KR100530369B1 - The injection system of nanoparticles bound antitumor agents - Google Patents

Abstract

The present invention relates to a drug system of an injection formulation in which nanoparticles are coupled to an anticancer substance, and more particularly, by synthesizing an effective drug combining silica nanoparticles to an existing anticancer substance and injecting the same into a cancer cell by silica nanoparticles. The present invention relates to a drug system and an anticancer composition capable of maximizing anticancer activity since the anticancer substance is supported to prevent the drug from being diffused into normal cells.

Description

The injection system of nanoparticles bound antitumor agents

The present invention relates to a drug system of an injection formulation in which nanoparticles are coupled to an anticancer substance, and more particularly, by synthesizing an effective drug combining silica nanoparticles to an existing anticancer substance and injecting the same into a cancer cell by silica nanoparticles. The present invention relates to a drug system and an anticancer composition capable of maximizing anticancer activity since the anticancer substance is supported to prevent the drug from being diffused into normal cells.

Existing anticancer drugs can spread to other tumor cells immediately even when administered to tumor cell sites, causing various side effects. Therefore, recently, anti-cancer drug delivery systems, etc. have been developed to solve this side effect, and use of a mixture of polymers prevents the spread of drugs to areas other than the cancer site, thereby reducing various side effects of conventional anti-cancer drugs. No. 2002-23441], ionic strength, pH sensitivity and instability problems.

In addition, a method for delivering an anticancer agent using liposomes is already known [Domestic Patent Publication No. 2001-24232], but there is a problem that anticancer activity is not exerted to the maximum by attacking not only cancer cells but also normal cells.

Thus, the present inventors have studied to solve the above problems, as a result of injection into cancer cells to treat the effective drug to combine the silica nanoparticles with the existing anticancer substance by the anticancer substance is fixed by the silica nanoparticles normal cells The present invention has been completed by developing a drug system to prevent the drug from spreading to maximize the anticancer activity.

Accordingly, an object of the present invention is to provide a drug system of a new injection formulation having excellent anticancer effect and a method for preparing the same.

The present invention, as shown in the following Structural Formula 1, the cell attachment portion and the anti-cancer substance portion and the cell to exhibit the anti-cancer activity to maximize the anti-cancer substance portion and the anti-cancer substance is fixed to the cancer cells having anticancer activity It is characterized by a drug system of the injection formulation consisting of a crosslinking portion connecting the attachment portion and a method for preparing the same.

[Formula 1]

In addition, the drug system consisting of the compound represented by the following structural formula 4 except for the anti-cancer substance portion and a method for preparing the same.

[Structure 4]

Referring to the present invention in more detail as follows.

Drug system according to the present invention can be divided into three parts, the structure is shown in the following structural formula (1).

Structural Formula 1

As shown in Structural Formula 1, it is divided into a cell attachment part, a crosslinking part, and an anticancer material part consisting of nanoparticles. It acts as a crosslinking link. It depends on the functional groups in the anticancer substance, (X represents O, NH, S, or COO). In this case, the cell attachment part is selected from silica, alumina, and TiO ₂ , preferably 5 to 900 nm, because when the size of the nanoparticles is less than 5 nm, there is a risk of movement of the nanoparticles in the cell and exceeds 900 nm. This is because it is too sparsely present in the cell.

The preparation method of the drug system is described in detail as follows.

First, a compound represented by the following structural formula 2 is synthesized by reacting an anticancer substance having an active group selected from a hydroxyl group, an amino group, a thiol group, and a carboxy group with 3- (triethoxysilyl) propyl isocyanate. At this time, as an anticancer substance, Uracil, 5-Fluorouracil, Tegafur, Vinoelbine bitartrate, Methotrexate, Carboplatine ), Cisplatin, Oxaliplatine, Cytosine arabinoside HCl, Mitoxantrone HCl, Tamoxifen Citrate, Nimustine HCl, Nimustine HCl), DaunorubicinHCl (DaunorubicinHCl), EpirubicinHCl (EpirubicinHCl), IdarubicinHCl, IdarubicinHCl, DoxorubicinHCl (DoxorubicineHCl), Doxyfluidine (Doxifludine), Dacarbazine, Thioguanine, Formestane, Leuprorelin acetate, Leuprolide. Chlorambucil, Busufan, Megesterol acetate, Tretinoin, Vinblastine sulfate, Vincristine sulfate, Tenniposide, Eto Ectoside HCl Carmustine (BCNU), Lomustine (CCNU), Estramustine, Ranimustine, Cytarabine, Cyclophos Pyamide-HCl (Cyclophosphamide / HCl), Bicaltamide, Iphosamide (Ifosfamide), Flutamide, Melphalan, Docetaxel, Pacitaxel, Paclitaxel, Doc Dactimomycin, Mercaptopurine, 6-Mercaptopurine, Aldesleukin, Hydroxyurea, Topotecan HCl, Altre Altretamine, Medroxyprogesterone Acetate (Medrox) yprogesterone acetate, Polysaccharide K, Polysaccharide Phellinus Linteus, Polypeptide, Tegafur + Uracil Mustard, BCG strain Tice, Interferon γ, DNA recombinant Interferon alpha, L-Asparaginase, Betulinic acid, Camptothecin, Colchiceine ), Irinotecan, Lapachol, Ladochol, Podophyllotoxin, Taxol, Teneniposide, Vinblastine, Vincristine and Etoposide Etc. are preferable.

[Formula 2]

In Formula 2, X represents O, NH, S, or COO.

The drug system is prepared by synthesizing the compound represented by the following Structural Formula 3 by adding water to the compound synthesized as described above (compound represented by Structural Formula 2) to form nanoparticles for cell adhesion (see Scheme 1 and Scheme 2).

In addition, after synthesizing the compound represented by the formula 2 and the compound and tetraethoxysilane in a 1: 1 molar ratio can be synthesized the compound represented by the following formula 3, wherein the compound represented by the formula 3 directly injected into cancer cells The anti-cancer substance is fixed to cancer cells by nanoparticles.

Structural Formula 3

In Structural Formula 3, X represents O, NH, S, or COO.

In the second reaction step of Scheme 1, the triethoxysilin material in which the anticancer substance is bound reacts with water to form nanoparticles in which the anticancer substance is bound, wherein the amount of water is controlled between 1 and 30 moles of nanoparticles. The size can be adjusted from 5 to 900 nm.

In addition, tetraethoxysilane, acetonitrile, dimethyl sulfoxide, and the like are used as a solvent used to form the cell adhesion nanoparticles in the second reaction step of Scheme 2, and water is used as a reaction material. By controlling the ratio of the compound represented by 2 and the tetraethoxysilane and the amount of water, respectively, between 1 and 30 moles, nanoparticles having a size of 5 to 900 nm can be produced. NH ₄ OH or an acid may be used as the reaction catalyst, and as the acid used, a weak acid such as CH ₃ COOH is preferable. However, even without this catalyst, silica nanoparticles are slowly formed.

Meanwhile, in the drug system of Formula 1, except for the anticancer substance, the compound of Formula 4 is reacted with 3- (triethoxysilyl) propyl isocyanate directly with water to form a -CH ₂ CH ₂ CH ₂ NHCOOH group as shown in Scheme 3 below. Silica nanoparticles can be produced and the size of the nanoparticles can be adjusted to 5 to 900 nm depending on the number of moles of water.

[Structure 4]

The drug system thus prepared is a new drug system that maximizes the anticancer effect by combining nanoparticles with existing anticancer substances and injecting them directly into cancer cells, thereby preventing the diffusion of drugs into normal cells by fixing the anticancer substances by silica nanoparticles. As a result, it was confirmed that it is very useful for suppressing general cancer cell growth. In addition, it was confirmed that the drug system except for the anticancer substance also shows anticancer effects.

The effective dose of the active ingredient may vary depending on the age, physical condition, weight, etc. of the patient, but is generally administered within the range of 1 to 100 mg / kg (weight) / day. In addition, the dose is administered once a day or divided several times a day within the effective dosage range per day.

delete

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

Example 1 Synthesis of Silica Nanoparticles (in Acid Solution)

Tetraethoxysilane (0.1 mole) was dissolved in anhydrous ethanol (40 mL), 0.1 N HCl (5.0 mL) was added thereto, and then heated to 38 ° C. for 24 hours, followed by cooling to room temperature. 10 ml of concentrated hydrochloric acid (12N) was added again, followed by stirring for 24 hours. After cooling the solution, the nanoparticles were collected by centrifugation (10000 rpm) [yield 90%>, BET = 1234 m ² / g, SEM = 7-20 nm].

Example 2 Synthesis of Silica Nanoparticles (in Neutral Solution)

Tetraethoxysilane (0.1 mole) was dissolved in anhydrous ethanol (40 mL), distilled water (5.0 mL) was added thereto, and then heated to 38 ° C. for 24 hours, followed by cooling to room temperature. After cooling the solution, the nanoparticles were collected by centrifugation (10000 rpm) [yield 90%>, BET = 1234 m ² / g, SEM = 7-20 nm].

Example 3: Synthesis of Silica Nanoparticles (in Basic Solution)

Ammonium hydroxide (15 mL, 29%), distilled shoe (129 mL), anhydrous ethanol (330 mL) were dissolved in each other, and 3- (triethoxysilyl) propyl isocyanate (0.1 mole) was immediately added to the solution. Stir at room temperature for 24 hours. Then, tetraethoxysilane (0.1 mole) was added to the above solution and then stirred at room temperature to form particles of 7-20 nm [yield 90%>, SEM = 7-20 nm].

Example 4 Synthesis of Silica Nanoparticles Containing Anticancer Agent (Hydroxyurea)

After dissolving 3- (triethoxysilyl) propyl isocyanate (0.025 mole) in anhydrous THF (20 mL) and stirring at room temperature, hydroxyurea (0.025 mole) was dissolved in anhydrous THF and added dropwise to the above reaction material. While stirring at room temperature. Stirring at about 38 ℃ for 24 hours to obtain the material of formula 2 (yield 90%>). Then distilled water (23 ㎖), THF (25 ㎖) the added, then the mixture was stirred at room temperature to synthesize the material of the Structure 3 [yield: 90%>, SEM = 25 nm , IR (KBr): OH (3400 cm - ¹ , s), sat. CH (2900 cm ⁻¹ , m), C═O (1700 cm ⁻¹ , s)].

Example 5 Synthesis of Silica Nanoparticles Conjugated with Anticancer Agent (6-Mercaptopurine)

After dissolving 3- (triethoxysilyl) propyl isocyanate (0.04 mole) in acetone (35 ml) and stirring at room temperature, 6-mercaptopurine-H ₂ O (0.04 mole) was dissolved in acetone, and then Stir at room temperature with dropwise addition to the reaction. Stirring at about 100 ℃ for 24 hours to obtain the material of formula 2 (yield 90%>). Distilled water (18 mL) and acetone (20 mL) were then added, followed by stirring at room temperature to synthesize the material of Structural Formula 3 [yield 90%>, SEM = 50 nm, IR (KBr): NH (3400 cm ^{−). 1} , s), sat. CH (2900 cm ⁻¹ , s), C═O (1600 cm ⁻¹ , s)].

Example 6: Synthesis of Silica Nanoparticles Conjugated with Anticancer Agent (Cytocin Arabinoside-HCl)

After dissolving 3- (triethoxysilyl) propyl isocyanate (0.0036 mole) in DMSO (20 mL) and stirring at room temperature, cytosine arabinoside-HCl (0.0036 mole) was dissolved in DMSO, Stir at room temperature with dropwise addition. Stirring at about 100 ℃ for 24 hours to obtain the material of formula 2 (yield 90%>). Distilled water (3.2 mL) and DMSO (10 mL) were added, followed by addition of tetraethoxysilane (0.0036 mole), followed by stirring at room temperature to synthesize the material of Structural Formula 3 [yield 90%>, SEM = bulk, IR (KBr): NH (3400 cm ⁻¹ , m), sat. CH (2900 cm ⁻¹ , m)].

Example 7 Synthesis of Silica Nanoparticles Conjugated with Anticancer Agent (Cyclophosphonide-HCl)

After dissolving 3- (triethoxysilyl) propyl isocyanate (0.02 mole) in DMSO (20 ml) and stirring at room temperature, cyclophosphamide-H ₂ O (0.01 mole) was dissolved in DMSO, followed by reaction Stir at room temperature with dropwise addition to the material. Stirring at about 80 ℃ for 24 hours to obtain the material of formula 2 (yield 90%>). Distilled water (9 mL) and DMSO (10 mL) were then added, followed by stirring at room temperature to synthesize the material of Structural Formula 3 [yield 90%>, SEM = 25 nm, IR (KBr): NH (3400 cm ^{−). 1} , m), sat. CH (2900 cm ⁻¹ , m)].

Example 8 Synthesis of Silica Nanoparticles Containing Anticancer Agent (Uracil)

Dissolve 3- (triethoxysilyl) propyl isocyanate (0.05 mole) in DMSO (30 mL) and stir at room temperature, while dissolving uracil (0.05 mole) in DMSO and stir at room temperature dropwise to the reaction product It was. Stirring at about 80 ° C afforded the material of formula 2 (yield 90%>). Distilled water (18 mL) and DMSO (20 mL) were added, followed by stirring at room temperature to synthesize the material of Structural Formula 3 [yield 90%>, SEM = 25 nm, IR (KBr): NH (3100 cm ^-1). , s), sat.CH (2900 cm ⁻¹ , s), C═O (1700 cm ⁻¹ , s), C = N (1650 cm ⁻¹ , s)].

Example 9: -CH ₂ CH ₂ CH ₂ Synthesis of NHCOOH-bound Silica Nanoparticles

3- (triethoxysilyl) propyl isocyanate (0.05 mole) was dissolved in DMSO (30 mL) and stirred at room temperature. Distilled water (18 mL) and DMSO (20 mL) were added thereto, followed by stirring at room temperature to combine -CH ₂ CH ₂ CH ₂ NHCOOH to synthesize silica nanoparticles of Structural Formula 4 [yield 90%>, SEM = 50 nm, IR (KBr): sat.CH (2900 cm ⁻¹ , m), C = N (1650 cm ⁻¹ , m), C═O (1600 cm ⁻¹ , m)]. In Structural Formula 4, the silica nanoparticles are triethoxysilyl groups.

Example 10 Injection Preparation

       A: hydroxyurea (0.013 mole) was dissolved in DMSO (15 mL).

       B: 3- (triethoxysilyl) propyl isocyanate (0.013 mole) was dissolved in DMSO (15 mL).

C: DMSO (10 mL) / H ₂ O (10 mL)

       A and B mixed first, then mixed with C and injected into affected area

Example 11: Injection Preparation

A: Cyclophosphamide-HCl (0.0036 mole) was dissolved in acetonitrile (15 mL).

       B: 3- (triethoxysilyl) propyl isocyanate (0.0072 mole) was dissolved in DMSO (15 mL).

C: DMSO (1.0 mL) / H ₂ O (1.0 mL)

       A and B mixed first, then mixed with C and injected into affected area

Example 12 Injection Preparation

A: Uracil (0.0089 mole) was dissolved in DMSO (15 mL).

       B: 3- (triethoxysilyl) propyl isocyanate (0.0089 mole) was dissolved in DMSO (15 mL).

C: DMSO (4 mL) / H ₂ O (3 mL)

       A and B mixed first, then mixed with C and injected into affected area

Example 13: Toxicity Test

Toxicity test was performed as follows for the anticancer agent according to the present invention.

Each of the materials synthesized in Examples 4 to 8 was dissolved in distilled water, diluted with water, and then administered to the mice (10 mice per group) at 100 mg / kg, respectively, and observed for 7 days, but no rats died.

Test Example: Confirmation of Anticancer Effect

The new anticancer substances obtained in Examples 4 to 9 were injected around cancer cells to show the cell growth rate (%) in Tables 1 to 6 below.

A549: lung cancer cells

       SK-OV-3: Uterine Cancer Cells

       SK-MEL-2: skin cancer cells

       XF498: central nervous system cancer cells

       HCT15: Colon Cancer Cell

Concentration (µg / ml) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 4 0.1 103.1034 91.8485 105.0623 100.1347 100.8493 0.3 104.4828 87.7596 103.1184 99.1244 97.0276 One 95.4680 90.6166 101.8224 98.6632 96.3631 3 91.6256 86.0822 95.6667 98.5725 90.1507 10 68.6207 81.5477 95.5047 97.6164 85.9108 30 42.8079 70.8276 81.3709 95.4041 65.1805

Concentration (µg / ml) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 5 0.1 68.7533 97.0616 73.9446 72.8690 86.3636 0.3 65.3050 95.1223 43.3254 25.9188 73.5178 One 23.0769 70.4106 41.9147 20.5696 33.7945 3 19.5225 45.2288 40.5041 19.6459 9.2885 10 11.6711 22.9854 36.8945 19.0302 -7.5521 30 8.5411 6.5893 6.4827 10.7177 -39.3229

Concentration (µg / ml) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 6 0.1 90.9515 102.0575 103.0335 99.8627 98.1567 0.3 87.3134 98.0601 97.1182 97.7339 95.0845 One 84.6549 96.4141 98.1041 94.8240 88.9401 3 76.1194 91.4174 91.2030 89.9871 71.5822 10 34.3284 78.6612 82.1405 81.1073 49.1551 30 8.4422 62.1133 39.7099 57.2189 30.4147

Concentration (µg / ml) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 7 0.1 91.6135 99.0217 98.7222 98.2755 96.5443 0.3 86.1206 101.3340 99.9201 97.3524 88.5529 One 73.0750 97.7470 93.2515 95.9756 59.3952 3 65.6204 95.4643 93.9303 95.0009 69.1145 10 56.2040 80.0193 80.6329 93.8257 65.8747 30 46.6405 65.7304 54.2777 91.4152 55.9395

Concentration (µg / ml) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 8 0.1 104.7344 100.4001 99.6358 96.0474 98.5586 0.3 104.1557 99.5731 99.3121 95.2404 96.0360 One 87.8485 98.0288 95.5083 93.3751 92.6126 3 82.8511 93.7987 88.4219 92.7963 81.4414 10 82.3777 88.7976 86.6464 91.5068 76.3964 30 76.2230 79.6528 53.7481 89.0826 75.2973

Concentration 10% (w / v) A549 SK-OV-3 SK-MEL-2 XF498 HCT15 Example 9 × 10 ⁶ dilution 109.3776 102.7013 104.8950 93.8719 107.2979 × 10 ⁵ dilution 109.6567 103.8171 102.4475 93.4540 89.0532 × 10 ⁴ dilution 102.0653 100.6753 95.2373 88.3937 88.7417 × 10 ³ dilution 99.4418 89.2241 86.1750 82.9619 8435234 × 100 dilution 74.5465 60.0382 18.1082 64.0669 33.8157 × 10 dilution -43.4599 -84.0236 -47.9647 -74.5935 -72.8000

(In the above table, you can see that the value decreases as the concentration increases, and this value has a range of +100 to -100. If the value is +100, it indicates the value of the original sample. Also, a value of -100 indicates that all cells are dead (100%).)

As shown in Table 1 to Table 6, the drug system according to the present invention was confirmed that the anticancer effect was excellent.

As described above, the drug system according to the present invention binds nanoparticles serving as supporters to existing anticancer substances and directly injects them into cancer cells, whereby the anticancer substances are immobilized to the cancer cells by the nanoparticles, thereby spreading drugs to normal cells. It is developed to maximize the anti-cancer effect in the cancer cell area to be treated to prevent.

Claims (13)

As represented by the following structural formula 3, Silica nanoparticle part to maximize the anticancer activity, An anticancer substance moiety having an active group selected from a hydroxy group, an amino group, a thiol group, and a carboxyl group; Drug system of the injection formulation, characterized in that consisting of a cross-linking portion connecting the silica nanoparticle portion and the anticancer substance portion; [Formula 3] In formula 3, X represents O, NH, S or COO. The drug system according to claim 1, wherein the nanoparticles are 5 to 900 nm in size. delete According to claim 1, wherein the anticancer material is Uracil (5-rafluorouracil), 5-Fluorouracil (Tegafur), Vinoelbine bitartrate (Vinorelbine bitartrate), Methotrexate (Carthotrexate), Carr Carboplatine, Cisplatine, Oxaliplatine, Cytocine arabinoside HCl, Mitoxantrone HCl, Tamoxifen Citrate, Tamoxifen Citrate, Nimustine HCl (Nimustine HCl), Daunorubicin HCl (Daunorubicin HCl), Epirubicin HCl (Epirubicin HCl), Idarubicin HCl (Idarubicin HCl), Doxorubicin HCl (Doxorubicine HCl) Doxifludine, Dacarbazine, Thioguanine, Formestane, Leuprorelin acetate, Leuprolide. Chlorambucil, Busufan, Megesterol acetate, Tretinoin, Vinblastine sulfate, Vincristine sulfate, Tenniposide, Eto Ectoside HCl Carmustine (BCNU), Lomustine (CCNU), Estramustine, Ranimustine, Cytarabine, Cyclophos Pyamide-HCl (Cyclophosphamide / HCl), Bicaltamide, Iphosamide (Ifosfamide), Flutamide, Melphalan, Docetaxel, Pacitaxel, Paclitaxel, Doc Dactimomycin, Mercaptopurine, 6-Mercaptopurine, Aldesleukin, Hydroxyurea, Topotecan HCl, Altre Altretamine, Medroxyprogesterone Acetate (Medrox) yprogesterone acetate, Polysaccharide K, Polysaccharide Phellinus Linteus, Polypeptide, Tegafur + Uracil Mustard, BCG strain Tice, Interferon γ, DNA recombinant Interferon alpha, L-Asparaginase, Betulinic acid, Camptothecin, Colchiceine ), Irinotecan, Lapachol, Ladochol, Podophyllotoxin, Taxol, Teneniposide, Vinblastine, Vincristine and Etoposide Drug system, characterized in that selected from. delete 1) synthesizing a compound represented by the following Structural Formula 2 by reacting an anticancer substance having an active group selected from a hydroxyl group, an amino group, a thiol group, and a carboxy group with 3- (triethoxysilyl) propyl isocyanate; And 2) The method of preparing a drug system of the structural formula 3 of the injection formulation comprising the step of synthesizing the compound represented by the following structural formula 3 by adding water to the compound represented by the structural formula 2. [Formula 2] In the formula 2; X represents O, NH, S or COO. [Formula 3] In formula 3, X represents O, NH, S or COO. An anticancer composition for injection, characterized in that it contains a compound represented by the following structural formula 3. [Formula 3] In formula 3, X represents O, NH, S or COO. delete delete A drug system having an anticancer activity of an injectable formulation, comprising a compound represented by the following Structural Formula 4. [Structure 4] In Structural Formula 4, the silica nanoparticles are triethoxysilyl groups. Reacting water with 3- (triethoxysilyl) propyl isocyanate to synthesize the compound represented by the following structural formula (4). [Structure 4] In Structural Formula 4, the silica nanoparticles are triethoxysilyl groups. The anticancer composition of the injection formulation containing a compound represented by the following formula 4. [Structure 4] In Structural Formula 4, the silica nanoparticles are triethoxysilyl groups. delete