KR100770205B1 - Drug Delivery Carrier Using Exopolysaccharide Produced from Hahella Chejuensis Mutant m10356 and Method Thereof - Google Patents

Abstract

The present invention relates to a drug carrier using the extracellular polysaccharide produced by the Hahel jeju nes mutant strain m10356 and a method for producing the same, more specifically, (a) after mixing the vegetable oil, extracellular polysaccharides and the desired drug, Stirring to form an emulsion, (b) adding acetic acid while stirring the emulsion to react, forming extracellular polysaccharide beads, and (c) centrifuging the reactants of step (b) The present invention relates to a drug carrier using an extracellular polysaccharide produced by the Haheljuju nsis mutant strain m10356, comprising recovering the drug carrier in the form of a bead, and a method for preparing the same.

The bead-type drug carrier prepared by using the extracellular polysaccharide produced by the haella jeju nsis mutant m10356 according to the present invention exhibits a sustained and excellent drug release effect at neutral pH to facilitate drug delivery to the small or large intestine.

Extracellular Polysaccharides, Drug Delivery

Description

Drug Delivery Carrier Using Exopolysaccharide Produced from Hahella Chejuensis Mutant m10356 and Method Thereof}

The present invention relates to a drug carrier using the extracellular polysaccharide produced by the Hahel jeju nes mutant strain m10356 and a method for producing the same, more specifically, (a) after mixing the vegetable oil, extracellular polysaccharides and the desired drug, Stirring to form an emulsion, (b) adding acetic acid while stirring the emulsion to react, forming extracellular polysaccharide beads, and (c) centrifuging the reactants of step (b) The present invention relates to a drug carrier using an extracellular polysaccharide produced by the Haheljuju nsis mutant strain m10356, comprising recovering the drug carrier in the form of a bead, and a method for preparing the same.

Polysaccharides secreted extracellularly by microorganisms protect microorganisms from other organisms, participate in the defense of antibodies, neutralize toxic substances in the environment, form complexes with metal ions, and evaporate intracellular moisture in dry environments. It is involved in preventing. In addition, it has been shown to perform several functions related to the survival of microorganisms (Sutherland, IW, Biotechnology , 3: 533, 1983).

Extracellular polysaccharides of microorganisms have various functions such as gel formation ability, emulsification stability, surface tension control ability, water absorption ability, adhesion ability, lubrication ability, and film formation ability depending on conditions. (Fu, JF and YH Teeng, Appl . Environ . , 56: 919, 1990). In addition, the extracellular polysaccharides have heavy metal adsorption capacity (Norberg, AG and H. Persson, Biotech . Biochem . , 115: 239, 1984) and antitumor activity (Oda, M. et. al . , Agro . Biol . Chem . , 47: 1623, 1983).

In particular, extracellular polysaccharides of microorganisms have high potential for industrial use, as described above, high productivity through improvement of production culture conditions, mass production through continuous culture using short-term fermenters, and ease of produced extracellular polysaccharides. As it has advantages such as separation and recovery, research on this is being actively conducted.

Melanin, a substance related to the skin whitening effect, absorbs ultraviolet rays to protect the skin, but when exposed for a long time, skin photoaging is promoted and pigmentation occurs. At this time, studies on materials that inhibit the activity of tyrosinase involved are actively underway. Currently, whitening products and sunscreens are mostly made of chemicals. However, the chemicals used therein may be deteriorated in biocompatibility, and may cause deterioration of skin function, acceleration of skin aging, etc., and absorb moisture in the air, resulting in skin diseases caused by adsorption of bacteria and fungi. In addition, hormonal components such as estrogen and steroids have excellent temporary effects such as skin softening effect, but may cause toxic symptoms due to commercial use.

On the other hand, angiotensin-II (angiotensin-II) is a substance produced by the conversion of angiotensin-I by an angiotensin convertin enzyme (ACE), causing a change in the locally generated blood vessel diameter and raising blood pressure, such as myocardial infarction, heart failure, etc. May cause disease. Since the disease is already complicated, the treatment method is complicated and not easy, and research for the development of ACE inhibitors and angiotensin-II antagonists, which are easy to use in the treatment, is being conducted.

On the other hand, injections used as the most common drug delivery agents have the disadvantage that they are difficult to take and painful, and should be administered periodically. In addition, the side effect is higher than oral drug delivery agent is also difficult to expect the long-term effective drug delivery efficiency.

Most of the mucous membranes in the intestine are easy to deliver drugs, rapid absorption and low molecular weight substances, as well as relatively high molecular weight substances, such as drugs with peptide structure, so that the main administration of bioactive substances Its value is very high as a route In order to use such gastrointestinal mucosa, it is urgent to develop a drug carrier that exhibits a property of inhibiting drug degeneration and release from low pH conditions in the stomach.

On the other hand, the present inventors have developed a patent for the production of extracellular polysaccharides from the haella jejuensis mutant strain m10356, which is a non-pigmented mutant strain (1010-0495272).

In the present invention, as a result of intensive efforts to identify the use of the extracellular polysaccharide prepared by the Hahel jejuensis strain m10356 (KCTC 10469 BP), it was confirmed that the extracellular polysaccharide has a bead-forming ability that can be used as a drug carrier The invention was completed.

It is an object of the present invention to provide a drug carrier using the extracellular polysaccharide produced by the Hahel jeju nsis mutant strain m10356 and a method for producing the same.

The present invention provides a method for preparing a drug carrier, comprising the following steps:

(a) 10 parts by weight of an extracellular polysaccharide (exopolysaccharide) having the following characteristics (i) to (iii) and 100 parts by weight of vegetable oil and mesalazine, sulfasalazine, ibuprofen, prednisolone, dexamethasone, budesonide, prednisolone, beclomethasone , Fluticasone, Tioxocortal, Hydrocortisone, Cyclosporin, Methotrexate, Domperidone, 5-Fluorouracil, Visacodil, Dietary Fiber, Lactic Acid Bacteria, Insulin, Vasopressin, Growth Hormone, Growth Hormone Stimulator, Colony Stimulator, Mixing a desired drug selected from the group consisting of calcitonin, immunoglobulin, diltiazem, verapamil, nifedipine, captopril, theophylline, naproxen and mixtures thereof, followed by stirring to form an emulsion;

(i) glucose, galactose and galactosamine, per week, in a molar ratio of 1: 1: 1.1;

(ii) the heterosaccharide, a subsidiary sugar, is galactronic acid; And

(iii) the molecular weight of the extracellular polysaccharide is 1.39 × 10 ⁶ Da;

(b) reacting the emulsion by adding acetic acid and then forming extracellular polysaccharide beads; And

(c) centrifuging the reactant of step (b) to recover the drug carrier in the form of a spherical bead.

In the present invention, the extracellular polysaccharide may be characterized in that it was prepared by the Hahel chejuensis mutant m10356 (KCTC 10469BP).

The present invention also provides a drug carrier which is prepared by the above method, cured at pH 6 or lower, dissolved in the range of pH 6-8, and in the form of a spherical bead having a diameter of about 150 μm.

The present invention has the effect of providing a drug delivery system and a method for producing the same using the extracellular polysaccharide produced by the Haella jeju nsis mutant strain m10356.

The bead-type drug carriers prepared using the extracellular polysaccharides produced by the Haella jeju nsis mutant m10356 according to the present invention have no side effects and exhibit a sustained and excellent drug release effect at neutral pH to the small or large intestine. Easy to drug delivery

Hereinafter, the present invention will be described in detail.

First, according to the method described in the inventors Patent No. 10-0495272, to obtain an extracellular polysaccharide (exopolysaccharide (EP)), and then to prepare the extracellular polysaccharide beads according to the method shown in Figure 1, It was confirmed that it has a high drug release effect at neutral pH.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1 Production of Extracellular Polysaccharides

According to the method described in Korean Patent Registration No. 10-0495272, the mutant strain m10356 (KCTC 10469 BP) of Hahel Jeju Ensis 95CJ10356 was prepared using M-m10356 containing sucrose, organic nitrogen, inorganic phosphoric acid, MgSO ₄ , NaCl, and trace elements. The medium was inoculated, cultured at 25 ° C. under a pH of 7, and extracellular polysaccharides were obtained from the cultured mutants.

Example 2: Preparation of Extracellular Polysaccharide Beads

30 ml of soybean oil was added to two tubes, 3 ml of 1.0% extracellular polysaccharide was added to one side, and 3 ml of 1.5% extracellular polysaccharide was added to the other side. Each of the two tubes containing soybean oil and extracellular polysaccharides was added to 0.3 ml of ovalbumin to form a W / O emulsion with vigorous stirring. 15 ml of acetic acid was added while the stirring was continued while the W / O emulsion was reacted for 2 hours. The beads are recovered by centrifugation, washed with diethyl ether, dried at 50 ° C. for 2 hours, and lyophilized to finally obtain extracellular polysaccharide beads (drug carriers). It was. Ovalbumin (OVA) was used to observe the drug release effect.

Figure 1 schematically shows a method for producing extracellular polysaccharide beads according to the present invention. Figure 2 is a photograph showing the shape of the extracellular polysaccharide beads prepared by the above method, it was confirmed that the extracellular polysaccharide beads of the present invention having a spherical particle size of about 150μm.

Example 3: Drug Release Effect of Extracellular Polysaccharide Beads

Two tubes containing 50 ml of PBS buffer (pH 7.4) were prepared, and two types of extrapolysaccharide beads (prepared with 1.0% and 1.5% extracellular polysaccharides) prepared in Example 5 were prepared, respectively. Put in. In addition, the mixture according to the above procedure was prepared using HCl-KCl buffer (pH 1.4) instead of PBS buffer.

While incubating the reaction at 37 ° C, a portion of the buffer was extracted at regular intervals and absorbance was measured at 590 nm. Using the measured absorbance, the concentration of ovalbumin released from the extracellular polysaccharide beads was measured according to a previously prepared calibration curve.

As a result, as shown in FIG. 3, in the case of beads prepared using 1.0% extracellular polysaccharide, the release of ovalmine was not observed until 12 hours of culture under the condition of pH 1.4, and about 20 hours after 48 hours of culture. It showed a release effect of%, and under the condition of pH 7.4, it showed a release effect of about 55% at 4 hours of cultivation, and then gradually increased to about 98.2% at 48 hours of cultivation. .

In addition, in the case of beads prepared using 1.5% extracellular polysaccharide, drug release proceeded more slowly than beads prepared using 1.0% extracellular polysaccharide, which showed about 50% release effect after 8 hours of culture. Afterwards, it gradually increased and showed a release effect of about 94.54% after 48 hours of culture.

For both types of beads, release at pH 7.4 was about 70 times higher than release at pH 1.4 at 12 hours of incubation.

In addition, through the above experimental results, the extracellular polysaccharide beads of the present invention exhibits a hardening property at low pH, while the pH-dependent drug release tends to dissolve at neutral pH, which is an enteric condition. It was confirmed that the formulation is very suitable for drug release using gastrointestinal mucosa.

Having described the specific part of the present invention in detail, it will be 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.

Figure 1 schematically shows a method for producing extracellular polysaccharide beads according to the present invention.

Figure 2 shows the form of the extracellular polysaccharide beads according to the present invention.

Figure 3 shows the drug release effect of the extracellular polysaccharide beads according to the present invention.

Claims (3)

A method for preparing a drug carrier comprising the following steps: (a) 10 parts by weight of an extracellular polysaccharide (exopolysaccharide) having the following characteristics (i) to (iii) and 100 parts by weight of vegetable oil and mesalazine, sulfasalazine, ibuprofen, prednisolone, dexamethasone, budesonide, prednisolone, beclomethasone , Fluticasone, thioxocortal, hydrocortisone, cyclosporin, methotrexate, domperidone, 5-fluorouracil, bisacodil, dietary fiber, lactic acid bacteria, insulin, vasopressin, growth hormone, growth hormone stimulating factor, colony stimulating factor, Mixing a desired drug selected from the group consisting of calcitonin, immunoglobulin, diltiazem, verapamil, nifedipine, captopril, theophylline, naproxen and mixtures thereof, followed by stirring to form an emulsion; (i) glucose, galactose and galactosamine, per week, in a molar ratio of 1: 1: 1.1; (ii) the heterosaccharide, a subsidiary sugar, is galactronic acid; And (iii) the molecular weight of the extracellular polysaccharide is 1.39 × 10 ⁶ Da; (b) reacting the emulsion by adding acetic acid and then forming extracellular polysaccharide beads; And (c) centrifuging the reactant of step (b) to recover the drug carrier in the form of a spherical bead. The method of claim 1, wherein the extracellular polysaccharide is manufactured by a Hahella chejuensis strain m10356 (KCTC 10469BP). A drug carrier in the form of a spherical bead prepared by the method of claim 1 or 2, cured at pH 6 or less, dissolved in the range of pH 6-8, and having a diameter of about 150 μm.

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