KR20250019597A - Novel microspheres using anionic polymer, preparation method and composition thereof - Google Patents

Abstract

The present invention relates to a novel microsphere using an anionic polymer, a manufacturing method, etc., and the present invention provides a novel manufacturing method and an optimal formulation of microspheres which are useful for the delivery of cationic drugs, have low toxicity, are biocompatible, and can deliver drugs to microvessels, and the novel manufacturing method according to the present invention improves the yield of microspheres of a desired size by controlling the size distribution of microsphere particles, thereby enabling industrial mass production of microspheres of a specific size.

Description

{NOVEL MICROSPHERES USING ANIONIC POLYMER, PREPARATION METHOD AND COMPOSITION THEREOF}

The present invention relates to novel microspheres and a manufacturing method using anionic polymers.

Liver cancer is a malignant tumor with a poor prognosis, ranking second in Korea and third worldwide in terms of cause of death due to cancer. More than 70% of liver cancer patients cannot undergo radical surgery, and even in cases where radical resection is performed, the probability of recurrence in other parts of the body within 5 years is more than 50%. The possibility of responding to systemic chemotherapy in advanced liver cancer is very low, less than 10%, and various image-guided therapies have been developed as alternative non-surgical treatment methods. There are two representative image-guided therapies applied to liver cancer: transarterial chemoembolization (TACE), which locally injects anticancer drugs into the liver cancer through the hepatic artery and concurrently performs embolization of the hepatic artery, and radiofrequency ablation (RFA).

More than 95% of the blood flow to liver cancer tissue is delivered to the hepatic artery, but more than 75% of the blood flow to surrounding healthy tissue is delivered to the portal vein and only 25% is delivered to the hepatic artery, so TACE is a safe and effective treatment method that does not cause damage to normal tissue.

TACE is usually performed repeatedly 3 to 4 times a year, so the administered substance must be safe. The drugs mainly used in existing arterial embolization include doxorubicin, cisplatin, epirubicin, mitoxantrone, and mitomycin C. Since these anticancer drugs cannot be directly dissolved in water-insoluble contrast media, they are dissolved in water-soluble contrast media such as Pamiray and then homogeneously dispersed in water-insoluble contrast media (iodinated oil) such as Lipiodol in the form of an oil-in-water or water-in-oil emulsion. Since this emulsion undergoes phase separation after a certain period of time, it is prepared and used immediately before patient administration, and chemical embolization is performed and additionally gelatin sponge particles are administered to promote ischemic necrosis of cancer tissue and prevent drug loss. However, the administered emulsion undergoes phase separation in a short period of time after administration, and the anticancer drug is absorbed at once, so the antitumor effect is not continuous, and side effects due to systemic exposure may occur. In addition, most cytotoxic anticancer drugs administered in TACE are metabolized in the liver, so they may exhibit hepatotoxicity. For this reason, patients show postembolization syndrome, and daily life is difficult, so hospitalization for 2-3 days is inevitable, and medical costs are high.

Recently, drug-eluting beads (DEBs) that are selectively used are made of non-absorbable polyvinyl alcohol, etc., and are administered by adsorbing drugs to the embolic particles through ion exchange. The particle size ranges from 40 to 900 μm. Compared to emulsion droplets, the particle range is homogeneous and reproducible, and the drug is slowly released, so there is an advantage of less systemic exposure to the drug. However, since the drug is not completely released after administration, less than 50% is released by 28 days after administration, and about 90% is released after 90 days, so the drug accumulates in high concentrations in cancer tissues relatively slowly. In addition, since it takes more than 1 to 2 hours to adsorb the drug, and since it is non-absorbable, the particles remain in the body even after the drug is completely released, which can cause necrosis of normal tissues, making it difficult to repeat TACE, and there is a risk of cancer recurrence. In addition, the cost of the drug is high compared to the existing TACE method, so the financial burden on the patient is large.

Therefore, there is a need for the development of microspheres that are low in toxicity, biocompatible, and can deliver target drugs to microvessels quickly and efficiently.

The Korean Journal of Hepatology 2010 ; 16:242-246

The present inventors have made extensive efforts to develop biocompatible microspheres that are easy to manufacture and have excellent drug-loading capacity, and as a result, have identified a formulation optimized for the manufacture of microspheres, and have confirmed that the yield of particles of a desired size can be improved by controlling the size distribution of microsphere particles by controlling the rpm of a homogenizer, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a composition for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt as effective ingredients.

Another object of the present invention is to provide a method for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt.

Another object of the present invention is to provide a method for preparing a composition for treating cancer.

Another object of the present invention is to provide microspheres comprising an anionic polymer and a polyvalent metal compound salt.

However, the technical problems to be achieved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

To achieve the above purpose, the present invention provides a composition for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt as effective ingredients.

In addition, the present invention provides a use for producing microspheres of an anionic polymer and a polyvalent metal compound salt; or a composition comprising the same as an active ingredient.

In addition, the present invention provides a use of an anionic polymer and a polyvalent metal compound salt for the production of microspheres; or a composition comprising the same as an active ingredient.

In one embodiment of the present invention, the anionic polymer may be selected from the group consisting of, but is not limited to, carboxymethyl cellulose or a salt thereof; dextran sulfate or a salt thereof; and mixtures thereof.

In another embodiment of the present invention, the microspheres may include, but are not limited to, carboxymethylcellulose and dextran sulfate.

In another embodiment of the present invention, the carboxymethylcellulose and dextran sulfate may be included in a mass ratio of, but not limited to, 0.5 to 20:1.

In another embodiment of the present invention, the carboxymethylcellulose and dextran sulfate may be included in a concentration ratio of, but not limited to, 0.5 to 20:1.

In another embodiment of the present invention, the polyvalent metal compound salt may be, but is not limited to, one or more divalent to tetravalent metal compound salts selected from the group consisting of Zr ²⁺ , Zr ⁴⁺ , Fe ²⁺ , Fe ³⁺ , Ca ²⁺ , Al ³⁺ , Cu ²⁺ , and Zn ²⁺ .

In another embodiment of the present invention, the anionic polymer and the polyvalent metal compound salt may be included in a concentration ratio of 0.1 to 10:1, but is not limited thereto.

In another embodiment of the present invention, the microspheres may be loaded with a cationic drug, but are not limited thereto.

In another embodiment of the present invention, the cationic drug may be at least one selected from the group consisting of doxorubicin, procainamide, digoxin, quinidine, trimethoprim, cimetidine, vancomycin, irinotecan, daunorubicin, epirubicin, diphenhydramine, memantine, oxycodone, pyrilamine, tramadol, and pharmaceutically acceptable salts thereof, but is not limited thereto.

In another embodiment of the present invention, the cationic drug may be loaded into the microspheres at a mass ratio of cationic drug: microsphere = 1:1 to 10, but is not limited thereto.

In another embodiment of the present invention, the composition may be, but is not limited to, an emulsion formulation.

In another embodiment of the present invention, the microspheres may have a diameter of, but not limited to, 10 to 700 μm.

In another embodiment of the present invention, the microspheres may have a size distribution as follows, but is not limited thereto:

a) 10 to 50 μm;

b) 50 to 100 μm;

c) 100 to 300 μm;

d) 300 to 500 μm; and

e) 500 to 700 μm.

In another embodiment of the present invention, the microspheres can be separated according to their size distribution, but are not limited thereto.

In another embodiment of the present invention, the microspheres can be used for, but are not limited to, cancer treatment.

In another embodiment of the present invention, the microspheres can be used in, but are not limited to, vascular embolization.

In addition, the present invention provides a method for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt, comprising the following steps:

A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;

A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution; and

A step of separating microspheres of a desired size using a sieve.

In one embodiment of the present invention, the anionic polymer may be selected from the group consisting of, but is not limited to, carboxymethyl cellulose or a salt thereof; dextran sulfate or a salt thereof; and mixtures thereof.

In another embodiment of the present invention, the polyvalent metal compound salt may be, but is not limited to, one or more divalent to tetravalent metal compound salts selected from the group consisting of Zr ²⁺ , Zr ⁴⁺ , Fe ²⁺ , Fe ³⁺ , Ca ²⁺ , Al ³⁺ , Cu ²⁺ , and Zn ²⁺ .

In another embodiment of the present invention, the nonionic surfactant may be selected from the group consisting of, but is not limited to, polyvinyl alcohol, polyalkylene glycol, polyvinyl pyrrolidone, alkyl polyglycoside, t-octylphenoxy polyethoxy ethanol, poloxamer, polysorbate, poly(N-vinylacetamide), polyvinyl butyral, sorbitan oleate, and mixtures thereof.

In another embodiment of the present invention, the oil phase may be selected from the group consisting of, but not limited to, mineral oil, vegetable oil, medium chain triglycerides, and mixtures thereof.

In another embodiment of the present invention, the water phase and the oil phase may be mixed in a volume ratio of 1:1 to 20, but is not limited thereto.

In another embodiment of the present invention, the emulsion may be added to the polyvalent metal compound salt aqueous solution in a volume ratio of 1:1 to 20, but is not limited thereto.

In another embodiment of the present invention, the method can control the size distribution of microspheres by controlling the rpm of the homogenizer, but is not limited thereto.

The present invention also provides a method for preparing a composition for treating cancer, comprising the following steps:

A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;

A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution;

A step of separating microspheres of a desired size using a sieve; and

A step of mixing a cationic drug with the microspheres.

Additionally, the present invention provides microspheres comprising an anionic polymer and a polyvalent metal compound salt.

Additionally, the present invention provides a drug delivery application of the microspheres.

The present invention also provides the use of said microspheres for the manufacture of drug delivery vehicles.

In addition, the present invention provides a composition for cationic drug delivery comprising the microspheres and a cationic drug as active ingredients.

Furthermore, the present invention provides a cationic drug delivery method comprising the step of administering the microspheres encapsulated with a cationic drug to a subject in need thereof.

In addition, the present invention provides a composition comprising the microspheres and a cationic drug as active ingredients for use in treating vascular embolism and/or cancer.

In addition, the present invention provides a composition for treating cancer comprising the microspheres and a cationic drug as active ingredients.

In addition, the present invention provides a method for treating vascular embolism and cancer, comprising a step of administering to a subject a composition comprising the microspheres and a cationic drug as active ingredients.

In addition, the present invention provides a use of a composition comprising the microspheres and a cationic drug as active ingredients for the manufacture of a vascular embolization and cancer treatment agent.

The present invention provides a novel method for producing microspheres, which are useful for the delivery of cationic drugs, have low toxicity, are biocompatible, and can deliver drugs to microvessels, and an optimal formulation. In addition, the method for producing microspheres of the present invention controls the particle size according to the size of blood vessels by controlling the size distribution of microsphere particles, thereby enabling the microspheres to directly embolize blood vessels, and improves the yield of microspheres of a desired size, thereby enabling mass production of microspheres of a specific size.

Figure 1 shows an image of the change in the reaction solution when an anionic polymer and calcium chloride (CaCl ₂ ) react.
Figure 2 shows an optical microscope measurement image of a solid produced through Composition 2 of Example 1-2, <Reactivity Test 2>.
Figure 3 shows an image of microparticles manufactured through the reaction of anionic polymer and iron chloride according to Composition 2 of Example 3, Table 3.
Figure 4 shows the results of measuring the particle distribution of microspheres manufactured using a bead mill using dynamic light scattering (DLS).
Figure 5 shows a TEM measurement image of microspheres manufactured using a bead mill.
Figure 6 shows a drug dissolution graph after loading a cationic drug (doxorubicin hydrochloride).
Figure 7 shows the results of measuring cell survival rate in Huh7 cells.
Figure 8 shows an optical microscope image of an emulsion of a cationic drug-loaded microsphere suspension solution and Lipiodol Ultra solution.
Figure 9 shows the microcatheter passage images of each suspension using 5% sucrose and 15% mannitol as drug-loaded aqueous suspensions and a mixed emulsion with Lipiodol Ultra solution.
Figure 10 shows the appearance of microspheres manufactured in Example 8.
Figure 11 shows the appearance of doxorubicin-bound microspheres manufactured in Example 8.
Figure 12 shows the dissociation over time of the microspheres manufactured in Example 8.
Figure 13 shows the dissociation over time of doxorubicin-bound microspheres manufactured in Example 8.
Figure 14 shows the synthesis results of CMC-DS microspheres according to the type of crosslinking agent.
Figure 15 shows the binding of doxorubicin (AD mycin) to FeCl ₃ or ZrOCl ₂ cross-linked CMC-DS microspheres.
Figure 16 shows CMC-DS microspheres with controlled size distribution.
Figure 17 shows microsphere particles formed depending on the concentration of cross-linking agent.
Figure 18 shows the results of confirming the stability of the W/O emulsion used in the production of microspheres.
Figures 19a and 19b show the results of observing microspheres manufactured under the conditions of 5% CMC and 0% DS under a microscope. Figure 19a shows microspheres with sizes of 25 to 50 and 50 to 100 um, and Figure 19b shows microspheres with sizes of 100 to 300 and 300 to 500 um.
FIGS. 20A to 20E show the results of observing microspheres with different CMC:DS ratios of 100 to 300 and 300 to 500 μm in size under a microscope. FIG. 20A shows CMC:DS=5:1 microspheres, FIG. 20B shows CMC:DS=4:1 microspheres, FIG. 20C shows CMC:DS=3:1 microspheres, FIG. 20D shows CMC:DS=2:1 microspheres, and FIG. 20E shows CMC:DS=1:1 microspheres.
Figure 21a shows the results of examining the difference in properties before and after drug loading to confirm the drug loading ability of microspheres according to the size of the microspheres and the drug:microsphere ratio.
Figure 21b shows the results of checking the properties after drug loading to confirm the drug loading ability of microspheres according to the CMC:DS ratio.

The microspheres according to the present invention are composed of carboxymethyl cellulose, which is an anionic polymer of the cellulose series, and dextran series with biodegradable properties, and are characterized by using a cross-linking agent to increase aqueous stability, so that they have low toxicity and are biocompatible, and can not only deliver drugs to microvessels, but also obtain microspheres of a desired size, so that the size of the microspheres can be adjusted according to the size of the blood vessel so that the microspheres can directly embolize the blood vessel, and microspheres of a desired size can be provided.

Accordingly, the present invention provides a composition for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt as effective ingredients.

In addition, the present invention provides microspheres comprising an anionic polymer and a divalent metal compound salt, manufactured using the composition for manufacturing microspheres, and according to one embodiment of the present invention, the microspheres may include an anionic polymer and a divalent metal compound salt, but are not limited thereto.

In the present invention, the anionic polymer may be selected from the group consisting of carboxymethyl cellulose or a salt thereof; dextran sulfate or a salt thereof; and mixtures thereof, but is not limited thereto.

In the present invention, the microspheres may include, but are not limited to, carboxymethyl cellulose and dextran sulfate.

In another embodiment of the present invention, the carboxymethylcellulose and dextran sulfate are mixed in a ratio of 0.5 to 20:1, 0.5 to 15:1, 0.5 to 10:1, 0.5 to 5:1, 0.5 to 4:1, 0.5 to 3:1, 0.5 to 2:1, 0.5 to 1:1, 1 to 3:1, 1.5 to 2.5:1, 1 to 5:1, 2 to 5:1, 3 to 5:1, 3.5 to 4.5:1, 4 to 5:1, 4:1, 1 to 10:1, 1 to 9:1, 3 to 9:1, 4 to 9:1, 5 to 9:1, 6 to 9 : may be included in a mass ratio of, but is not limited to, 1, 7 to 9: 1, 8 to 9: 1, 7.5 to 8.5: 1, or 8: 1.

In the present invention, the carboxymethyl cellulose and dextran sulfate are mixed in an amount of 0.5 to 20:1, 0.5 to 15:1, 0.5 to 10:1, 0.5 to 5:1, 0.5 to 4:1, 0.5 to 3:1, 0.5 to 2:1, 0.5 to 1:1, 1 to 3:1, 1.5 to 2.5:1, 1 to 5:1, 2 to 5:1, 3 to 5:1, 3.5 to 4.5:1, 4 to 5:1, 4:1, 1 to 10:1, 1 to 9:1, 3 to 9:1, 4 to 9:1, 5 to 9:1, 6 to 9:1, It may be included in a concentration ratio (% w/w) of 7 to 9: 1, 8 to 9: 1, 7.5 to 8.5: 1, or 8: 1, and according to one embodiment of the present invention, the carboxymethyl cellulose and dextran sulfate may be included in a concentration ratio (% w/w) of 2 to 5: 1, preferably, but not limited to, a concentration ratio (% w/w) of 4: 1.

In the present invention, the carboxymethyl cellulose and dextran sulfate may be included in a concentration ratio of preferably 1 to 5:1, 1 to 4:1, 1 to 3:1, 1 to 2:1, 2 to 5:1, 2 to 4:1, 2 to 3:1, 5:1, 4:1, 3:1, 2:1, 1:1, or 1:0, but is not limited thereto.

In the present invention, the polyvalent metal compound salt may be at least one divalent to tetravalent metal compound salt selected from the group consisting of Zr ²⁺ , Zr ⁴⁺ , Fe ²⁺ , Fe ³⁺ , Ca ²⁺ , Al ³⁺ , Cu ²⁺ , and Zn ²⁺ , and specifically, the polyvalent metal compound salt may be iron (II) chloride, iron (III) chloride, aluminum chloride, iron nitrate [Fe(NO ₃ ) ₃ ], aluminum nitrate [Al(NO ₃ ) ₃ ], iron acetate [Fe(CH ₃ COO) ₃ ], aluminum acetate [Al(CH ₃ COO) ₃ ], iron perchlorate [Fe(ClO ₄ ) ₃ ], zinc chloride [ZnCl ₂ ], calcium chloride [CaCl ₂ ], zirconium oxychloride [ZrOCl ₂ ·8H ₂ O], and copper(II) nitrate trihydrate [Cu(NO ₃ ) ₂ ·3H ₂ O]. According to one embodiment of the present invention, the multivalent metal compound salt may be zirconium oxychloride [ZrOCl ₂ ·8H ₂ O], or iron(III) chloride, but is not limited thereto.

According to one embodiment of the present invention, the anionic polymer is carboxymethyl cellulose or a salt thereof, dextran sulfate or a salt thereof; and the polyvalent metal compound salt may be zirconium oxychloride [ZrOCl ₂ 8H ₂ O], iron (III) chloride, but is not limited thereto.

In the present invention, the anionic polymer and the polyvalent metal compound salt may be included in a concentration ratio (% w/w) of 0.1 to 10:1, 0.3 to 10:1, 0.5 to 10:1, 0.6 to 10:1, 0. 8 to 10:1, 1 to 10:1, 3 to 10:1, 5 to 10:1, 8 to 10:1, 0.5 to 8:1, 0.5 to 5:1, 0.5 to 3:1, 0.5 to 2.5:1, 0.6 to 5:1, 0.6 to 3:1, 0.6 to 2.5:1, 0.6 to 1.5:1, or 0. 6 to 1:1, and according to one embodiment of the present invention, It may be included in a concentration ratio of 0.6 to 2.4: 1, preferably 0.6: 1, but is not limited thereto.

In the present invention, the microspheres may be loaded with a cationic drug, and the cationic drug may be loaded in a form combined with anions present inside and on the surface of the microspheres. That is, the microspheres according to the present invention can deliver cationic drugs into the body as a drug delivery vehicle.

The cationic drug may be at least one selected from the group consisting of doxorubicin, procainamide, digoxin, quinidine, trimethoprim, cimetidine, vancomycin, irinotecan, daunorubicin, epirubicin, diphenhydramine, memantine, oxycodone, pyrilamine, tramadol, and pharmaceutically acceptable salts thereof, and according to one embodiment of the present invention, may be doxorubicin, but is not limited thereto.

In the present invention, the cationic drug may be loaded into the microspheres at a mass ratio of cationic drug: microsphere = 1:1 to 10, 1:1 to 9, 1:1 to 8, 1:1 to 7, 1:1 to 6, 1:1 to 5, 1:1 to 4, 1:1 to 3, 1:1 to 2.5, 1:1 to 1.5, or 1:1, but is not limited thereto.

In the present invention, the cationic drug and microspheres may be included in a dry weight ratio of, but not limited to, 1:1 to 10, 1:1 to 9, 1:1 to 8, 1:1 to 7, 1:1 to 6, 1:1 to 5, 1:1 to 4, 1:1 to 3, 1:1 to 2.5, 1:1 to 1.5, or 1:1.

In the present invention, the microspheres may include carboxymethyl cellulose and dextran sulfate, wherein the carboxymethyl cellulose and dextran sulfate are mixed in an amount of 0.5 to 20:1, 0.5 to 15:1, 0.5 to 10:1, 0.5 to 5:1, 0.5 to 4:1, 0.5 to 3:1, 0.5 to 2:1, 0.5 to 1:1, 1 to 3:1, 1.5 to 2.5:1, 1 to 5:1, 2 to 5:1, 3 to 5:1, 3.5 to 4.5:1, 4 to 5:1, 4:1, 1 to 10:1, 1 to 9:1, 3 to 9: It may be included in a mass ratio or concentration ratio (% w/w) of 1, 4 to 9:1, 5 to 9:1, 6 to 9:1, 7 to 9:1, 8 to 9:1, 7.5 to 8.5:1, or 8:1. In addition, in the present invention, the anionic polymer and the polyvalent metal compound salt may be included in a concentration ratio (% w/w) of 0.1 to 10:1, 0.3 to 10:1, 0.5 to 10:1, 0.6 to 10:1, 0. 8 to 10:1, 1 to 10:1, 3 to 10:1, 5 to 10:1, 8 to 10:1, 0.5 to 8:1, 0.5 to 5:1, 0.5 to 3:1, 0.5 to 2.5:1, 0.6 to 5:1, 0.6 to 3:1, 0.6 to 2.5:1, 0.6 to 1.5:1, or 0. 6 to 1, and loaded into the microsphere. The cationic drug may be loaded at a mass ratio of microspheres to cationic drug: microspheres = 1:1 to 10, 1:1 to 9, 1:1 to 8, 1:1 to 7, 1:1 to 6, 1:1 to 5, 1:1 to 4, 1:1 to 3, 1:1 to 2.5, 1:1 to 1.5, or 1:1, but is not limited thereto.

In the present invention, the composition may be an emulsion formulation, and the emulsion formulation in the present invention may be prepared by mixing an aqueous phase to which an anionic polymer and a nonionic surfactant are added and an oil phase to which a nonionic surfactant is added using a homogenizer, but is not limited thereto.

In the present invention, the microspheres have a size of 10 to 700 μm, 10 to 600 μm, 10 to 500 μm, 10 to 400 μm, 10 to 300 μm, 10 to 200 μm, 10 to 100 μm, 20 to 700 μm, 20 to 600 μm, 20 to 500 μm, 20 to 400 μm, 20 to 300 μm, 20 to 200 μm, 20 to 100 μm, 20 to 50 μm, 25 to 700 μm, 25 to 600 μm, 25 to 500 μm, 25 to 400 μm, 25 to 300 μm, 25 to 200 μm, 25 to It may have a diameter of, but is not limited to, 100 μm, or 25 to 50 μm.

In the present invention, the microspheres have the following size distribution, so that the microspheres according to the present invention can be classified (separated) according to the following diameter range, i.e., the size distribution, and microspheres of a specific size can be separated and used according to the intended use:

a) 10 to 50 μm, preferably 25 to 50 μm;

b) 50 to 100 μm;

c) 100 to 300 μm;

d) 300 to 500 μm; and

e) 500 to 700 μm, preferably 500 μm or more.

In the present invention, the microspheres can be used as a drug delivery vehicle capable of delivering an anticancer agent into the body, and can also block blood vessels directly to block blood flow, so they can be used for cancer treatment and/or vascular embolization, but are not limited thereto. Since the microspheres of the present invention can be separated according to particle size, vascular embolization can be performed by separating microspheres suitable for the size of the blood vessels at the site where vascular embolization is to be performed.

Accordingly, the present invention can provide a pharmaceutical composition for treating cancer comprising the microspheres and a cationic drug as active ingredients, or a composition for delivering a cationic drug comprising the microspheres and a cationic drug as active ingredients.

The above-mentioned vascular embolization is transarterial chemoembolization (TACE), which can be a method of treating cancer by injecting an anticancer drug into an artery leading to cancer tissue and using an embolic substance to block the supply of nutrients to the cancer tissue. In addition, the above-mentioned vascular embolization can be a method of treating a patient suffering from excessive bleeding, vascular malformation, or tumor by injecting a substance into a blood vessel to stop blood flow to a specific blood vessel, thereby blocking blood flow through the blood vessel.

In the present invention, the cancer includes angiogenesis-dependent cancer, preferably at least one selected from the group consisting of liver cancer, ovarian cancer, breast cancer, and non-small cell lung cancer, and more preferably liver cancer.

In the present invention, when a cationic drug is combined with the microspheres and then mixed with iodinated oil, which is an embolic agent, a stable emulsion can be formed, and the embolic effect can be increased by injecting the emulsion into the hepatic artery. Therefore, the composition of the present invention may further include an embolic agent, and the embolic agent may be iodinated oil, but is not limited thereto.

The above iodized oil may be at least one selected from the group consisting of iodized oil derived from poppy seeds, iodized oil derived from soybeans, and ethiodol, but is not limited thereto. The above iodized oil derived from poppy seeds may use the product name Lipiodol, but is not limited thereto.

The composition of the present invention is characterized in that it is administered within 0 to 2 hours, preferably within 0 to 1 hour, after preparation, and has excellent stability since the movement of the drug within the microspheres is maintained low for a long period of time compared to compositions used in conventional chemoembolization.

In the present invention, the composition including the microspheres and the cationic drug and the embolic material can be mixed using, but is not limited to, a mixing method using a 3-way cock, a method using ultrasound, etc.

In addition, the present invention provides a method for preventing or treating vascular embolism and cancer, comprising a step of administering the composition to a subject.

The content of the microspheres and cationic anticancer agent in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., and may be, for example, 0.0001 to 99.9 wt% or 0.001 to 50 wt% based on the total weight of the composition, but is not limited thereto. The content ratio is a value based on the dry amount after removing the solvent.

Additionally, the present invention provides microspheres comprising an anionic polymer and a polyvalent metal compound salt.

The present invention also provides a drug delivery application of the microspheres.

The present invention also provides the use of said microspheres for the manufacture of drug delivery vehicles.

In addition, the present invention provides a composition for cationic drug delivery comprising the microspheres and a cationic drug as active ingredients.

Furthermore, the present invention provides a cationic drug delivery method comprising the step of administering the microspheres encapsulated with a cationic drug to a subject in need thereof.

In addition, the present invention provides a composition comprising the microspheres and a cationic drug as active ingredients for use in treating vascular embolism and/or cancer.

In addition, the present invention provides a composition for treating cancer comprising the microspheres and a cationic drug as active ingredients.

In addition, the present invention provides a method for treating vascular embolism and cancer, comprising a step of administering to a subject a composition comprising the microspheres and a cationic drug as active ingredients.

In addition, the present invention provides a use of a composition comprising the microspheres and a cationic drug as active ingredients for the manufacture of a vascular embolization and cancer treatment agent.

The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. The excipients may be, for example, at least one selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film-coating materials, and controlled-release additives.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, 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.

When formulating, it is usually prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

Additives that can be used in the liquid formulation according to the present invention include water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, etc.

The syrup according to the present invention may use a solution of white sugar, other sugars or sweeteners, and may also use a fragrance, a colorant, a preservative, a stabilizer, a suspending agent, an emulsifier, a viscosity increasing agent, etc., as needed.

Purified water may be used in the emulsion according to the present invention, and an emulsifier, preservative, stabilizer, fragrance, etc. may be used as needed.

The suspension according to the present invention may use suspending agents such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose, HPMC 1828, HPMC 2906, and HPMC 2910, and may also use surfactants, preservatives, stabilizers, colorants, and fragrances as needed.

The injection according to the present invention includes: solvents such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, nonvolatile oils such as sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethyl acetamide, butazolidine, propylene glycol, Tween, nitertidine amide, hexamine, and dimethyl acetamide; buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and gums; It may include isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO ₃ ), carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), and ethylenediaminetetraacetic acid; sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid disodium, and acetone sodium bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium cisplatinum, sodium alginate, Tween 80, and aluminum monostearate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, the "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the excretion rate, the treatment period, the concurrently used drugs, and other factors well known in the medical field.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art to which the present invention belongs.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with various related factors such as the disease to be treated, route of administration, age, sex, weight of the patient, and severity of the disease.

In the present invention, the term "subject" means a subject requiring treatment of a disease, and more specifically, may be a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, or cow, but is not limited thereto.

In the present invention, “administration” means providing a composition of the present invention to a subject by any appropriate method.

In the present invention, “prevention” means any act of suppressing or delaying the onset of a target disease, “treatment” means any act of improving or beneficially changing a target disease and its resulting metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and “improvement” means any act of reducing a parameter related to a target disease, for example, the degree of symptoms, by administering a composition according to the present invention.

In addition, the present invention provides a kit for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt. The kit may further include instructions describing a method for producing the microspheres of the present invention.

In addition, the present invention provides a drug delivery kit comprising the microspheres; and/or a cancer prevention or treatment kit comprising microspheres encapsulating an anticancer agent.

In the present invention, the "kit" is not limited in its specific form or type, and a kit of a form commonly used in the art can be used, and in addition to the microspheres of the present invention, any component necessary for carrying out the purpose of the present invention can be further included. For example, in addition to the microspheres of the present invention, a storage solution, an administration tool, a drug, a suspension for administration, etc. can be additionally included, and an instruction manual containing information regarding the microspheres of the present invention can be additionally included.

In addition, the present invention provides a method for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt, comprising the following steps:

(S1) A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;

(S2) a step of adding the above emulsion to a multivalent metal compound salt aqueous solution to form microspheres; and

(S3) A step of separating microspheres of a desired size using a sieve.

Matters relating to the above composition can be applied to the present manufacturing method.

The above step (S1) may include, but is not limited to, one or more selected from the group consisting of steps a) to c) below.

The above step (S2) may include, but is not limited to, steps d) and/or e) below.

The above step (S3) may include, but is not limited to, the following step f).

The method for manufacturing the above microspheres may include, but is not limited to, a step of stirring between each step so that each component is uniformly mixed.

In the present invention, the emulsion may be in a W/O form, but is not limited thereto.

The above (W/O) emulsion may have a (volume) ratio of the water phase to the oil phase of, but is not limited to, 1:1 to 20, 1:1 to 15, 1:1 to 10, 1:1 to 8, 1:1 to 5, 1:1 to 4, 1:1 to 3, 1:1 to 2, 1: 3 to 15, 1:3 to 10, 1:3 to 8, 1:3 to 5, 1:3 to 4, 1:4 to 10, 1:4 to 8, 1:4 to 5, 1:20, 1:15, 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, or 1:1. In the present invention, the higher the ratio of the oil phase, the better the emulsion formation can be achieved, but the optimal ratio for using the oil phase to a minimum may be preferably 1:4.

In the present invention, in the step (S2), the emulsion may be added to the polyvalent metal compound salt aqueous solution at a volume ratio of 1:1 to 20, 1:1 to 15, 1:1 to 10, 1:1 to 7.5, 1:1 to 5, 1:1 to 2.5, 1:1 to 2, 1:2 to 15, 1:2 to 10, 1:2 to 7.5, 1:2 to 5, 1:6 to 10, 1:6 to 9, 1:6 to 7.5, 1:7.5 to 9, 1:20, 1:15, 1:10, 1:9, 1:8, 1:7.5, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, or 1:1, and according to one embodiment of the present invention, It can be added in a volume ratio of 1:7.5, but is not limited thereto.

According to one embodiment of the present invention, it was confirmed that the size distribution of the microspheres produced can be controlled by controlling the rpm of the homogenizer that mixes the water phase and the oil phase in the step (S1). Therefore, the method for producing microspheres of the present invention is characterized in that the particle distribution of the microspheres can be controlled by controlling the rpm of the homogenizer.

According to one embodiment of the present invention, the microspheres separated using the 500, 300, 100, 53, and/or 25 μm sieves in the step (S3) may have a size distribution as follows, but is not limited thereto, and the size distribution may change depending on the type of sieve used.

a) 10 to 50 μm, preferably 25 to 50 μm;

b) 50 to 100 μm;

c) 100 to 300 μm;

d) 300 to 500 μm; and

e) 500 to 700 μm, preferably 500 μm or more.

In the present invention, the method may further include a step of purifying and washing the microspheres separated using a sieve, but is not limited thereto.

In the present invention, purification of the microspheres can be performed using a centrifuge or a filter, but is not limited thereto.

In the present invention, the washing may be selected from the same organic solvent or the group of solvents mentioned above, but is not limited thereto.

In the present invention, the washing may be performed using, after the organic solvent washing, an aqueous solvent such as distilled water, water for injection, sucrose, mannitol, or sodium chloride dissolved therein, and a solvent selected from the group consisting of these, but is not limited thereto.

The method for producing microspheres of the present invention may specifically include one or more of the following steps, or may consist of the following steps, but is not limited thereto:

a) a step of dissolving one or more anionic polymers and nonionic surfactants in the water;

b) a step of dissolving a nonionic surfactant in the oil phase;

c) a step of forming an emulsion by stirring the water phase of a) and the oil phase of b) using a homogenizer;

d) a step of forming microspheres by adding the emulsion of step c) to a crosslinking agent aqueous solution (a solution of a polyvalent metal compound salt) in which a crosslinking agent (a salt of a polyvalent metal compound) is dissolved in an aqueous ethanol solution;

e) Additional stirring step in the above step d),

f) a step of separating microspheres according to particle size using a 500, 300, 100, 53, and/or 25 μm sieve in the solution of step e);

g) a step of separating and/or removing residual organic solvent from the microspheres using an aqueous solution;

h) Step of removing moisture by placing it in the oven.

The above steps a) to c) may be performed in any suitable order.

Additionally, the above steps are not necessarily limited to the order described above.

In the present invention, the water may be deionized water or distilled water, but is not limited thereto.

In the present invention, the oil phase may be selected from the group consisting of mineral oil, vegetable oil, medium chain triglyceride (MCT), and mixtures thereof, and according to one embodiment of the present invention, the oil phase may be medium chain triglyceride, but is not limited thereto.

In the present invention, the mineral oil is also called mineral oil, and is an oil whose main components are alkane and paraffin, which is a by-product produced in the process of refining crude oil.

In the present invention, the vegetable oil may be an oil derived from nuts, seeds, grains, etc., and may include, for example, peanut oil, soybean oil, coconut oil, or olive oil.

In the present invention, the medium-chain triglyceride is a representative alternative fat manufactured by hydrolyzing palm oil and palm oil, fractionating caprylic acid and capric acid, etc., and then esterifying (or combining) this with glycerol. It is a medium-chain fatty acid with a small molecular structure and usually 8 to 10 carbon atoms.

In the present invention, the nonionic surfactant may be selected from the group consisting of polyvinyl alcohol, polyalkylene glycol, polyvinyl pyrrolidone, alkyl polyglycoside, t-octylphenoxy polyethoxy ethanol, poloxamer, polysorbate (preferably polysorbate 20 (Tween 20)), poly(N-vinylacetamide), polyvinyl butyral, sorbitan oleate (Span 80), and mixtures thereof, and according to one embodiment of the present invention, the nonionic surfactant may be selected from the group consisting of polysorbate 20 and/or sorbitan. It may be sorbitan oleate, and preferably, the nonionic surfactant dissolved in the water phase together with the anionic polymer may be polysorbate 20, and the nonionic surfactant dissolved in the oil phase may be sorbitan oleate, but is not limited thereto.

In the present invention, the polyalkylene glycol may be any of the conventional polyalkylene glycol series known in the art without limitation, but it is preferable to use a polyethylene glycol series.

In the present invention, the poloxamer is a nonionic triblock copolymer in which hydrophobic propylene oxide is bonded to hydrophilic ethylene oxide at both ends, and has the characteristic of being temperature sensitive, so that it can be converted between sol and gel depending on the concentration and temperature, and the properties of the poloxamer change depending on the ratio of polyoxypropylene and polyoxyethylene. The above poloxamer is poloxamer 101, poloxamer 105, poloxamer 105 benzoate, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 182 dibenzoate, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, may be selected from the group consisting of, but is not limited to, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, and poloxamer 407.

In the present invention, the alkyl polyglycoside may be selected from the group consisting of, for example _{, C 8-22 alkyl} glycosides, more specifically, caprylyl glucoside, coco-glucoside, decyl glucoside, lauryl glucoside, methyl coco glucoside, and myristyl glucoside, but is not limited thereto.

In the present invention, the organic solvent may be at least one selected from the group consisting of alkyl alcohols, ketones, acetonitrile, tetrahydrofuran, and mixtures thereof, but is not limited thereto.

The above alkyl alcohols may be alcohols having 1 to 6 carbon atoms, and examples thereof include, but are not limited to, methanol, ethanol, alcohol, propanol, butanol, pentanol, and hexanol.

The above ketones may be organic solvents having the chemical formula of RCOR, and may be, but are not limited to, acetone, methyl ethyl ketone, methyl butyl ketone, cyclohexanone, methyl isobutyl ketone, diisobutyl ketone, etc.

In addition, the present invention provides a method for preparing a composition for treating vascular embolism and cancer, comprising the following steps:

A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;

A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution;

A step of separating microspheres of a desired size using a sieve; and

A step of mixing a cationic drug with the microspheres.

The step of mixing the cationic drug with the microspheres may be performed by, but is not limited to, ultrasonic treatment.

The method for preparing the composition for vascular embolism and cancer treatment may further include, but is not limited to, a step of mixing the mixed composition of the microspheres and the cationic drug with an embolic substance.

The mixing step with the above-mentioned color material can be performed using, but is not limited to, a mixing method using a 3-way cock, a method using ultrasonic waves, etc.

The term "ultrasound" used in the present invention generally means sound waves exceeding the frequency of 16 Hz to 20 kHz, which is the sound wave frequency that the human ear can hear, and high-intensity focused ultrasound introduces focused ultrasound that provides continuous and high-intensity ultrasonic energy to a focus, and can produce instantaneous thermal effects (65-100℃), cavitation effects, mechanical effects, and sonochemical effects depending on the energy and frequency. Ultrasound is not harmful when passing through human tissue, but high-intensity ultrasound that forms a focus generates sufficient energy to cause coagulation necrosis and thermocautery effects regardless of the type of tissue.

In the present invention, the ultrasonic waves refer to sound waves having a frequency higher than the audible frequency range of 16 Hz to 20 kHz.

The present invention also provides a method for preparing a cationic drug delivery composition comprising the following steps:

A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;

A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution;

A step of separating microspheres of a desired size using a sieve; and

A step of mixing a cationic drug with the microspheres.

In this specification, “active ingredient” means an ingredient that exhibits the intended activity alone or can exhibit the intended activity together with a carrier or the like that is inactive on its own.

Throughout the specification of the present invention, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components can be included, unless otherwise specifically stated. The terms "about," "substantially," and the like, used throughout the specification of the present invention, are used in a meaning at or close to the numerical value when manufacturing and material tolerances inherent in the stated meaning are presented, and are used to prevent unscrupulous infringers from unfairly utilizing the disclosure in which exact or absolute values are mentioned to aid the understanding of the present invention.

Throughout the specification of the present invention, the term "a combination of these" included in the expressions in the Makushi format means a mixture or combination of one or more selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

The use of the term "above" and similar referential terms in this specification (especially in the claims) may refer to both the singular and the plural. In addition, when a range is described, it is intended to include individual values within the range (unless otherwise stated), and each individual value constituting the range is described in the detailed description. Finally, unless the order of the steps constituting the method is explicitly stated or otherwise stated to the contrary, the steps may be performed in any suitable order. The order in which the steps are described is not necessarily limited. The use of all examples or exemplary terms (e.g., etc.) is intended merely to further illustrate the technical idea and is not intended to limit the scope of the invention by reason of the examples or exemplary terms, unless otherwise stated by the claims. Furthermore, those skilled in the art will recognize that various modifications, combinations, and variations may be made within the scope of the appended claims or their equivalents, depending on design conditions and factors.

The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are only used to distinguish one component from another. For example, without departing from the scope of the present invention, the first component may be referred to as the second component, and similarly, the second component may also be referred to as the first component. The term and/or includes any combination of a plurality of related described items or any item among a plurality of related described items.

The present invention can be modified in various ways and has various embodiments, and specific embodiments are illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to specific embodiments, and it should be understood that it includes all modifications, equivalents, or substitutes included in the spirit and technical scope of the present invention. In describing the present invention, if it is judged that a specific description of a related known technology may obscure the gist of the present invention, the detailed description thereof will be omitted.

Hereinafter, preferred examples are presented to help understand the present invention. However, the following examples are provided only to help understand the present invention more easily, and the content of the present invention is not limited by the following examples.

[Example]

Example 1. Test to confirm the reactivity of polymer and crosslinking agent

To compare the reactivity between anionic polymers and calcium chloride, the reactivity was tested by adding calcium chloride in the presence of CMC (Carboxymethyl cellulose).

Example 1-1. Reactivity Test 1

2 mL of 0.2% CMC and 2 mL of 0.2% carboxymethyl dextran sodium (hereinafter referred to as CMD) were added to a 20 mL container, and 0.5 mL of 10% calcium chloride was added while stirring. No solids were formed after checking for 1 hour.

Example 1-2. Reactivity Test 2

The reactivity was confirmed by conducting a test in the same manner as <Reactivity Test 1> for each composition in Table 1 below by increasing the concentration.

As shown in Figure 1, there was no change in the case of composition 1, and compositions 2 and 3 were changed into suspensions.

In addition, as shown in Fig. 2, the optical microscope analysis image of the reaction solid of composition 2 of Table 1 confirmed that it was gelled and not granulated.

According to the above results, it was confirmed that calcium chloride is not suitable as a cross-linking agent for producing nanoparticles.

Reactivity Test 2 Composition number CMC CMD CaCl ₂ Microparticles
Whether formed or not Composition 1 - 20% 5% With a clear solution
Not formed Composition 2 7% - 5% Microscopic examination of suspension showed no particle formation and gelation Composition 3 7% 20% 5% Microscopic examination of suspension showed no particle formation and gelation

Example 2. Manufacturing of microparticles using W/O emulsion

According to the composition ratios in Table 2 below, CMC, iron chloride (FeCl ₃ ) solution, and DS (dextran sulfate) powder were prepared, respectively.

First, DS powder was dissolved in CMC solution, mixed with 1% polyvinyl alcohol (PVA) solution at room temperature, and then added dropwise to medium chain triglyceride (MCT) oil used as the oil phase, and an emulsion was prepared using a sonicator. FeCl ₃ was added dropwise to the emulsion, and further stirred to prepare particles. To obtain the formed particles, centrifugation was performed at 4°C and 15,000 rpm for 10 minutes, and insoluble matters and MCT oil were removed using ethanol. Ethanol and purified water were sequentially separated using a centrifuge under the same conditions, and finally redispersed in 2 mL of purified water.

Test composition and conditions number CMC DS PVA Sonication
time W/O
Ratio FeCl ₃ Bridge time Composition 1 0.50%, 0.5mL 10mg 1%, 0.5mL 90s 1: 20 0.50% 2hr Composition 2 0.50%, 0.5mL 10mg 1%, 0.5mL 90s 1:5 0.40% 2hr Composition 3 0.50%, 0.5mL 10mg 1%, 0.5mL 180s 1:20 0.40% 2hr Composition 4 0.10%, 0.5mL 10mg 1%, 0.5mL 90s 1:40 0.40% 2hr Composition 5 0.10%, 0.5mL 10mg 1%, 0.5mL 90s 1:20 0.40% 2hr Composition 6 0.50%, 0.5mL 10mg 1%, 0.5mL 90s 1:40 0.40% 2hr

As a result, particle formation was improved as the CMC concentration increased and the sonication time increased.

Example 3. Confirmation of yield by manufacturing microparticles using W/O emulsion

For scale-up, a homogenizer was used instead of an ultrasonic disperser. Using a homogenizer (rpm: 17,000), particles were manufactured at room temperature according to the composition ratios in Table 3 below using the same manufacturing method, and the yields for each composition ratio were compared. Finally, each redispersed suspension was freeze-dried and the mass was measured after drying.

As shown in Fig. 3, through the image of the solid produced through composition 2 of Table 3, it was found that particles were formed when iron chloride was used rather than when calcium chloride was used.

As shown in Table 3, the highest yield was obtained when CMC and DS were mixed at a mass ratio of 2:1.

Test composition, conditions, yields, and yields number CMC DS PVA
(mass%) Homogenization
hour W/O
Ratio Crosslinking agent
(mass%) Bridge
hour Amount of gain transference number Composition 1 1%, 100mL,
(1g) 2g 2%, 100mL 10min 1: 4 2.5% FeCl ₃ , 100mL 30min 1.08g 36.0% Composition 2 1%, 100mL,(1g) 0.5g 2%, 100mL 10min 1: 4 2.5% FeCl ₃ , 100mL 30min 0.83g 79.0% Composition 3 1%, 100mL,(1g) 1g 2%, 100mL 10min 1: 4 2.5% FeCl ₃ , 100mL 30min 1.01g 50.5% Composition 4 2%, 100mL, (2g) 0.5g 2%, 100mL 10min 1:4 2% AlCl ₃ , 100mL 30min 1.23g 49%

(However, the yield excludes the cross-linking agent content)

Example 4. Manufacturing of microspheres through additional nanoprocessing after manufacturing microparticles using W/O emulsion

The microparticles were manufactured using the following method. To manufacture the aqueous solution, 2 g of PVA and 1 g of CMC were dissolved in 100 mL of distilled water, respectively. Then, 0.5 g of DS was measured, added to the CMC solution, and dissolved. To manufacture the crosslinking agent solution, 0.5 g of iron (III) chloride (FeCl ₃ ) was dissolved in 100 mL of distilled water.

To prepare a W/O emulsion, 800 mL of MCT oil was charged into a 2 L beaker, and all of the water phase solution was added while stirring at 17,000 rpm at room temperature using a homogenizer. After stirring for 10 minutes under the same conditions, the cross-linking agent solution was added dropwise and stirred for an additional 30 minutes.

The following experimental method was used to separate the microparticles: The reaction solution was transferred to a separatory funnel and allowed to stand for 2 hours. The lower layer (aqueous phase) containing yellow solids in the separatory funnel was collected, and a centrifuge was used to generate a yellow particle pellet. The supernatant was discarded, and the white material was dissolved and removed using ethanol. At this time, care was taken not to affect the lower layer pellet. 15 mL of ethanol was added, and the pellet was redispersed using a vortex and an ultrasonic disperser, and this was repeated until no lumps were visible. Then, the washing process was sequentially repeated with distilled water and a sucrose (5%) solution in addition to ethanol, to obtain microparticles. Finally, it was redispersed in a 5% sucrose solution.

A bead mill (Minicer, Netzsch) was used to produce microspheres. The washed particles were diluted in a 5% sucrose solution to produce microspheres in an amount of about 500 mL. The particles were made small by milling at a pump speed of 20 rpm, a flow rate of 5 m/s, and about 10°C using 0.2 mm beads in the bead mill equipment. After replacing the beads with 0.1 mm, additional milling was performed at a pump speed of 10 rpm, a flow rate of 4 m/s, and the same temperature.

As shown in Fig. 4, the DLS (dynamic light scattering) measurement results showed that the particle distribution was 50 nm to 460 nm.

In addition, as shown in Fig. 5, it was confirmed that the particles were well formed into a spherical shape.

Example 5. Preparation and dissolution test of microspheres loaded with doxorubicin hydrochloride

Doxorubicin hydrochloride was completely dissolved in a 5% sucrose solution at a concentration of 25 mg/mL. The completely dissolved doxorubicin hydrochloride sucrose solution was added to 1.25 mL of a 5% sucrose solution in which 60 mg of microspheres were dispersed to adjust the ratio of doxorubicin hydrochloride : particle = 25 mg : 60 mg, and after an initial 1-minute sonication (bath type sonication), the mixture was additionally mixed for approximately 30 minutes.

500 μL of the microsphere suspension loaded with the above doxorubicin hydrochloride was added to the Cellu Sep membrane (MW: 15,000, Width: 32 mm, Wall thickness: 24 μm), and both sides were closed with clips. 50 mL of buffer (PVA 0.4%, Tween 0.1% in lactic acid pH=2.7, 37 degrees) was placed in a vial, stirred at 200 rpm, and the temperature was increased to 37 degrees. The membrane sample was added, and a dissolution test was performed. The absorbance of the samples taken at each point was measured using a UV spectrometer (wavelength: 475 nm) to confirm the dissolution amount.

As a result, as shown in Fig. 6, the doxorubicin release rate was found to be excellent, at approximately 70%.

Example 6. In vitro Cytotoxicity assessment

To evaluate the toxicity of the manufactured microspheres, a viability assay was performed in Huh7 liver cancer cells. Three groups were evaluated: doxorubicin hydrochloride (dox), particles loaded with doxorubicin (032d), and particles not loaded with doxorubicin (032).

As shown in Figure 7, it was confirmed that particles loaded with the same drug as doxorubicin hydrochloride itself exhibited a cancer cell killing effect in a concentration-dependent manner, whereas particles not loaded with the drug showed no killing effect regardless of concentration, indicating no toxicity.

Example 7. Preparation of mixed emulsion with Lipiodol and confirmation of passage through microcatheter

In a vial containing 50 mg of doxorubicin hydrochloride (Ildong Adriamycin RDF), 1.25 mL of Omnipaq 300 (GE Healthcare) was added to dissolve the drug. The entire amount was taken up using a syringe, and the solution was added to a solution containing 60 mg of microspheres dispersed in 2 mL of a 5% sucrose solution to prepare a suspension in which the drug was loaded onto nanoparticles. In addition, 10 mL of Lipiodol Ultra solution (Guerbet Korea) was prepared in a separate syringe. The two solutions were placed in a 3-way cock, and the syringes were pushed alternately to prepare an emulsion. An image of the formed emulsion is shown in Fig. 8.

A microcatheter was connected to the last end of the three-way cock containing the above-mentioned manufactured emulsion, and the syringe was pushed to pass through it. In the same manner, a microcatheter passage test was performed to compare the microcatheter passage test with an emulsion manufactured by replacing the 5% sucrose solution with a 15% mannitol solution.

As a result, as shown in Fig. 9, in both cases, the microcatheter was passed through.

Therefore, it was confirmed that the emulsion prepared by the method of the present invention can be used for embolization by passing through a microcatheter.

Example 8. Preparation of microspheres (I) by combining CMC (Carboxymethylcellulose), DS (dextran sulfate), and doxorubicin

CMC-DS microspheres were prepared by an ionic cross-linking method. First, 5 g of CMC and 1.25 g of DS were dissolved in 95 g of deionized water. The CMC and DS solutions were stirred at room temperature for 2 hours. As a cross-linking agent, 2.5 g of zirconium oxychloride octahydrate (ZrOCl ₂ 8H ₂ O) was completely dissolved in 100 g of deionized water (DI water) at room temperature. The CMC and DS mixture was slowly added dropwise to the ZrOCl ₂ 8H ₂ O solution. The CMC and DS microspheres were cross-linked for 2 hours and washed with deionized water. The appearance of the CMC and DS microparticles was analyzed under a microscope. As a result, transparent microspheres were observed, as shown in Fig. 10. To bind doxorubicin hydrochloride to the microspheres, a doxorubicin hydrochloride solution (Drug product) was mixed with the microspheres (N=13) at a concentration of 2 mg/mL and stored overnight at room temperature. The appearance of the doxorubicin-bound microspheres was observed under a microscope. As a result, as shown in Fig. 11, it was confirmed that the doxorubicin-bound microspheres turned red.

Example 9. Dissociation of microspheres and doxorubicin binding microspheres

Empty microspheres and doxorubicin-conjugated microspheres were dissociated in PBS (phosphate buffered saline, pH 7.4). The microspheres were each stored in a vial containing 100 mL of deionized water, and the vials were stored at room temperature while observing the dissociation of the microspheres for 7 days.

As a result, as shown in FIGS. 12 and 13, it was confirmed that both empty microspheres and doxorubicin-bound microspheres dissociated over time.

Example 10. Synthesis of CMC-DS microspheres dependent on cross-linking agent

CMC-DS microspheres were synthesized using various cross-linking agents. The method for preparing the microspheres followed the method of Example 8, except for the cross-linking agent solution. All cross-linking agent experiments were performed independently.

First, 5 g of CMC and 5 g of DS were dissolved in 100 mL of deionized water. 1 g of ZnCl ₂ , 0.834 g (0.5%) of FeCl ₂ , 0.834 g (0.5%) of FeCl ₃ , 2.5 g of CaCl ₂ , and 2.5 g of ZrOCl ₂ ·8H ₂ O were each dissolved in 100 mL of deionized water. As cross-linkers, ZnCl ₂ , FeCl _{2 ,} FeCl ₃ , CaCl ₂ , and ZrOCl ₂ ·8H ₂ O were respectively applied to the CMC-DS microspheres. The CMC-DS mixture was added dropwise to each cross-linker and stirred for 2 hours. After cross-linking, the CMC-DS microspheres were washed with deionized water. The appearance of each CMC-DS microsphere was observed under a microscope.

As a result, as shown in Fig. 14, FeCl ₃ and ZrOCl ₂ 8H ₂ O were effectively bound to DS through electrostatic bonding. On the other hand, ZnCl ₂ , FeCl _{2 , and} CaCl ₂ were ineffective in forming the CMC-DS microsphere surface, so that the CMC-DS microspheres were not maintained and were separated or all merged.

Example 11. FeCl ₃ or ZrOCl ₂ Doxorubicin binding to cross-linked CMC-DS microspheres

For AD mycin (doxorubicin hydrochloride) binding test to CMC-DS microspheres, FeCl ₃ or ZrOCl ₂ cross-linked CMC-DS microspheres were evaluated. 1 mL of AD mycin (2 mg/mL) was added to each microsphere and incubated for 6 h. AD mycin binding was confirmed by the change in appearance color.

As shown in Figure 15, both the FeCl ₃ cross-linked CMC-DS microspheres and the ZrOCl ₂ cross-linked CMC-DS microspheres bound AD mycin, and it was confirmed that more than 90% of AD mycin was loaded into the microspheres.

Example 12. Size distribution control

CMC-DS microspheres were fabricated by controlling the size distribution. The fabrication of CMC-DS microspheres for controlling the size distribution was accomplished by emulsification and filtration methods. First, 2% CMC and 0.5% DS were dissolved in water. The CMC and DS mixture was added dropwise to MCT oil containing 4% SPAN 80. The ratio of the aqueous phase (W) to the oil phase (O) was 25%, and the mixture was homogenized at 3000 rpm for 10 min. The CMC-DS emulsion (W/O) was further added dropwise to 2.5% ZrOCl ₂ 8H ₂ O solution, a cross-linking agent, at a rate of 3 mL/min. The cross-linking procedure was accomplished by stirring at 200 rpm. The CMC-DS microspheres were sequentially filtered through sieves with pore sizes of 500 μm, 300 μm, 100 μm, and 53 μm. The optical visualization of the microspheres filtered through each sieve is shown in Fig. 16.

Example 13. Optimization of the formulation of microspheres

In the process of optimizing the formulation of microspheres, the raw materials used were as shown in Table 4, the devices used were as shown in Table 5, and the analytical instruments used were as shown in Table 6.

Raw material name Manufacturing place Mixing purpose Sodium carboxymethyl cellulose (CMC-Na) Ashland chief ingredient Sodium dextran sulfate (DS-Na) BOC Sci. chief ingredient Medium Chain Triglycerides (MCT) KLK-OLEO Emulsion composition Tween 20 Thermo scientific Emulsion additive Span 80 Sigma-Aldrich Emulsion additive Zirconyl chloride octahydrate Sigma-Aldrich Crosslinking agent

Manufacturing process Manufacturing equipment equipment name Manufacturing/Sales Place W/O emulsion manufacturing Homogenizer Silverson Bridge Mechanical stirrer LKlabkorea Bridge Orbital shaker Allforlab

equipment name Equipment Number Manufacturer model name Balance RE-21-017 METTLER TOLEDO XPR105DRV UV-vis spectrophotometer RE-16-012 SHIMADZU UV-1800

Example 13-1. ZrOCl ₂ Optimization of crosslinker concentration

As shown in Table 7, the aqueous phase was added dropwise to the ZrOCl ₂ cross-linking agent at concentrations of 2.5, 5, and 10% (w/w%) using a syringe (26G syringe needle) to manufacture microspheres. When the aqueous phase was added dropwise, the cross-linking agent was stirred using an orbital shaker at 150 rpm. The appearance of the manufactured microspheres was visually confirmed to confirm the appropriate cross-linking agent concentration.

As a result, as shown in Fig. 17, when the CMC/DS concentrations were each 5%, it was determined that a cross-linking agent concentration of 10% (w/w%) or higher was suitable for particle formation.

Example 13-2. Stability of W/O emulsion

For the homogeneous production of microspheres, the stability of the W/O emulsion can be an important factor, and the stability of the emulsion can be controlled by a surfactant.

Therefore, in order to secure an emulsion formulation suitable for microsphere production (W/O/W), the stability of the emulsion was confirmed according to the addition of surfactants Span 80 and Twen 20 under the conditions of Table 8.

The HLB (Hydrophile-Lipophile Balance) of condition 1 was calculated as follows:

HLB = 16.7(Tween 20 HLB) x 0.1 + 4.3(Span 80 HLB) x 0.1 = 2.1

Theoretically, it is known that a low HLB (2.0 to 6.0) is suitable for W/O emulsions. As a result of observing the degree of emulsion stabilization with the naked eye and a microscope, as shown in Fig. 18, the experimental results also confirmed that the emulsion under condition 1 was more stable than the emulsion under condition 2, which is consistent with the theory.

Example 13-3. Confirmation of the possibility of manufacturing microspheres without DS

Award (W): 5% CMC, 0% DS, 10% Tween 20 added to distilled water

O: 10% Span 80 added to MCT oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 300 rpm to prepare a cross-linking agent solution. 0.5 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

As a result of observing the manufactured microspheres under a microscope, as shown in FIGS. 19a and 19b, it was shown that when particles were manufactured through W/O emulsion manufacturing compared to the dropping method, it was easy to manufacture spherical microspheres by micro-sized emulsion manufactured by a homogenizer.

Therefore, it was confirmed that the above method for manufacturing microspheres can control the size of the microspheres manufactured according to the size of the W emulsion of the W/O emulsion by controlling the stirring speed (RPM) of the homogenizer.

Example 13-4. Production of microspheres according to CMC:DS ratio

<CMC:DS = 5:1 Microsphere Manufacturing>

W: 5% CMC, 1% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

<CMC:DS = 4:1 Microsphere Manufacturing>

W: 4.8% CMC, 1.2% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

<CMC:DS = 3:1 Microsphere Manufacturing>

W: 4.5% CMC, 1.5% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

<CMC:DS = 2:1 Microsphere Manufacturing>

W: 4.0% CMC, 2.0% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

<CMC:DS = 1:1 Microsphere Manufacturing>

W: 3.0% CMC, 3.0% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then particles were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 um. Each particle obtained by size was placed in an oven at 60 degrees to remove moisture.

As a result of observing microspheres manufactured at different CMC:DS ratios under a microscope, as shown in FIGS. 20a to 20e, when the CMC:DS ratio was 1:1, no microspheres were formed. Therefore, it was determined that the CMC:DS ratio should be 2:1 or higher for easy formation of microspheres.

Example 13-5. Drug loading capacity according to CMC:DS ratio

The CMC:DS ratio microspheres manufactured in Example 13-4 were classified by size (100-300 μm). The classified microspheres were placed in a 60 degree oven to completely remove moisture, then weighed and re-injected with distilled water to adjust the concentration to 1 mg/mL. 0.25, 0.5, 1, and 2 mg of doxorubicin drug were dissolved in 2 mL of distilled water, respectively. 2 mg/2 mL of microspheres and 0.25, 0.5, 1, or 2 mg/2 mL of drug were mixed, and then vortexed for 30 seconds to mix. After 7 days of drug loading, 50 uL of the supernatant was sampled, diluted in 1950 uL of distilled water, and the drug loading rate was measured by measuring UV-vis absorbance, and the drug loading ability was calculated.

The results of examining the properties before and after drug loading according to the size of the microspheres based on CMC:DS=5:1 are shown in Fig. 21a, the drug loading amount according to the CMC:DS ratio is shown in Table 9, and the results of examining the properties after drug loading according to the CMC:DS ratio are shown in Fig. 21b.

Example 14. Optimization of the manufacturing process of microspheres

Example 14-1. Confirmation of microsphere size distribution according to W/O emulsion conditions

The size distribution of microspheres manufactured according to the homogenizer conditions of W/O emulsion was confirmed. Specifically, the size distribution of microspheres was confirmed as follows.

W: 5% CMC, 1% DS, 10% Tween 20 added to distilled water

O: Add 10% Span 80 to MCT Oil

The above-mentioned water (W) and oil (O) phases were each mixed in a W:O = 30 mL:120 mL ratio using a Homogenizer at 1000 rpm or 2000 rpm for 10 minutes to prepare a W/O emulsion. 30 mL of a solution containing 10% ZrOCl ₂ 8H ₂ O added to 90% EtOH aqueous solution was placed in a 100 mL beaker and stirred using a mechanical stirrer at 180 rpm to prepare a cross-linking agent solution. 4.0 mL of the prepared W/O emulsion was pipetted into the cross-linking agent solution, and then microspheres were obtained and washed by size using sieves of 500, 300, 100, 53, and 25 μm. Each microsphere obtained by size was placed in a 60 degree oven to remove moisture, and then the weight of the microsphere by size was measured to confirm the size distribution.

As a result, as shown in Tables 10 and 11, it was confirmed that a large amount of microspheres of 100 to 500 μm in size were produced under the 1000 rpm condition, and a large amount of microspheres of 25 to 100 μm in size were produced under the 2000 rpm condition. From this, it was found that the size of the microspheres was controlled depending on the rpm condition of the Homogenizer of the W/O emulsion. The weight measurement results of the microspheres by size depending on the rpm of the Homogenizer are shown in Tables 10 and 11.

Homogenizer operating conditions (1000 rpm, 10 minutes)

Homogenizer operating conditions (2000 rpm, 10 minutes)

Example 14-2. Confirmation of the yield of microspheres according to the CMC:DS ratio

As shown in Table 12, after manufacturing the microspheres, the weight of the manufactured microspheres was measured to confirm the yield of microspheres according to the CMC:DS ratio.

As a result, as shown in Table 13, when 4 mL of W/O emulsion was injected for crosslinking under each condition, an average of 70.2 mg of microspheres could be obtained, and the deviation in the amount of microspheres obtained under each condition was confirmed to be 4.74 mg.

The yield of microspheres according to the CMC:DS ratio is shown in Table 13.

As described above, the novel microspheres using the anionic polymer according to the present invention were manufactured by injecting a W/O emulsion containing CMC and DS into a crosslinking agent aqueous solution, and the manufactured microspheres exhibited size distributions of 25-50, 50-100, 100-300, and 300-500 um. In addition, in order to find the optimal formulation for manufacturing the microspheres of the present invention, the CMC/DS ratio was divided into 5:1, 4:1, 3:1, 2:1, and 1:1 to manufacture microspheres. As a result, it was confirmed that the CMC/DS ratio of 2:1 or higher is necessary for the easy manufacture of microspheres, which are spherical particles. In addition, as a result of loading the cationic drug Doxorubicin onto the microspheres and confirming the loading ability according to the CMC/DS ratio, it was confirmed that 0.3771, 0.5611, 0.5112, 0.4488, and 0.3490 mg of drug were loaded per 1 mg of particles for 5:1, 4:1, 3:1, 2:1, and 1:1, respectively. Therefore, it was determined that the CMC/DS ratio of 4:1 is optimal in terms of the ease of manufacturing microspheres and the drug loading ability.

Furthermore, it was confirmed that the particle distribution of the microspheres can be controlled by controlling the rpm conditions of the homogenizer used in the production of the W/O emulsion in the production process of the microspheres of the present invention.

The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential characteristics of the present invention. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

Claims (26)

A composition for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt as active ingredients.
In the first paragraph,
A composition, characterized in that the anionic polymer is selected from the group consisting of carboxymethyl cellulose or a salt thereof; dextran sulfate or a salt thereof; and mixtures thereof.
In the first paragraph,
A composition characterized in that the above microspheres comprise carboxymethylcellulose and dextran sulfate.
In the third paragraph,
A composition characterized in that the carboxymethyl cellulose and dextran sulfate are included in a mass ratio of 0.5 to 20:1.
In the third paragraph,
A composition characterized in that the carboxymethyl cellulose and dextran sulfate are included in a concentration ratio of 0.5 to 20:1.
In the first paragraph,
A composition characterized in that the above polyvalent metal compound salt is at least one divalent to tetravalent metal compound salt selected from the group consisting of Zr ²⁺ , Zr ⁴⁺ , Fe ²⁺ , Fe ³⁺ , Ca ²⁺ , Al ³⁺ , Cu ²⁺ , and Zn ²⁺ .
In the first paragraph,
A composition characterized in that the anionic polymer and the polyvalent metal compound salt are included in a concentration ratio of 0.1 to 10:1.
In the first paragraph,
A composition characterized in that the microspheres are loaded with a cationic drug.
In Article 8,
A composition characterized in that the cationic drug is at least one selected from the group consisting of doxorubicin, procainamide, digoxin, quinidine, trimethoprim, cimetidine, vancomycin, irinotecan, daunorubicin, epirubicin, diphenhydramine, memantine, oxycodone, pyrilamine, tramadol, and pharmaceutically acceptable salts thereof.
In Article 8,
A composition characterized in that the cationic drug is loaded into the microspheres at a mass ratio of 1:1 to 10.
In the first paragraph,
A composition, characterized in that the composition is in an emulsion formulation.
In the first paragraph,
A composition, characterized in that the microspheres have a diameter of 10 to 700 μm.
In the first paragraph,
A composition characterized in that the above microspheres have a size distribution as follows:
a) 10 to 50 μm;
b) 50 to 100 μm;
c) 100 to 300 μm;
d) 300 to 500 μm; and
e) 500 to 700 μm.
In Article 13,
A composition characterized in that the above microspheres are separated according to size distribution.
In the first paragraph,
A composition characterized in that the above microspheres are used for cancer treatment.
In the first paragraph,
A composition characterized in that the above microspheres are used in vascular embolization.
A method for producing microspheres comprising an anionic polymer and a polyvalent metal compound salt, comprising the following steps:
A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;
A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution; and
A step of separating microspheres of a desired size using a sieve.
In Article 17,
A method, characterized in that the anionic polymer is selected from the group consisting of carboxymethyl cellulose or a salt thereof; dextran sulfate or a salt thereof; and mixtures thereof.
In Article 17,
A method characterized in that the nonionic surfactant is selected from the group consisting of polyvinyl alcohol, polyalkylene glycol, polyvinyl pyrrolidone, alkyl polyglycoside, t-octylphenoxy polyethoxy ethanol, poloxamer, polysorbate, poly(N-vinylacetamide), polyvinyl butyral, sorbitan oleate, and mixtures thereof.
In Article 17,
A method, characterized in that the oil phase is selected from the group consisting of mineral oil, vegetable oil, medium chain triglycerides, and mixtures thereof.
In Article 17,
A method characterized in that the above-mentioned water phase and oil phase are mixed in a volume ratio of 1:1 to 20.
In Article 17,
A method, characterized in that the above polyvalent metal compound salt is at least one divalent to tetravalent metal compound salt selected from the group consisting of Zr ²⁺ , Zr ⁴⁺ , Fe ²⁺ , Fe ³⁺ , Ca ²⁺ , Al ³⁺ , Cu ²⁺ , and Zn ²⁺ .
In Article 17,
A method, characterized in that the above emulsion is added to a polyvalent metal compound salt aqueous solution in a volume ratio of 1:1 to 20.
In Article 17,
The above method is characterized in that the size distribution of microspheres is controlled by controlling the rpm of the homogenizer.
A method for preparing a composition for treating cancer, comprising the following steps:
A step of forming an emulsion by mixing an aqueous phase containing an anionic polymer and an oil phase containing a nonionic surfactant using a homogenizer;
A step of forming microspheres by adding the above emulsion to a multivalent metal compound salt aqueous solution;
A step of separating microspheres of a desired size using a sieve; and
A step of mixing a cationic drug with the microspheres.
Microspheres comprising an anionic polymer; and a polyvalent metal compound salt.