KR20200122255A - pH sensitive drug delivery system using Poly acrylic acid-graft- Polyhedral oligomeric silsesquioxane - Google Patents

Abstract

The present invention relates to: a pH-sensitive drug delivery system which contains polyacrylic acid (PAA) grafted with polyhedral oligomeric silsesquioxane (POSS) and a hydrophobic drug, and has a core-shell structure; and a pharmaceutical composition for preventing or treating cancer containing the same. The pH-sensitive drug delivery system of the present invention can encapsulate drugs with high efficiency and high dose.

Description

PH-sensitive drug delivery system using polyacrylic acid-graft polyhedral oligomeric silsesquioxane {pH sensitive drug delivery system using Polyacrylic acid-graft- Polyhedral oligomeric silsesquioxane}

The present invention relates to a pH-sensitive drug delivery system using polyacrylic acid-graft-polyhedral oligomer silsesquioxane.

The drug delivery system is to increase the tissue and cell delivery efficacy and pharmacokinetic efficacy of a compound or molecule having a pharmacological effect, and various types of drug delivery systems are currently being studied. Typically, micro/nano-level particles, liposomes, transdermal patches, inhalers, implants for drug retention, or antibody-drug conjugates are used as drug delivery systems. Particularly, micelles having a small size of 10 to 100 nm have an advantage in drug delivery because they can avoid renal filtration in the body and can avoid removal and opsonization from the reticulated endothelial system. Drugs used are mainly drugs that can act directly on proteins existing in the cytoplasm or on molecules constituting the signaling system.

Drugs acting in the cytoplasm have difficulty in passing through the cell membrane, which is a bilayer of phospholipids having amphiphilicity, so their pharmacological efficacy is inevitably largely influenced by the physico-chemical properties of the drug. In general, in the case of hydrophobic drugs, their solubility in vivo is very low, but as long as the molecules reach the extracellular matrix, they are known to spontaneously pass through the cell membrane through passive diffusion. It is focused on solving their insolubility problem within.

Korean Patent Registration No. 10-2053065

An object of the present invention is to provide a pH-sensitive drug delivery system using polyacrylic acid-graft-polyhedral oligomer silsesquioxane (PAA-graft-POSS) and a pharmaceutical composition for preventing or treating cancer comprising the same as an active ingredient.

However, the problem to be solved by the present invention is not limited to the problems mentioned above, and other problems that are not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

To achieve the above object, according to an embodiment of the present invention, polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS) and a hydrophobic drug, and having a core-shell structure, a pH-sensitive drug A carrier is provided.

In the present invention, "Polyhedral oligomeric silsesquioxane (POSS)" refers to a cage-shaped molecule (R ₈ Si ₈ O ₁₂ ) with a silicon-oxygen bond and a vertex group (R : It includes hydrogen, alkyl, alkene, aryl, arylene, etc.) and has a size of 1 to 3 nm. In the following embodiment, the polyhedral oligomeric silsesquioxane was used for 1,2-propanediol isobutyl -POSS ^® (CAS Number 480439-49-4) represented by the following general formula (1).

[Formula 1]

The polyacrylic acid (PAA) may have a molecular weight of 1000 g/mol to 5000 g/mol, more preferably 5000 g/mol, but is not limited thereto. When the molecular weight of the polyacrylic acid (PAA) polymer exceeds 5000 g/mol, agglomeration occurs severely due to strong bonds within the polymer main chain, and accordingly, the functional groups of acrylic acid do not play a role or are difficult to synthesize.

The core includes polyhedral oligomer silsesquioxane (POSS), and the shell may include polyacrylic acid (PAA) (see FIG. 1), and the hydrophobic drug is encapsulated in the core.

As described above, since the PAA present in the shell has a carboxyl group and the core contains a hydrophobic POSS, the encapsulation efficiency of the hydrophobic drug can be improved by strong interaction with the hydrophobic drug.

Here, the hydrophobic drug may be, for example, insulin, an anti-inflammatory agent, a diabetes treatment agent, a sex hormone, or an anticancer agent, but is not limited thereto.

Examples of the hydrophobic drug include insulin; Salicylates, ibuprofen, naprocene, fenoprofen, indomethacin, phenyltazone, mesotrexate, cyclophos Cyclophosphamide, mechlorethamine, dexamethasone, prednisolone, celecoxib, valdecoxib, nimesulide, cortisone and cortico Anti-inflammatory drugs such as steroids (corticoste roid); Diabetic treatments such as acetohexamide, chlorpropamide, glibenkramid, gliclazide, glipizide, torazamide, tolbutamide, pioglitazone and rosiglitazone; Sex hormones such as testosterone, progesterone, desogestrel, estrogen, estradiol, and norgestrel; Doxorubicin, Paclitaxel, cis-platin, mitomycin-C, daunomycin, 5-fluorouracil, Docetaxel, tamoxifen Tamoxifen), camptothecin, anasterozole, carbopla tin, topotecan, berotecan, irinotecan, gleevec, vincristine anticancer drugs such as vincristine); There is.

On the other hand, the term'polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS)' in the drug delivery system of the present invention refers to a graft polymer in which a large amount of polyhedral oligomer silsesquioxane is attached to a main chain made of polyacrylic acid. It means (see FIG. 1), and in the following examples, it may be expressed as'PAA-POSS','PAA-graft-POSS', and the like.

In polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS), polyhedral oligomer silsesquioxane may be grafted in a ratio of 7 to 35 per 70 acrylic acid monomers constituting the polyacrylic acid main chain, The molar ratio (PAA:POSS) of polyacrylic acid and polyhedral oligomer silsesquioxane may be in the range of 1:7 to 1:35.

The acrylic acid functional group of polyacrylic acid (PAA) serves to increase the encapsulation amount of the hydrophobic drug, and when the surrounding pH changes, the drug release can be controlled by a change in the charge of acrylic acid. However, since the size is relatively small to encapsulate the drug with only an acrylic acid functional group, the encapsulation efficiency can be further increased by adding POSS.

According to another embodiment of the present invention, including polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS) and an anticancer agent, and comprising a pH-sensitive drug delivery system having a core-shell structure as an active ingredient, A pharmaceutical composition for preventing or treating cancer is provided. As described above, the core of the drug delivery system includes polyhedral oligomer silsesquioxane (POSS), the shell includes polyacrylic acid (PAA), and an anticancer agent, a hydrophobic drug, is encapsulated in the core.

The drug delivery system contained in the pharmaceutical composition of the present invention releases an anticancer agent in the range of pH 5 to 7, which is the pH of cancer tissues having weak acidity unlike normal tissues, which is a carboxylate of polyacrylic acid chain at the pH of weakly acidic conditions. This is because the group is substituted with a proton to promote the release of the anticancer agent.

The anticancer agents include Doxorubicin, Paclitaxel, cis-platin, mitomycin-C, daunomycin, 5-fluorouracil, and docetaxel. ), Tamoxifen, Camptothecin, Anasterozole, Carbopla tin, Topotecan, Belotecan, Irinotecan, Gleevec And an anticancer agent selected from the group consisting of vincristine may be used, but the present invention is not limited thereto.

In addition, the cancer is lymphosarcoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, gastric cancer, liver cancer, rectal cancer, gallbladder cancer, bile duct cancer, colon cancer, pancreatic cancer, acute myelogenous leukemia, soft tissue osteosarcoma, breast cancer, ovarian cancer, lung cancer, bronchial cancer, bladder cancer, Wollm It may be selected from the group consisting of tumors and head and neck cancer, but is not limited thereto.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredient for administration. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, Examples include polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of the disease, the form of the drug, and the time, but can be appropriately selected by a person skilled in the art, and the daily dosage of the composition is preferably It is 0.001 mg/kg to 50 mg/kg, and it can be administered once to several times a day as needed.

According to another embodiment of the present invention,

As a method for preparing the pH-sensitive drug delivery system, 1) mixing polyacrylic acid (PAA) and polyhedral oligomer silsesquioxane (POSS) to obtain polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS). ; And

2) mixing the polyacrylic acid (PAA) grafted with the polyhedral oligomer silsesquioxane (POSS) and a hydrophobic drug to encapsulate the hydrophobic drug into the core including POSS; Comprising, there is provided a method for producing a pH-sensitive drug delivery system.

As described above, the molar ratio (PAA:POSS) of polyacrylic acid and polyhedral oligomer silsesquioxane may be 1:7 to 1:35.

In addition, as a hydrophobic drug, insulin, an anti-inflammatory agent, a diabetes treatment agent, a sex hormone, or an anticancer agent may be used, but the present invention is not limited thereto.

The pH-sensitive drug delivery system of the present invention has a hydrophobic drug encapsulation efficiency of 90% or more, and can encapsulate the drug at high efficiency and high dose.

In addition, since the drug delivery system is released in a weakly acidic environment of pH 5 to 7 in which cancer tissue is present, it may be usefully used as a pharmaceutical composition for preventing or treating cancer by encapsulating an anticancer agent.

The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be deduced from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a schematic view showing a process of manufacturing the pH-sensitive drug delivery system of the present invention.
2 is a TEM photograph of the pH-sensitive drug delivery system of the present invention.
3 shows FT-IR spectra of PAA, POSS, PAA-POSS35, PAA-POSS7, PAA-POSS35/DOX, PAA and POSS7/DOX.
Figure 4 is a comparison of the cumulative release amount of doxorubicin from the drug delivery system of the present invention at pH 5 and 7.
5 is a comparison of the viability and morphology of MDA-MB-231 cell lines treated with PAA/DOX, POSS/DOX, PAA-POSS35/DOX, PAA-POSS7/DOX, and DOX by concentration.

Hereinafter, exemplary embodiments will be described in detail with reference to the accompanying drawings. However, since various changes may be made to the embodiments, the scope of the rights of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents, or substitutes to the embodiments are included in the scope of the rights.

The terms used in the examples are used for illustrative purposes only and should not be interpreted as limiting. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present specification, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiment belongs. Terms as defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessively formal meaning unless explicitly defined in this application. Does not.

In addition, in the description with reference to the accompanying drawings, the same reference numerals are assigned to the same components regardless of the reference numerals, and redundant descriptions thereof will be omitted. In describing the embodiments, when it is determined that a detailed description of related known technologies may unnecessarily obscure the subject matter of the embodiments, the detailed description thereof will be omitted.

In addition, in describing the constituent elements of the embodiment, terms such as first, second, A, B, (a) and (b) may be used. These terms are only used to distinguish the component from other components, and the nature, order, or order of the component is not limited by the term.

Components included in one embodiment and components including common functions will be described using the same name in other embodiments. Unless otherwise stated, descriptions in one embodiment may be applied to other embodiments, and detailed descriptions in the overlapping range will be omitted.

Example One. PAA - POSS And PAA - POSS - Doxorubicin Preparation of drug delivery system

PSS-(2,3-Propanediol)propoxy-Heptaisobutyl substituted)(MW: 949.64 g/mol, Sigma Aldrich), polyacrylic acid substituted with PSS-(2,3-propanediol)propoxy-heptaisobutyl Sodium salt (poly acrylic acid partial sodium) (average MW: ~ 5000 g/mol, 50wt% in H ₂ O, Sigma Aldrich), tetrahydrofuran (MW: 72.11 g/mol, 99.5%. Samchun), sulfuric acid, doxorubicin Hydrochloride (Fluka) was prepared. Unless otherwise stated, all chemicals were used as received.

PAA 10g, PSS-(2,3-propanediol)propoxy-heptisobutyl substituted POSS (POSS substituted ratio in PAA functional group 70:7, 70:35) 95 mg and 56 ul of sulfuric acid were added to 50 ml of THF Dissolved. The mixture was stirred at 100° C. for 5 hours in a 3-neck flask using a reflux condenser. After stirring, the mixture was dried in a vacuum oven at 150° C. for 1 hour. To remove POSS that did not participate in the synthesis, the synthesized PAA-POSS was filtered with chloroform using a vacuum pump. The filtered compound was dried in a vacuum oven at 150° C. for 30 minutes to obtain a white powder.

Then, 3 ml of PAA-POSS and 1 mg of doxorubicin (DOX) were mixed in 10 ml of distilled water: THF (1:1 v/v). The solution was sonicated at 5° C. for 1 hour, stirred for 1 hour, and then dialyzed with deionized water for 6 hours using a 3.5K MWCO dialysis machine (Thermo Scientific??) to prepare PAA-POSS-DOX. Upon dialysis, PBS was changed at fixed times.

Example 2. PAA - POSS And PAA - POSS - DOX Structural characteristics

In order to confirm the structural characteristics of the PAA-POSS and PAA-POSS-DOX prepared in Example 1, observation with TEM images and infrared spectroscopy (FT-IR) were performed, respectively, and are shown in FIGS. 2 and 3. Each of 70 acrylic acid monomers in the polyacrylic acid chain was grafted into 7 and 35 POSSs, respectively, as PAA/POSS7 and PAA/POSS35.

As shown in Figure 2, PAA-POSS-DOX, a drug delivery device of the present invention, is a spherical nanoparticle having a core-shell structure, and a uniform size distribution was observed. On the other hand, the size of PAA-POSS was confirmed to be about 300±1.3 nm, indicating that doxorubicin was successfully encapsulated in the hydrophobic core.

In addition, in the FT-IR spectrum of FIG. 3, it was observed at the peak (1100 cm ^{- 1} ) indicating the stretching mode of the Si-O-Si group of POSS, and the CN stretch peak or 1260 cm ^-1 due to CO stretch vibration of the epoxide. In the POSS peak, PAA C = O group peak (1700 cm ^-1 ), PAA OH group peak (3000 cm ^-1 ) was observed (left). On the other hand, through the right graph of FIG. 3, it can be seen that the C=O peak of PAA-POSS-DOX shifted to a lower wavelength value compared to PAA-POSS, which is a strong interaction between doxorubicin and PAA-POSS. Means there is.

Example 3. Analysis of drug encapsulation efficiency

In order to measure the drug encapsulation efficiency of the drug delivery system of the present invention, the solution collected after drug loading was centrifuged (10,000 rpm, 2 hours) and the supernatant was measured absorbance at a wavelength of 480 nm with UV-VIS, thereby determining the amount of doxorubicin. Measured. The drug encapsulation efficiency of doxorubicin was calculated by the following equation (1).

Drug encapsulation efficiency (%) = total amount of drug-free drug / total amount of drug

As a result, the drug encapsulation efficiency was 67.2% and 92% in PAA and POSS-PAA-DOX containing doxorubicin, respectively, and it was confirmed that the drug encapsulation efficiency of the drug delivery system of the present invention was remarkably excellent. These results suggest that the drug delivery system of the present invention has excellent drug encapsulation ability of a hydrophobic anticancer agent.

Example 4. Doxorubicin Emission behavior

In order to analyze the release effect of doxorubicin of the drug delivery system according to the pH condition, the drug release behavior was evaluated at pH 5 and pH 7, and the results are shown in FIG. 4. The cumulative drug release amount for evaluating the doxorubicin release behavior of the drug delivery system was measured and calculated.

Referring to FIG. 4, it can be seen that the cumulative release amount at pH 5 was significantly higher compared to the pH 7 condition, and the rate of increase in the release amount was also higher at pH 5. Accordingly, the drug delivery system of the present invention may exhibit increased drug release in cancer tissues, which are weakly acidic environments.

Example 5. Cancer cells Killing effect

The cytotoxicity of the drug delivery system of the present invention prepared in Example 1 was evaluated using the human breast cancer cell line MDA-MB-231. PAA with doxorubicin (PAA/DOX), POSS with doxorubicin (POSS/DOX), PAA-POSS35 with doxorubicin (PAA-POSS35/DOX), PAA-POSS7 with doxorubicin (PAA-POSS7/DOX) ) And doxorubicin alone were treated with MDA-MB-231 cell lines at concentrations of 1, 5, and 10 μg/ml, respectively, and cell viability was measured after 1 day. The fixed MDA-MB-231 cell line was stained with crystal violet, and the colony formation state was visualized by magnifying 200 times with an AxioCam HRC CCD phase contrast microscope.

As a result, as shown in FIG. 5, PAA/DOX, POSS/DOX, PAA-POSS35/DOX, and PAA-POSS7/DOX were confirmed to exhibit cytotoxicity in a concentration-dependent manner, and in particular, PAA-POSS7/DOX 10 μg/ml When treated with, only about 65% of MDA-MD-231 cells survived, indicating the best cytotoxicity.

As described above, although the embodiments have been described by the limited drawings, a person of ordinary skill in the art can apply various technical modifications and variations based on the above. For example, the described techniques are performed in a different order from the described method, and/or components such as a system, structure, device, circuit, etc. described are combined or combined in a form different from the described method, or other components Alternatively, even if substituted or substituted by an equivalent, an appropriate result can be achieved.

Therefore, other implementations, other embodiments and claims and equivalents fall within the scope of the following claims.

Claims (15)

A polyhedral oligomer silsesquioxane (POSS) containing grafted polyacrylic acid (PAA) and a hydrophobic drug, and having a core-shell structure, a pH-sensitive drug delivery system.
The method of claim 1,
The core comprises a polyhedral oligomer silsesquioxane (POSS), and the shell comprises polyacrylic acid (PAA), a pH sensitive drug delivery system.
The method according to claim 1 or 2,
The hydrophobic drug is encapsulated in the core, a pH-sensitive drug delivery system.
The method according to claim 1 or 2,
The hydrophobic drug is insulin, an anti-inflammatory agent, a diabetes treatment agent, a sex hormone or an anticancer agent, a pH sensitive drug delivery system.
The method according to claim 1 or 2,
The molar ratio of the polyacrylic acid and the polyhedral oligomer silsesquioxane is 1:7 to 1:35, a pH-sensitive drug delivery system.
The method according to claim 1 or 2,
The drug delivery system is a pH-sensitive drug delivery system that releases a hydrophobic drug in the range of pH 5 to 7.
Polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS) and an anticancer agent, and comprising a pH-sensitive drug delivery system having a core-shell structure as an active ingredient, a pharmaceutical composition for preventing or treating cancer.
The method of claim 7,
The core includes polyhedral oligomer silsesquioxane (POSS), and the shell includes polyacrylic acid (PAA), a pharmaceutical composition for preventing or treating cancer.
The method of claim 8,
The anticancer agent is encapsulated in the core, a pharmaceutical composition for preventing or treating cancer.
The method according to claim 7 or 8,
The drug delivery system is a pharmaceutical composition for preventing or treating cancer that releases an anticancer agent in the range of pH 5 to 7.
The method according to claim 7 or 8,
The anticancer drugs doxorubicin, paclitaxel, cis-platin, mitomycin-C, daunomycin, 5-fluorouracil, docetaxel, docetaxel , Tamoxifen, camptothecin, anasterozole, carbopla tin, topotecan, berotecan, irinotecan, gleevec and A pH sensitive drug delivery system selected from the group consisting of vincristine.
The method according to claim 7 or 8,
The cancer is lymphosarcoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, gastric cancer, liver cancer, rectal cancer, gallbladder cancer, bile duct cancer, colon cancer, pancreatic cancer, acute myelogenous leukemia, soft tissue osteosarcoma, breast cancer, ovarian cancer, lung cancer, bronchial cancer, bladder cancer, and Wollm tumor. Which is selected from the group consisting of, a pharmaceutical composition for the prevention or treatment of cancer.
As a method for preparing the pH-sensitive drug delivery system of claim 1,
1) mixing polyacrylic acid (PAA) and polyhedral oligomer silsesquioxane (POSS) to obtain polyacrylic acid (PAA) grafted with polyhedral oligomer silsesquioxane (POSS); And
2) mixing the polyacrylic acid (PAA) grafted with the polyhedral oligomer silsesquioxane (POSS) and a hydrophobic drug to encapsulate the hydrophobic drug into the core including POSS; Containing, a method for producing a pH-sensitive drug delivery system.
The method of claim 13,
The molar ratio of the polyacrylic acid and the polyhedral oligomer silsesquioxane is 1:7 to 1:35, a method for producing a pH-sensitive drug delivery system.
The method of claim 13 or 14,
The hydrophobic drug is insulin, an anti-inflammatory agent, a diabetes treatment agent, a sex hormone or an anticancer agent, a method for producing a pH-sensitive drug delivery system.

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