WO2019156477A1 - Composition for hepatic artery embolization, containing microbubbles carrying water-soluble anti-cancer drug, and preparation method therefor - Google Patents

Abstract

Provided in the present invention are a composition for hepatic artery embolization, containing an embolic material and microbubbles carrying a water-soluble anti-cancer drug, a method for applying the composition to hepatic artery embolization, and a preparation method therefor.

Description

A composition for hepatic artery embolization comprising a microbubble carrying a water-soluble anticancer agent and a method of manufacturing the same

The present invention relates to hepatic artery chemoembolization and to increase the effect of hepatic artery embolization using a microbubble carrying a water-soluble anticancer agent.

This application claims priority based on Korean Patent Application No. 10-2018-0015930 filed on February 8, 2018, and all the contents disclosed in the specification and drawings of the application are incorporated in this application.

Recent advances in imaging technology have made it possible to find the exact location of cancer hiding in the body and to remove it in various ways, including irradiation and endoscopy. However, even if the location of the cancer is correctly detected, surgical removal may not be possible due to various reasons, such as when the cancer is spread to all organs or attached to other organs. Even if liver cancer or pancreatic cancer is found, surgical repair is often impossible.

Currently, chemoembolization, the most frequently used procedure for the treatment of liver tumors, is to find arteries that supply liver tumors, administer anticancer drugs, and block the vessels. Liver tissue is supplied with oxygen and nutrients through the portal vein and small hepatic arteries directly from the small intestine and the large intestine. Normal liver tissue is mainly from the portal vein and tumor tissue is mainly from the hepatic artery. Therefore, administering an anticancer agent to the hepatic artery that nourishes the tumor and blocking the blood vessel can selectively necrosis the tumor without harming normal liver tissue. These treatments have a wide range of application because there is no restriction according to the progression of cancer, and there are many advantages such as the limitation of treatment target is the most contributing method to improving liver cancer treatment rate. Chemoembolization first involves catheter insertion into the femoral artery located in the inguinal area, accesses the hepatic artery, and is then injected with angiography to obtain the necessary information such as the location, size and blood supply of the tumor. Insert a thin tube into the catheter to find the target artery.

However, according to the conventional hepatic artery chemoembolization, there is a problem that the embolic material is washed off after a certain time, there is a problem that circulates along the blood vessels and spreads the drug throughout the body, accompanied by side effects such as the death of normal cells.

In order to solve this problem, the present inventors have confirmed that the effect of hepatic artery embolization is increased by using a microbubble that effectively supports it instead of the water-soluble anticancer agent currently used for hepatic artery embolization.

Since the embolic material used for conventional hepatic artery embolization is washed out after a certain period of time after embolization, the present invention solves this problem and at the same time sustains the long-term effect by continuous drug release, thereby improving the effect of chemotherapy. It is an invention to make.

In addition, the prior art is due to the limitation as described above, there is a problem that the side effects such as death of normal cells to spread the drug throughout the system by circulating along the blood vessel bar to solve this problem.

Specifically, the present invention provides a new composition and method for preparing the same, which are applied to hepatic artery embolization, using a microbubble that effectively supports a water-soluble anticancer agent used in hepatic artery embolization.

One embodiment of the present invention is an embolic material; It provides a composition for liver artery embolization and a method for applying the composition for hepatic artery embolization comprising a microbubble carrying a water-soluble anticancer agent.

The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent may be 1: 1 to 1: 4.

Another embodiment of the present invention is an embolic material; Water-soluble anticancer agents; It provides a composition for liver artery embolization and microbubble; and a method for applying the composition for hepatic artery embolization to hepatic artery embolization.

The volume ratio of the water-soluble anticancer agent to the embolic material may be 1: 1 to 1: 4.

The embolic material may be at least one selected from the group consisting of Lipiodol, Absolute ethanol, NBCA (N-butyl-2-cyanoacrylate, Sodium tetradecyl sulfate (sotradecol)).

The water-soluble anticancer agent may be one or more selected from the group consisting of mitomycin, cisplatin, adriamycin, and gemcitabine.

The concentration of the water-soluble anticancer agent may be 1-100 mg / mL.

Another embodiment of the invention provides a microbubble loaded with a water-soluble anticancer agent; It provides a method for producing a composition for hepatic artery embolization comprising; and mixing the microbubble and the embolic material.

The microbubble loaded with the water-soluble anticancer agent may be prepared by dispersing the water-soluble anticancer agent in Computed Tomography (CT) and X-ray contrast agent; And adding and dispersing the dispersed water soluble anticancer agent to the lipid film.

The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent may be 1: 1 to 1: 4.

In addition, another embodiment of the present invention comprises the steps of dispersing a water-soluble anticancer agent in Computed Tomography (CT) and X-ray contrast agent; Mixing the dispersed water-soluble anticancer agent with an embolic material; And mixing the formed mixture with the microbubble; provides a method for preparing a composition for hepatic artery embolization.

The volume ratio of the dispersed water-soluble anticancer agent to the embolic material may be 1: 1 to 1: 4.

The Computed Tomography (CT) and X-ray contrast agent is Iopamidol, Iobitridol, Iopromide, Iodixanol, Iohexol, Ioexol, Ioversol) and Iomeprol may be one or more selected from the group consisting of.

The embolic material may be at least one selected from the group consisting of Lipiodol, Absolute ethanol, NBCA (N-butyl-2-cyanoacrylate and Sodium tetradecyl sulfate (sotradecol)).

In addition, the water-soluble anticancer agent may be one or more selected from the group consisting of mitomycin, cisplatin, adriamycin, and gemcitabine.

The composition for hepatic artery embolization of the present invention not only effectively releases anticancer agents from target cells by using microbubbles carrying water-soluble anticancer agents, but also provides excellent contrast effect by microbubbles.

1 is an embodiment of the present invention, an image of a microbubble carrying a water-soluble anticancer agent.

2 is a schematic view and image of a composition for hepatic artery embolization including a microbubble carrying an embolic material and a water-soluble anticancer agent as an embodiment of the present invention.

3 is a schematic view and image of a composition for hepatic artery embolization including an embolic material, a water-soluble anticancer agent, and a microbubble as an embodiment of the present invention.

4 is an enlarged image of a composition for hepatic artery embolization including an embolic material, a water-soluble anticancer agent, and a microbubble as an embodiment of the present invention.

5 is a graph showing the size distribution of each formulation of the present invention.

First, the concept necessary for practicing the present invention will be briefly described.

Types of distilled water are divided into primary, secondary and tertiary distilled water depending on how many times purified distilled water is applied. Distilled water used in the present invention is tertiary distilled water, which is organic water and ions removed. Tertiary distilled water was used because it is a biologically related invention with a large impact on organic matter.

The liver is an organ that receives a double blood flow from the hepatic artery and portal vein. Normal liver tissue receives 70-80% of the blood flow and 50% of the required oxygen from the portal vein, whereas hepatocellular carcinoma is usually a hypervascular tumor with more than 90% of the blood coming from the hepatic artery. Therefore, injecting a therapeutic substance through the hepatic artery is administered to a hepatocellular carcinoma at a higher concentration than normal liver tissue, and even when the embolus is injected to block the hepatic arterial blood flow non-selectively, severe ischemia is caused only in hepatocellular carcinoma. Is possible. This is the theoretical basis commonly applied to the treatment of hepatocellular carcinoma through the hepatic artery.

Looking specifically to the present invention, in order to solve the above problems, the present invention provides a composition for hepatic artery embolization comprising an embolic material, and a microbubble carrying a water-soluble anticancer agent.

In the composition according to the present invention, the microbubbles carrying the water-soluble anticancer agent may be dispersed in an embolic material. In addition, the microbubbles carrying the embolic material and the water-soluble anticancer agent may form an emulsion.

According to the present invention, the anticancer agent carrying the microbubble carrying the water-soluble anticancer agent can be intensively released to the target cells, and at the same time, the contrast effect by the microbubble can also be provided.

In addition to the water-soluble anticancer agent, the microbubble according to the present invention may additionally carry a contrast agent, for example, an X-ray contrast agent, an MRI contrast agent, a CT contrast agent, and the like. For example, Iopamidol (Pamilay) ^TM ), Iobitridol, Iopromide, Iodixanol, Iohexol, Ioversol, Iomeprol, etc. It is not limited.

The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent may be 1: 1 to 1: 4.

Another embodiment of the present invention provides a composition for hepatic artery embolization comprising an embolic material, a water-soluble anticancer agent and a microbubble, and a method for applying the composition for hepatic artery embolization to hepatic artery embolization.

In the composition according to the present invention, the water-soluble anticancer agent and the microbubble may be dispersed in an embolic material. In addition, the embolic material, the water-soluble anticancer agent and the microbubble may form an emulsion.

The volume ratio of the water-soluble anticancer agent to the embolic material may be 1: 1 to 1: 4.

The embolic material may be one or more selected from, for example, Lipiodol, Absolute ethanol, NBCA (N-butyl-2-cyanoacrylate, Sodium tetradecyl sulfate (sotradecol)), or the embolic material may be useful. It may be an embolic material, for example, may be oil iodide, preferably Lipiodol, but is not limited thereto.

The water soluble anticancer agent may be, for example, one or more selected from the group consisting of mitomycin, cisplatin, adriamycin, and gemcitabine.

The concentration of the water-soluble anticancer agent may be 1 to 100 mg / mL, preferably 1 to 20 mg / mL, more preferably 1 to 10 mg / mL. At this time, the concentration of the water-soluble anticancer agent is a concentration of the water-soluble anticancer agent itself, not the concentration of the composition. The concentration of the water-soluble anticancer agent compared to the composition for hepatic artery embolization may be 1-20mg / mL.

In the composition according to the present invention, various components may be added as long as the object of the present invention is not impaired. For example, the composition according to the present invention may include albumin. The albumin is preferably a water-soluble albumin nanoparticles, it may be included dispersed in the embolic material in the composition. In addition, when albumin is included, the albumin may be included in combination with the microbubble surface according to the present invention, or may be included in a mixture with the microbubble, but is not limited thereto.

Another embodiment of the invention provides a microbubble loaded with a water-soluble anticancer agent; And it provides a method for producing a composition for hepatic artery embolization comprising the step of mixing the microbubble and embolic material.

The microbubble loaded with the water-soluble anticancer agent may be prepared by dispersing the water-soluble anticancer agent in Computed Tomography (CT) and X-ray contrast agent; And adding and dispersing the dispersed water soluble anticancer agent to the lipid film.

The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent may be 1: 1 to 1: 4.

In addition, another embodiment of the present invention comprises the steps of dispersing a water-soluble anticancer agent in Computed Tomography (CT) and X-ray contrast agent; Mixing the dispersed water-soluble anticancer agent with an embolic material; And mixing the formed mixture with the microbubble; provides a method for preparing a composition for hepatic artery embolization.

The volume ratio of the dispersed water-soluble anticancer agent to the embolic material may be 1: 1 to 1: 4.

The Computed Tomography (CT) contrast agent and X-ray contrast agent are Iopamidol (Familay ^TM ), Iobitridol, Iopromide, Iodixanol, Iodixanol (Iodixanol) Iohexol), Ioversol (Ioversol), Iomeprol (Iomeprol) may be one or more selected from the group consisting of, it is variously selected depending on the specific gravity. The selection of specific gravity selects the contrast agent, which allows the embolic material and microbubbles to be evenly dispersed into the emulsion.

In addition, in order to minimize dilution in the mixing process, the microbubbles are preferably subjected to concentration through centrifugation.

The centrifugation is preferably centrifuged for 10 minutes at 225g.

Through the following examples will be described the present invention in more detail. These examples are only for illustrating the present invention in more detail, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. will be.

Production Example  One - Microbubble  Produce

Lipids include 1,2-disteraoyl-sn-glycero-3-phosphocholine (DSPC), DSPE-PEG ₂₀₀₀ -NHS (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-n [poly (ethyleneglycol)] _2000- N-hydroxysuccinimide) was adjusted to molar ratio of 9: 1, dissolved in chloroform, and chloroform was completely evaporated using a rotary evaporator to form a lipid thin film.

Subsequently, 0.01 M of phosphate buffered saline (PBS, pH 7.4) was added to the lipid thin film and the lipids were dissolved while maintaining the temperature at 55-60 ° C. After filling the C ₃ F ₈ gas into the container containing the mixed solution, the microbubble was prepared by applying mechanical agitation for 45 seconds.

Production Example  2 - DOX -loaded Microbubble  in Lipiodol  ( DOX -MB- Lipiodol ) Produce

1. Doxorubicin · HCl was dissolved in pamiray, an X-ray contrast agent, to a concentration of 10 mg / ml. The DOX concentration of DOX-MB dispersed in Pamiray was 6.25 mg / ml. Thereafter, 625 μl (DOX 6.25 mg) and 375 μl pamiray were mixed in DOX in pamiray solution to make a total of 1 ml and placed in a glass vial containing dried lipid film. 30 seconds of sonication was performed in the water bath to sufficiently dissolve the lipid and enough C ₃ F ₈ gas was added to the liquid surface. Thereafter, agitating with a vial mixer, a microbubble loaded with Doxorubicin-HCl was prepared. (DOX-MB)

2. Take 100μl from DOX-MB and 200μl from Lipiodol, put them together in a 1.5ml plastic tube, pipette and vortex for 15 ~ 20 seconds to mix them well. (E.g. DOX-MB: lipiodol = 1: 2, v / v)

3. If layer separation seems to occur with the naked eye, continue vortexing.

Production Example  3- DOX  in Lipiodol  with Microbubble  ( cTACE -MB) manufacturing

1. Doxorubicin · HCl was dissolved in pamiray to 6.25mg / ml concentration. The DOX concentration of cTACE dispersed in Pamiray is 6.25 mg / ml. Thereafter, 100 μl and 200 μl of Lipiodol were added to a 1 ml syringe in DOX in pamiray solution. After that, using a device capable of 3 way pumping, the syringe was counted once to move once and pumped a total of 15 to 20 times. (The formulation of cTACE formulation is assumed to be DOX in pamiray: lipiodol = 1: 2, (v / v))

2. Prepare a microbubble (0.5mg / ml or 1mg / ml) in a 5ml syringe and put 1ml of PBS in a glass vial containing the microbubble to recover the remaining microbubble and put in a 5ml syringe.

3. A 5 ml syringe containing microbubbles was placed in a centrifuge (225g, 10 minutes) to collect the microbubbles on the upper layer under the conditions. After that, the microbubble of the top layer was left and the solution in the syringe was discarded.

4. Pipette enough reaction to mix cTACE and microbubbles as needed. After observing with the naked eye was vortexing for 15 to 20 seconds.

5. If layer separation seems to occur with the naked eye, vortexing is continued.

Experiment result

Experimental Example  One - DOX -MB- Lipiodol  Analysis of the Formulation

Figures 1 and 2 show the results of the image analysis for the formulation prepared for the administration of 0.5 mg DOX per rabbit.

FIG. 1 is an image result of a microbubble prepared by dissolving 1 mg of DOX in 1 ml of pamiray and putting 1 ml into a lipid film for forming a microbubble. In the DIC image, the microbubbles were formed properly. When the fluorescence of the DOX wavelength was confirmed, the anticancer drug DOX was loaded properly in the hydrophilic portion of the microbubbles.

Purification was not performed to filter out unloaded DOX to achieve therapeutic concentrations for embolization.

In addition, the portion indicated by the red dotted line in Figure 2 is an emulsion is formed there is a microbubble carrying an anticancer agent therein. (E.g. DOX-MB in Lipiodol)

Here, since DOX-MB is present in the emulsion, it can be seen that microbubbles appear in black inside the emulsion when confirmed by DIC. Could.

Experimental Example  2 - cTACE Analysis of -MB Formulations

3 and 4 show the results of the image analysis of the formulation prepared for the administration of 0.5 mg of DOX per rabbit.

In FIG. 3, the portion indicated by the red dotted line has an emulsion formed therein, and the anticancer agent and the microbubble are present therein. (E.g. cTACE-MB)

In FIG. 3, the anticancer drugs DOX and microbubbles are present in the emulsion, respectively, and when confirmed by DIC, reddox DOX and microbubbles appearing in black are present in the emulsion and observed by fluorescence at the DOX wavelength. It can be observed that the fluorescence due to DOX is strong inside the emulsion.

In addition, FIG. 4 is an enlarged observation of the cTACE-MB formulation by 40 times. The presence of reddish DOX and black microbubbles inside the emulsion can be clearly observed. When observed with the fluorescence of the DOX wavelength, it can be seen that the fluorescence due to DOX appears strongly inside the emulsion.

Experimental Example  3-Size distribution of each formulation

A total of five or more images were obtained for each formulation and then the average size of the formulation was obtained using a scale bar for each magnification on a microscope program.

The above results show the size distribution for the overall schematic (DOX-MB-Lipiodol, cTACE-MB).

As a result of measuring the size of each formulation from the DIC image, it was found that DOX-MB-Lipiodol had an average size of 15.41 ± 9.39 μm and cTACE-MB of 11.87 ± 5.29 μm.

Figure 5 graphically shows the results on the size of each formulation summarized in a table.

The composition for hepatic artery embolization of the present invention is excellent in industrial value because it not only effectively releases anticancer agent from target cells by using microbubbles carrying water-soluble anticancer agents, but also provides excellent contrast effect by microbubbles.

Claims (17)

1. Embolic material; And

   Microbubble carrying a water-soluble anticancer agent; comprising, composition for hepatic artery embolization.
2. Embolic material;

   Water-soluble anticancer agents; And

   Microbubble; comprising, composition for hepatic artery embolization.
3. The method according to claim 1 or 2,

   The embolic material is at least one selected from the group consisting of Lipiodol, Absolute ethanol, NBCA (N-butyl-2-cyanoacrylate, and Sodium tetradecyl sulfate (sotradecol)), composition for hepatic artery embolization.
4. The method according to claim 1 or 2,

   The water-soluble anticancer agent is at least one selected from the group consisting of mitomycin, cisplatin, adriamycin, and gemcitabine, composition for hepatic artery embolization.
5. The method of claim 1,

   The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent is 1: 1 to 1: 4 composition for hepatic artery embolization.
6. The method of claim 2,

   The volume ratio of the water-soluble anticancer agent to the embolic material is 1: 1 to 1: 4, composition for hepatic artery embolization.
7. The method according to claim 1 or 2,

   The concentration of the water-soluble anticancer agent is 1 ~ 100mg / mL, composition for hepatic artery embolization.
8. Providing a microbubble loaded with a water-soluble anticancer agent; And

   Mixing the microbubble and the embolic material; Method of producing a composition for hepatic artery embolization comprising a.
9. The method of claim 8,

   The microbubble loaded with the water-soluble anticancer agent

   Dispersing the water-soluble anticancer agent in Computed Tomography and X-ray contrast agent; And

   Forming, comprising the step of adding the dispersed water-soluble anticancer agent to the lipid film and then melting and mixing, hepatic artery embolization composition.
10. Dispersing the water-soluble anticancer agent in Computed Tomography and X-ray contrast agent;

    Mixing the dispersed water-soluble anticancer agent with an embolic material; And

    Mixing the mixture and the microbubble formed; method for preparing a composition for hepatic artery embolization.
11. The method of claim 9 or 10,

    Computed Tomography and X-ray contrast agents are Iopamidol, Iobitridol, Iopromide, Iodixanol, Iohexol, Ioversol and Iosol Method for producing a composition for hepatic artery embolization, at least one selected from the group consisting of meprol (Iomeprol).
12. The method of claim 8 or 10,

    The embolic material is at least one selected from the group consisting of Lipiodol, Absolute ethanol, NBCA (N-butyl-2-cyanoacrylate and Sodium tetradecyl sulfate (sotradecol)), a method for producing a composition for hepatic artery embolization.
13. The method of claim 8 or 10,

    The water-soluble anticancer agent is at least one selected from the group consisting of mitomycin, cisplatin, adriamycin, and gemcitabine, method of producing a composition for hepatic artery embolization.
14. The method of claim 8,

    The volume ratio of the microbubble to the embolic material carrying the water-soluble anticancer agent is 1: 1 to 1: 4 method for producing a composition for hepatic artery embolization.
15. The method of claim 10,

    The volume ratio of the dispersed water-soluble anticancer agent to the embolic material is 1: 1 to 1: 4, the method of producing a composition for hepatic artery embolization.
16. Embolic material; And a microbubble carrying a water-soluble anticancer agent.
17. Embolic material; Water-soluble anticancer agents; And microbubbles; a method comprising applying to a hepatic artery embolization.

Images