity to other body cells is a problem, and in particular, bone marrow hematopoietic function, which requires active cell division and differentiation, can be suppressed under the influence of chemotherapy. Bone marrow suppression is a decrease in hematopoietic function in the bone marrow, resulting in a decrease in bone marrow proliferation, decreased blood cell counts, decreased peripheral blood leukocytes, especially neutrophil granulocytes, decreased or increased platelets, and aplastic anemia. It can fall and even be life threatened.

As a treatment used to overcome the bone marrow suppression, which is a side effect of the anti-cancer treatment, there are methods for administering various growth factors. Typically, granulocyte macrophage colony stimulating factor (rhGM-CSF), granulocyte colony stimulating factor (rhG-CSF), etc. There is a cure used. These drugs have significant effects on increasing leukocytes during chemotherapy, but they are expensive and difficult to apply to most patients, are difficult to use with chemotherapy, they cannot be used for prophylaxis and only when white blood cells are reduced. It can be used, or it will cause toxic side effects. In addition, gene therapy is still in the early stages of development. Ego bone marrow transplantation is used in parallel with mass chemotherapy but has a problem that is difficult to apply repeatedly.

Therefore, in order to prevent or treat anemia due to hematopoietic deterioration and blood cell reduction, which are side effects accompanying cancer chemotherapy and radiation therapy, it is necessary to develop safer, more efficient and economical therapeutic agents.

Korean Patent Publication No. 2013-0113575 (published Oct. 16, 2013)

The present invention to prevent or treat hematopoietic dysfunction associated with blood cell reduction and hematopoietic dysfunction due to bone marrow suppression, which is a side effect of cancer chemotherapy or radiation therapy, as the active ingredient It is to provide a composition containing.

The present invention can provide a pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient.

The present invention can provide a health food for preventing or improving hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient.

In addition, the present invention contains a maghemite-saponin nanoconjugate as an active ingredient, the nanoconjugate prevents or improves the hematopoietic dysfunction to prevent or improve the number of red blood cells, leukocytes, platelets or neutrophil granulocytes by chemical or radiotherapy Prophylactic or improvement aids may be provided.

According to the present invention, maghemite-saponin nanoconjugates improve erythrocyte reduction in anemia due to iron deficiency, and erythrocytes, leukocytes, platelets or erythrocytes of animal models in which hematopoietic function decreases such as blood cell reduction by chemical or radiation treatment. As it is confirmed that the number of neutrophil granulocytes is restored to the normal level, the maghemite-saponin nanoconjugate may be provided as a composition for preventing or treating hematopoietic dysfunction disorder, and may be combined with chemical or radiation therapy. have.

1 is a result of confirming the effect of improving the levels of erythrocytes and hemoglobin after oral administration of the maghemite-saponin nanoconjugate for 4 weeks in an animal model induced iron deficiency anemia. * p <0.05, ** p <0.01 and *** p <0.001 vs. control
Figure 2 shows the effect of improving the levels of leukocytes, platelets and neutrophil granulocytes after oral administration of maghemite-saponin nanoconjugates to animal models induced hematopoietic dysfunction by intraperitoneal administration of cyclophosphamide (CYP) for 4 weeks. to be. * p <0.05, ** p <0.01 and *** p <0.001 vs. control
Figure 3 is a result of confirming the effect of improving the levels of leukocytes, platelets and neutrophil granulocytes after oral administration of maghemite-saponin nanoconjugate for 4 weeks in an animal model induced hematopoietic dysfunction by radiation treatment. * p <0.05, ** p <0.01 and *** p <0.001 vs. control

The present invention can provide a pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient.

The nanoconjugate may be directly bonded to the saponin on the surface of maghemite, the saponin may be a plant-derived dammarene-based saponin.

The dammarene-based saponins may be selected from the group consisting of ginseng-derived ginsenosides and dodol-derived fenenosides.

In more detail, the ginseng-derived ginsenoside may be selected from the group consisting of ginsenoside Rg3, ginsenoside F2, and ginsenoside CK, and the dodol-derived fenenoside may be zipenoside XVII, but is not limited thereto. It doesn't work.

The maghemite-saponin nanoconjugates can increase the number of red blood cells, white blood cells, platelets or neutrophil granulocytes.

The hematopoietic dysfunction disorder may be selected from the group consisting of anemia, malignant lymphoma, leukemia, myeloid thrombocytopenia, idiopathic thrombocytopenic purpura, thrombocytopenia, lymphocytosis, monocytopenia, granulocytopenia and myelodysplastic syndrome.

The anemia may be selected from the group consisting of aplastic anemia, hemolytic anemia, iron deficiency anemia and sickle cell anemia, but is not limited thereto.

The maghemite of the present invention is a magnetic iron oxide nanoparticles, iron oxide widely present in nature is a material easily synthesized even in the laboratory environment to produce magnetic iron oxide particles having a special magnetic properties when making the iron oxide nanoscale, Due to the characteristics of biochemical, magnetic, and catalyst, it is being used as a material for drug delivery, magnetic resonance imaging, thermotherapy, and biosensors.

Among the magnetic iron oxide nanoparticles, hematite, magnetite, and maghemite are mainly used. In particular, magnetite and maghemite have strong magnetic properties, and thus are highly active in bio, pharmaceutical, and medical device fields.

However, in order to actually utilize magnetite and maghemite, there is a problem of physicochemical properties that are easily oxidized in the air or in an aqueous solution, and requires a separate technology for blocking and dispersing well into nanoparticles. The pharmacological activity of only a few has been reported, including antimicrobial effects.

The present invention relates to a composition containing nanoparticles of 10 to 100 nm average diameter as an active ingredient by directly binding saponin to maghemite (γ-Fe ₂ O ₃ ), which is a magnetic iron oxide nanoparticle, and an embodiment of the present invention. According to the present invention, the iron-deficient diet was ingested with iron-deficient anemia-induced oral administration of maghemite-saponin nanoconjugates for 4 weeks, and the effect of maghemite-saponin nanoconjugates on erythrocyte and hemoglobin levels was reduced. As a result, as shown in FIG. 1, the anemia index of the experimental groups ingested the maghemite-saponin nanoconjugates was recovered to a normal level, and the maghemite-saponin nanoconjugates were treated with maghemite or saponins alone. It showed more improved efficacy than when, it was confirmed that the equivalent or more effect compared to the comparison group ferritin.

In one embodiment of the present invention, the pharmaceutical composition containing the maghemite-saponin nanoconjugates as an active ingredient may be prepared by injection, granule, powder, tablet, pill, capsule, suppository, gel, suspension, Any formulation selected from the group consisting of emulsions, drops or solutions can be used.

In another embodiment of the present invention, pharmaceutical compositions containing maghemite-saponin nanoconjugates as active ingredients are selected from suitable carriers, excipients, disintegrants, sweeteners, coatings, swelling agents, lubricants, commonly used in the preparation of pharmaceutical compositions, It may further comprise one or more additives selected from the group consisting of glidants, flavors, antioxidants, buffers, bacteriostatics, diluents, dispersants, surfactants, binders and lubricants.

Specifically, carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil can be used, and solid preparations for oral administration include tablets, pills, powders, granules, capsules. And the like, and such solid preparations may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, and the like in the composition. In addition to simple excipients, lubricants such as magnesium styrate and talc may also be used. Oral liquid preparations include suspensions, solvents, emulsions, syrups, and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As a base material of suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

According to one embodiment of the invention the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, nasal, inhaled, topical, rectal, oral, intraocular or intradermal Via the route can be administered to the subject in a conventional manner.

The preferred dosage of the maghemite-saponin nanoconjugate may vary depending on the condition and weight of the subject, the type and extent of the disease, the drug form, the route of administration and the duration, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, but not limited thereto, the daily dosage may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg. Administration may be administered once a day or divided into several times, thereby not limiting the scope of the invention.

In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.

In addition, the present invention can provide a health food for preventing or improving hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient.

The health food may be used with other foods or food additives in addition to the maghemite-saponin nanoconjugates and may be appropriately used according to conventional methods. The mixed amount of the active ingredient may be used for the purpose of its use, for example, for prevention, health or therapeutic It may be appropriately determined depending on the treatment.

The effective dose of the compound contained in the health food may be used in accordance with the effective dose of the therapeutic agent, but may be less than the above range in the case of prolonged intake for the purpose of health and hygiene or health control. It is evident that the component can be used in an amount above the range because there is no problem in terms of safety.

There is no particular limitation on the kind of the health food, for example, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, Drinks, alcoholic drinks, vitamin complexes, etc. are mentioned.

In addition, the present invention contains a maghemite-saponin nanoconjugate as an active ingredient, the nanoconjugate prevents or improves the hematopoietic dysfunction to prevent or improve the number of red blood cells, leukocytes, platelets or neutrophil granulocytes by chemical or radiotherapy Prophylactic or improvement aids may be provided.

The nanoconjugate may be directly bonded to the saponin on the surface of maghemite, the saponin may be a plant-derived dammarene-based saponin.

The dammarene-based saponins may be selected from the group consisting of ginseng-derived ginsenosides and dodol-derived fenenosides.

In more detail, the ginseng-derived ginsenoside may be selected from the group consisting of ginsenoside Rg3, ginsenoside F2, and ginsenoside CK, and the dodol-derived fenenoside may be zipenoside XVII, but is not limited thereto. It doesn't work.

In addition, the chemical agent may be selected from the group consisting of alkylating agent-based drugs, anti-metabolic drugs, antibiotic-based drugs, natural-derived drugs and hormone-based drugs, the adjuvant may be administered in combination with chemicals or radiation.

More specifically, the alkylating agent-based drugs are nitrogen mustards, cyclophosphamide, thiotepa, lomustine, myleran, dacarbazine, and procarbage. It can be selected from the group consisting of Procarbazine, the anti-metabolic drug is Fluorouracil (5-FU), Tegafur (FT-207), Tegadifur (FD) -1), Compound Tegafur (UFT), Futron (5-DFUR), Methotrexate (MTX), Aminopterin, Cytosine arabinoside (Ara-c), Cyclocytidine, Cyclocytidine hydrochloride, Hydroxycarbamide (HU), Inosine dialdehyde, Adenosine dialdehyde ), Guanazole and 6-mercaptopurine (6-Mer captopurine) (6-MP), but is not limited thereto.

In addition, the antibiotic-based drugs are penicillin antibiotics, cephalosporins antibiotics, aminoglycosides antibiotics, macrolide antibiotics, sulfanamide antibiotics It may be selected from the group consisting of quinolone antibiotics and furane antibiotics, but is not limited thereto.

The natural-derived drugs include conventional herbal medicines and natural herbal medicines and drugs containing active ingredients obtained by modern extraction and separation methods, and the hormonal drugs include glucocorticoids, adrenal corticosteroids, and norepinephrine. (Norepinephrine), progesterone (progestogen), estrogen (estrogen) and androgen (androgen) may be selected from the group, but is not limited thereto.

According to another embodiment of the present invention, a decrease in leukocyte count, platelet count and neutrophil granulocyte count was confirmed in the animal model intraperitoneally administered or irradiated with cyclophosphamide (CYP), and the maghemite- As a result of oral administration of saponin nanoconjugates for 4 weeks, it was confirmed that the leukocyte count, platelet count and neutrophil granulocyte count, which are related to bone marrow suppression, in each animal model as shown in FIGS. 2 and 3. In particular, maghemite-saponin nanoconjugates showed better effects than the treatment with maghemite or saponin alone.

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

< Example  1> Maghemite -Saponin Nanoconjugate Synthesis

One. Maghemite Ginsenoside Rg3  Nanoconjugates Maghemite - ginsenoside Rg3 ) synthesis

To prepare maghemite combined with ginsenoside Rg3, 250 mg of ginsenoside Rg3 was dissolved in 50 mL of 30% ethanol. 2 g of maghemite was suspended in 200 mL of distilled water, washed twice, sonicated for 10 minutes using an UP100H ultrasonic apparatus (Hielscher, Germany), mixed with a ginsenoside Rg3 solution, and further sonicated for 10 minutes. .

Thereafter, the mixture of the mixture was vigorously mixed for at least 4 hours at room temperature, and then washed three times with distilled water in the maghemite-ginsenoside Rg3, followed by collecting the maghemite-ginsenoside Rg3 nanoconjugate using an external magnet. After lyophilization and stored at 4 ℃ until use.

2. Maghemite Ginsenoside F2 nanoconjugate ( Maghemite - ginsenoside  F2) Synthesis

To synthesize maghemite combined with ginsenoside F2, 250 mg of ginsenoside F2 was dissolved in 50 mL of 30% ethanol. 2 g of maghemite was suspended in 200 mL of distilled water, washed twice, sonicated for 10 minutes using an UP100H ultrasonic apparatus (Hielscher, Germany), mixed with a ginsenoside F2 solution, and further sonicated for 10 minutes. .

Thereafter, the maghemite-ginsenoside F2 obtained by vigorously mixing at room temperature for at least 4 hours was washed three times with distilled water, and the maghemite-ginsenoside F2 nanoconjugates were collected using an external magnet, and then lyophilized. It was stored at 4 ℃ until use.

3. Maghemite Ginsenoside CK nanoconjugate ( Maghemite - ginsenoside  CK) synthetic

To synthesize maghemite coupled with ginsenoside CK, 250 mg of ginsenoside CK was dissolved in 50 mL of 30% ethanol. 2 g of maghemite was suspended in 200 mL of distilled water, washed twice, sonicated for 10 minutes using an UP100H ultrasonic apparatus (Hielscher, Germany), mixed with a ginsenoside CK solution, and further sonicated for 10 minutes. .

Thereafter, the maghemite-ginsenoside CK obtained by vigorous mixing at room temperature for at least 4 hours was washed three times with distilled water, and the maghemite-ginsenoside CK nanoconjugates were collected using an external magnet and then lyophilized. It was stored at 4 ℃ until use.

4. Maghemite - Zipenoside XVII  Nanoconjugates Maghemite - gypenoside XVII ) synthesis

Gynostemma pentaphyllum ) 250 mg of fenenoside XVII was dissolved in 50 mL of 30% ethanol to synthesize maghemite combined with Gypenoside XVII, one of the dmarylene-based saponins extracted from pentaphyllum). 2 g of maghemite was suspended in 200 mL of distilled water, washed twice, sonicated for 10 minutes using an UP100H ultrasonic apparatus (Hielscher, Germany), and then mixed with phenenoside XVII solution and maghemite, followed by an additional 10 Ultrasonicated for a minute.

Thereafter, the maghemite-zipenoside XVII obtained by vigorous mixing for at least 4 hours at room temperature was washed three times with distilled water, and the maghemite-zipenoside XVII nanoconjugates were collected using an external magnet and then lyophilized. It was stored at 4 ℃ until use.

5. Maghemite -Ginseng Investigation  Nanoconjugates maghemite -Crude ginsenoside ) synthesis

In order to synthesize maghemite combined with ginseng checkonin, 250 mg of ginseng checkonin was dissolved in 50 mL of 30% ethanol. 2 g of maghemite was suspended in 200 mL of distilled water, washed twice, and sonicated for 10 minutes using an UP100H ultrasonic apparatus (Hielscher, Germany). Ginseng was mixed with the irradiated saponin solution and sonicated for an additional 10 minutes.

Thereafter, the maghemite-ginseng irradiated phosphonine obtained by vigorously mixing at room temperature for at least 4 hours was washed three times with distilled water, and the maghemite-ginseng irradiated phononin nanoconjugates were collected using an external magnet and then lyophilized. Stored at 4 ° C. until.

< Example  2> for taking Formulation example

Maghemite-saponin nanoconjugates stored as lyophilized powders as in Example 1 were prepared by dispersing to a dose using 5% carboxymethyl cellulose in the morning of the administration day. Dispersion was maintained using a mixer just prior to dosing to prevent precipitation after preparation.

< Example  3> Confirmation of erythrocyte improvement effect in anemia model

Five-week-old SD rats (Sprague-Dawley Rats) were purchased from central laboratory animals and used. Solid feed (Samyang blended feed for experimental animals, Samyang maintenance feed, Korea) and water for 1 week in the environment where the temperature is maintained at 22 ± 2 ℃ and humidity is 55 ~ 60% and the contrast cycle is controlled in 12 hours. It was freely adapted to the laboratory.

Seven rats each of the normal group, control group, experimental group 1 (maghemite), experimental group 2 (maghemite-ginsenoside Rg3), experimental group 3 (maghemite-ginsenoside F2), experimental group 4 (mag Hemite-Ginsenoside CK), Experimental Group 5 (Maghemite-Gifenoside XVII), Experimental Group 6 (Maghemite-Ginseng Irradiation Ponine), Experimental Group 7 (Ginsenoside Rg3), Experimental Group 8 (Ginsenoside F2 ), Experimental group 9 (ginsenoside CK), Experimental group 10 (Gifenoside XVII), Experimental group 11 (Ginseng probeponin), and comparative group (ferritin).

Iron deficiency anemia was induced by feeding the iron deficiency diet (TD.80396, Iron Deficient Diet, ENVIGO; g / kg diet) as shown in Table 1 to the experimental animals except the normal group.

Iron Deficient Diet (g / kg) Normal Diet (g / kg) Casein, low Cu & Fe 200.0 Casein, low Cu & Fe 200.0 DL-Methionine 3.0 DL-Methionine 3.0 Sucrose 549.99 Sucrose 549.99 Corn starch 150.0 Corn starch 150.0 Corn oil 50.0 Corn oil 50.0 Mineral Mix, Fe Deficient (81062) 35.0 Mineral mix 35.0 Vitamin mix 10.0 Vitamin mix 10.0 Choline bitartrate 2.0 Choline bitartrate 2.0 Ethoxyquin, antioxidant 0.01 Ethoxyquin, antioxidant 0.01

In order to confirm the induction of iron deficiency anemia according to the iron deficiency diet for 4 weeks, blood samples were collected from the jugular vein of each group of animals to confirm anemia-related blood markers. After confirming that anemia was induced, while feeding iron deficiency diet, Experimental group 1 was maghemite 12 mg / kg BW, Experimental group 2 was maghemite-ginsenoside Rg3 12 mg / kg BW, and experimental group 3 was maghemite- Ginsenoside F2 12 mg / kg BW, Experimental Group 4 is Maghemite-Ginsenoside CK 12mg / kg BW, Experimental Group 5 is Maghemite-Gifenoside XVII 12mg / kg BW, Experimental Group 6 is Maghemite-Ginseng 12 mg / kg BW of irradiated ponins, Ginsenoside Rg3 0.5mg / kg BW for Experimental Group 7, Ginsenoside F2 0.5mg / kg BW for Experimental Group 8, Ginsenoside CK 0.5mg / kg BW, Experimental Group 10 Norside XVII 0.5mg / kg BW, experimental group 11 was ginseng irradiated 0.5mg / kg BW, and the control group was orally administered 1 mL / kg BW of ferritin (ferroa syrup, Boryeong Pharmaceutical) for 4 weeks. Silver saline was orally administered in the same manner.

As shown in Table 2, the dose is based on the daily dose of the polysaccharide iron complex (150 mg as Fe), considering maghemite as the synthetic iron, according to the Conversion of Animal Doses to Human Equivalent Doses Based on Bodt Surface Area guidelines. Determined in terms of dose. In addition, the comparative group ferritin daily dose (10mL) was also determined in terms of rat dose in accordance with the guidelines.

Ginsenosides Rg3, F2, CK, Zipenoside XVII, ginseng probeponin was determined based on the analysis results of about 4 to 5% combined with maghemite.

Military classification Substance Dosage Normal 5% CMC - Control 5% CMC - Experimental group 1 Maghemite 12mg / kg Experiment group 2 Maghemite-Ginsenoside Rg3 12mg / kg Experiment group 3 Maghemite-Ginsenoside F2 12mg / kg Experimental Group 4 Maghemite-Ginsenoside CK 12mg / kg Experimental group 5 Maghemite-Gifenoside XVII 12mg / kg Experimental Group 6 Maghemite-Ginseng Research Ponine 12mg / kg Experimental group 7 Ginsenoside Rg3 0.5mg.kg Experimental Group 8 Ginsenoside F2 0.5mg / kg Experimental Group 9 Ginsenoside CK 0.5mg / kg Experimental group 10 Zipenoside XVII 0.5mg / kg Experimental Group 11 Ginseng 0.5mg / kg Comparison Ferricin 1mL / kg

As a result, the erythrocyte count and hemoglobin levels, which are the indicators of hemoglobin anemia, of the control group fed the iron deficiency diet were significantly reduced to 60 to 70% compared to the normal group fed the normal diet. As a result of oral administration of the sample for 4 weeks while maintaining the results, it was confirmed that the anemia index was restored to the normal level in the test groups ingested the maghemite-saponin nanoconjugates as shown in FIG. 1.

Specifically, in comparison with the control group, erythrocyte counts were 39% for maghemite-ginsenoside Rg3 nanoconjugates, 33% for maghemite-ginsenoside F2 nanoconjugates, 30% for maghemite-ginsenoside CK, Maghemite-Gifenoside XVII showed 26% improvement, and Maghemite-Ginseng researchponin improved 26%, whereas for non-nanocombinates, 23% for Maghemite and Ginsenoside Rg3 14%, Ginsenoside F2 was 10%, Ginsenoside CK was 5%, Zipenoside XVII was 6%, Ginseng probeponin was 16%, which was lower than that of the nanoconjugate. In the case of Chin, 31% of the effects were confirmed.

In addition, as a result of confirming the hemoglobin level compared to the control group, 47% of the maghemite-ginsenoside Rg3 nanoconjugates, 45% of the maghemite-ginsenoside F2 nanoconjugates, 38% of the maghemite-ginsenoside CK, Maghemite-Gifenoside XVII showed 31% improvement and Maghemite-Ginseng irradiated 30%, whereas 16% Maghemite and 24 Ginsenoside Rg3 were found for non-nanoconjugates alone. %, Ginsenoside F2 was 21%, ginsenoside CK 13%, giphenoside XVII 11%, ginseng probeponin 19% was found to have a lower effect on the nanoconjugates, ferritin treated with the control group Showed a 32% effect.

From the above results, the effect of improving the erythrocyte and hemoglobin levels of the maghemite-saponin nanoconjugates was similar to that of the normal control group, and it was confirmed that the improvement of each of the substances alone was achieved.

< Example  4> Identify the effects of chemotherapy-induced bone marrow suppression

Five-week-old SD rats (Sprague-Dawley Rats) were purchased from central laboratory animals and used. Solid feed (Samyang blended feed for experimental animals, Samyang maintenance feed, Korea) and water for 1 week in the environment where the temperature is maintained at 22 ± 2 ℃ and humidity is 55 ~ 60% and the contrast cycle is controlled in 12 hours. The breeding was freely supplied.

To confirm the hematopoietic inhibitory activity of maghemite-saponin nanoconjugates in animal models, hematopoietic disorders were induced in experimental animals as follows.

Cyclophosphamide (hereinafter referred to as "CYP") (purchased from Sigma) was used as a hematopoietic inducer. CYP is an alkylating agent used as an anticancer chemotherapeutic agent and an immunosuppressive agent, and bone marrow suppression has been reported as a common side effect. It is used in the form of powder for injection and tablets for oral administration, and is widely used for malignant tumors, non-Hodgkin's lymphoma, and breast cancer. Because of myelosuppression side effects, myelosuppression is contraindicated. However, CYP is classified as a drug having high inhibitory effect on platelets and relatively low side effects on platelet function, and is also used as an effect of treating platelet reduction in immune thrombocytopenia disease (Ghanima W et. , al, blood.2012, Reiner A et., al, blood. 1995). Administration of cytotoxic agents, such as CYP, promotes the production of stem cells into peripheral blood.

The experimental animals were divided into 14 groups (7 for each group) as shown in Table 3, and CP was administered 150 mg / kg BW to each group except the normal group on the first day of the experiment to induce hematopoietic dysfunction.

Military classification Substance Dosage Normal 5% CMC - Control 5% CMC - Experimental group 1 Maghemite 12mg / kg Experiment group 2 Maghemite-Ginsenoside Rg3 12mg / kg Experiment group 3 Maghemite-Ginsenoside F2 12mg / kg Experimental Group 4 Maghemite-Ginsenoside CK 12mg / kg Experimental group 5 Maghemite-Gifenoside XVII 12mg / kg Experimental Group 6 Maghemite-Ginseng Research Ponine 12mg / kg Experimental group 7 Ginsenoside Rg3 0.5mg.kg Experimental Group 8 Ginsenoside F2 0.5mg / kg Experimental Group 9 Ginsenoside CK 0.5mg / kg Experimental group 10 Zipenoside XVII 0.5mg / kg Experimental Group 11 Ginseng 0.5mg / kg Comparison Ferricin 1mL / kg

Thereafter, daily experimental group 1 was 12 mg / kg BW of maghemite, experimental group 2 was 12 mg / kg BW of maghemite-ginsenoside Rg3, experimental group 3 was 12 mg / kg BW of maghemite-ginsenoside F2, experimental group 4 is Maghemite-Ginsenoside CK 12mg / kg BW, Experimental Group 5 is Maghemite-Gifenoside XVII 12mg / kg BW, Experimental Group 6 is Maghemite-Ginseng probeponin 12mg / kg BW, Experiment 7 is Gene Cenoside Rg3 0.5mg / kg BW, Experimental Group 8 is Ginsenoside F2 0.5mg / kg BW, Experimental Group 9 is Ginsenoside CK 0.5mg / kg BW, Experimental Group 10 is Zipenoside XVII 0.5mg / kg BW, Experimental Group 11 Silver ginseng probeponin 0.5mg / kg BW, the control group was orally administered 1mL / kg BW of ferritin (ferroa syrup, Boryeong Pharmaceuticals) for 4 weeks. The dosage was determined based on the daily dose and content of each substance as in the iron deficiency anemia model.

As a result, as shown in FIG. 2, the hematopoietic function index, leukocyte count, platelet count, and neutrophil granulocyte count of the control group treated with cyclophosphamide (CYP) were significantly decreased as compared with the normal group ingesting the normal feed as described above. As a result of treating the maghemite-saponin nanoconjugates, a significant improvement was shown by the oral administration of each maghemite-saponin nanoconjugate.

More specifically, the leukocyte count compared to the control group was 59% for maghemite-ginsenoside Rg3 nanoconjugates, 49% for maghemite-ginsenoside F2 nanoconjugates, 46% for maghemite-ginsenoside CK, and Maghe Mite-zipnoside XVII was 43% and maghemite-ginseng irradiated with 20% improvement, whereas maghemite 20% and ginsenoside Rg3 were 35% when the non-nano binder was administered alone. %, Ginsenoside F2 was 22%, Ginsenoside CK was 16%, Gifenoside XVII was 17%, Ginseng probeponin was 18%, showing a lower effect than the nanoconjugate. In the case of Chin, 20% of the effects were found.

From the above results, it was confirmed that the nanoconjugates were superior to the case where the substance was administered alone, and exhibited an effect of improving leukocyte count equal to or higher than that of the comparative group.

In addition, the leukocyte count compared to the control group was 59% for maghemite-ginsenoside Rg3 nanoconjugates, 49% for maghemite-ginsenoside F2 nanoconjugates, 46% for maghemite-ginsenoside CK, and maghemite -43% for Zipenoside XVII and 20% for Maghemite-Ginseng irradiated, whereas 20% for Maghemite, 35% for Ginsenoside Rg3 for non-nanoconjugates alone. Ginsenoside F2 is 22%, Ginsenoside CK is 16%, Zipenoside XVII is 17%, Ginseng irradiated with 18%, which is lower than the nanoconjugates. The effect was 20%.

From the above results, it was confirmed that the nanoconjugates were superior to the individual substances and showed an effect of improving leukocyte levels equal to or greater than that of the comparative group.

Platelet counts compared to the control group were 55% for maghemite-ginsenoside Rg3 nanoconjugates, 51% for maghemite-ginsenoside F2 nanoconjugates, 45% for maghemite-ginsenoside CK, and maghemite-zippe. Norside XVII showed 36% improvement and maghemite-ginseng irradiated 40%, whereas for non-nanocombinates, 21% for maghemite alone, 25% for ginsenoside Rg3, and ginsenosides. Side F2 was 23%, Ginsenoside CK was 24%, Zipenoside XVII was 22%, and ginseng irradiated saponin was 25%, which was lower than that of nanoconjugates. It showed an improvement effect.

The results were also confirmed that the nano-conjugates showed better platelet counts than the individual substances as well as the white blood cell counts identified above.

In addition, the neutrophil granulocyte count compared to the control group was 49% for maghemite-ginsenoside Rg3 nanoconjugates, 43% for maghemite-ginsenoside F2 nanoconjugates, 38% for maghemite-ginsenoside CK, and mag Hematite-Gifenoside XVII increased by 33% and Maghemite-Ginseng searchponin increased by 26%, whereas Maghemite 32%, Ginsenoside Rg3 18%, Gene Cenoside F2 was 14%, Ginsenoside CK was 15%, Zipenoside XVII was 15%, and ginseng probeonin was 18%, showing a lower growth rate than maghemite-saponin nanoconjugates. Chin showed an increase of 26%.

From the above results, it was confirmed that maghemite-saponin nanoconjugates can alleviate or prevent myelosuppressive side effects caused by cyclophosphamide, an anticancer drug. It may be used simultaneously and may have the effect of promoting bone marrow hematopoiesis to restore the white blood cell, platelet and neutrophil granulocyte count to the normal range.

< Example  5> Identify the effects of bone marrow suppression induced by radiation therapy

In order to confirm the effect of improving the bone marrow suppression induced by radiation therapy, each sample was orally administered to mice irradiated with Co at a dose of 5 Gy as shown in Table 4.

Military classification Substance Dosage Normal 5% CMC - Control 5% CMC - Experimental group 1 Maghemite 12mg / kg Experiment group 2 Maghemite-Ginsenoside Rg3 12mg / kg Experiment group 3 Maghemite-Ginsenoside F2 12mg / kg Experimental Group 4 Maghemite-Ginsenoside CK 12mg / kg Experimental group 5 Maghemite-Gifenoside XVII 12mg / kg Experimental Group 6 Maghemite-Ginseng Research Ponine 12mg / kg Experimental group 7 Ginsenoside Rg3 0.5mg.kg Experimental Group 8 Ginsenoside F2 0.5mg / kg Experimental Group 9 Ginsenoside CK 0.5mg / kg Experimental group 10 Zipenoside XVII 0.5mg / kg Experimental Group 11 Ginseng 0.5mg / kg Comparison Ferricin 1mL / kg

Thereafter, experimental group 1 was 12 mg / kg BW of maghemite, experimental group 2 was 12 mg / kg BW of maghemite-ginsenoside Rg3, experimental group 3 was 12 mg / kg BW of maghemite-ginsenoside F2, experimental group 4 Is Maghemite-ginsenoside CK 12mg / kg BW, Experimental Group 5 is Maghemite-Gifenoside XVII 12mg / kg BW, Experimental Group 6 is Maghemite-Ginseng Irradiation 12mg / kg BW, Experimental Group 7 is Ginseno Side Rg3 0.5mg / kg BW, experimental group 8 is ginsenoside F2 0.5mg / kg BW, experimental group 9 is ginsenoside CK 0.5mg / kg BW, experimental group 10 is fenenoside XVII 0.5mg / kg BW, experimental group 11 is Ginseng irradiated with 0.5mg / kg BW, ferrichin (ferroa syrup, Boryung Pharmaceutical) 1mL / kg BW was administered orally for 5 weeks each week for 4 weeks. The dosage was determined based on the daily dose and content of each substance as in the iron deficiency anemia model.

As a result, as shown in FIG. 3, the hematopoietic function, platelet count, and neutrophil granulocyte count, which are indicators of hematopoietic function, of the control group irradiated with radiation were significantly decreased, and in the experimental group to which the maghemite-saponin nanoconjugate was orally administered, Significant improvement was confirmed.

Specifically, the leukocyte count compared to the control group was 56% for the maghemite-ginsenoside Rg3 nanoconjugates, 47% for the maghemite-ginsenoside F2 nanoconjugates, 43% for the maghemite-ginsenoside CK, and 43% for the maghemite -41% for Zipenoside XVII and 43% for Maghemite-Ginseng probeponin. On the other hand, for non-nanocombinates, 18% of maghemite alone, 32% of ginsenoside Rg3, 20% of ginsenoside F2, 14% of ginsenoside CK, 15% of gifenoside XVII, ginseng Irradiation was 15% lower than that of the nanoconjugates. The ferritin treated with the control group also showed an increase of 18%, indicating that the nanoconjugates were superior to the individual substances and showed more than equivalent white blood cell improvement.

Platelet levels compared to the control group were 53% for maghemite-ginsenoside Rg3 nanoconjugates, 49% for maghemite-ginsenoside F2 nanoconjugates, 43% for maghemite-ginsenoside CK, and maghemite-zippe. Norside XVII showed 34% improvement, and maghemite-ginseng irradiated with 38% improvement, compared to 19% for maghemite, 23% for ginsenoside Rg3, and ginsenoside for non-nanoconjugates alone. F2 was 22%, Ginsenoside CK was 22%, Zipenoside XVII was 21%, Ginseng irradiated saponin was 24%, which was lower than that of nanoconjugates. The effect was shown.

From the above results, it was confirmed that the maghemite-saponin nanoconjugates showed better platelet count-improving effects than the individual substances as in the white blood cell count.

On the other hand, the neutrophil granulocyte values compared to the control group was 54% for the maghemite-ginsenoside Rg3 nanoconjugates, 47% for the maghemite-ginsenoside F2 nanoconjugates, 42% for the maghemite-ginsenoside CK, and mag Hematite-Gifenoside XVII showed 38% improvement, and Maghemite-Ginseng irradiated 30%, while 22% Maghemite and Ginsenoside Rg3 were obtained when treated with individual substances other than nanoconjugates. 22%, Ginsenoside F2 was 18%, Ginsenoside CK was 19%, Gifenoside XVII was 18%, and Ginseng probeponin was 22%, which was lower than that of the nanoconjugate. Chin showed a 36% effect.

From the above results, the maghemite-saponin nanoconjugates can alleviate the bone marrow suppression side effects induced by anti-cancer therapy, which is an anticancer treatment method. Or may be used for prevention, and may promote the bone marrow hematopoiesis to restore the white blood cell, platelet and neutrophil granulocyte count to the normal range.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific description is merely a preferred embodiment, thereby not limiting the scope of the present invention. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (13)

A pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient. The pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders according to claim 1, wherein the nanoconjugate is directly bound to saponin on the surface of maghemite. The pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders according to claim 1, wherein the saponin is a dmarylene-based saponin isolated from a plant. The pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders according to claim 3, wherein the dammarene-based saponin is selected from the group consisting of ginseng-derived ginsenosides and dodol-derived fenenosides. The pharmaceutical composition for preventing or treating hematopoietic dysfunction diseases according to claim 4, wherein the ginseng-derived ginsenoside is selected from the group consisting of ginsenoside Rg3, ginsenoside F2, and ginsenoside CK. The pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders according to claim 4, wherein the dodol-derived fenenoside is zipenoside XVII. The pharmaceutical composition for preventing or treating hematopoietic dysfunction disorders according to claim 1, wherein the maghemite-saponin nanoconjugate increases the number of red blood cells, white blood cells, platelets or neutrophil granulocytes. The method according to claim 1, wherein the hematopoietic dysfunction disorder in the group consisting of anemia, malignant lymphoma, leukemia, myeloid thrombocytopenia, idiopathic thrombocytopenic purpura, thrombocytopenia, lymphocytosis, mononucleosis, granulocytopenia and myelodysplastic syndrome Pharmaceutical composition for preventing or treating hematopoietic dysfunction disorder, characterized in that selected. Health food for preventing or improving hematopoietic dysfunction disorders containing maghemite-saponin nanoconjugates as an active ingredient. It contains maghemite-saponin nanoconjugate as an active ingredient, and the nanoconjugate is an adjuvant for preventing or improving hematopoietic dysfunction which prevents or improves the reduction of the number of erythrocytes, leukocytes, platelets or neutrophil granulocytes by chemical or radiation therapy. . The adjuvant of claim 10, wherein the nanoconjugate is directly bound to saponin on the surface of maghemite. The adjuvant of claim 10, wherein the chemical is selected from the group consisting of alkylating agents, anti-metabolic agents, antibiotics, drugs derived from natural products, and hormones. The adjuvant according to claim 10, wherein the adjuvant is administered in combination with a chemical or radiation.

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