KR20030023470A - Oily paclitaxel composition and formulation for chemoembolization and preparation method thereof - Google Patents

Abstract

PURPOSE: Provided are paclitaxel oil composition made by oily contrast medium for chemical coloring and the preparation method thereof. The composition has an advantage of delivering the anti-cancer agent, paclitaxel, to the target site because it shows color contrast in the vessel, making it applicable in artery coloring for the treatment of the solid type cancer such as liver cancer. CONSTITUTION: The oil composition comprises: 0.0001mg-10mg of paclitaxel in 1 ml of oily contrast medium selected from a group of iodized poppy seed oil, ethiodol and iodized soybean oil, wherein iodine contents are 30-50wt.%; 0.1-1 ml (based on 1 ml of oily contrast medium) of plant or animal oil such as soybean oil or squalene; sterilizing with EO gas or gamma ray; and making one phase product with a viscosity of 40-180cP at room temperature. A long time storage product consists of 0.0001-13ml paclitaxel, 1ml oily contrast medium and 0.01-1ml precipitation inhibitor selected from a group of alcohol, polyol, oil, lipid, polymer and peptide.

Description

Oil-based compositions for chemical coloration paclitaxel solubilization, formulations thereof and preparation method thereof {OILY PACLITAXEL COMPOSITION AND FORMULATION FOR CHEMOEMBOLIZATION AND PREPARATION METHOD THEREOF}

The present invention relates to an oil-based composition for solubilizing paclitaxel, solubilizing paclitaxel and using it for arterial embolization, a formulation thereof, and a method for preparing the same.

Arterial embolization is a method of treating cancer by finding an artery that goes to the cancer site using a contrast agent and blocking nutrient supply to cancer tissues using anticancer agents and embolic materials.

Representative arterial embolization is hepatic artery embolization. This method involves dispersing various anticancer drugs such as doxorubicin (Adriamycin), cisplatin, and carboplatin in an oily contrast medium, and simultaneously administering embolization and contrast. .

Representative contrast agents used in such arterial embolizations are iodized oils such as lipiodol. However, the dispersion system in which the various anticancer agents are dispersed in Lipiodol has many limitations during the procedure because of physical instability.

In addition, the anticancer agents widely used in the liver cancer treatment method which is conventionally treated in the diagnostic radiology department include adriamycin, epirubicin, and the like. Most of the anticancer drugs including adriamycin cannot be directly dissolved in Lipiodol. The procedure was used in the form of a suspension. (Yoshihiro Katagiri et al., Cancer Chemother. Pharmacol 1989, 23, 238-242). However, in the case of the suspension made by this method, long-term storage is difficult because adriamycin particles are aggregated or precipitates are formed. Therefore, a method of dissolving an anticancer agent in an aqueous contrast agent and dispersing it in Lipiodol, an oily contrast agent, has been used. That is, just before administration to the patient, the anticancer agent is dissolved in the aqueous contrast agent and mixed with the oily contrast agent by the pumping method. At this time, in order to maximize the stability of the emulsion, an aqueous contrast agent Eurographine (Urografin, 1.328-1.332) or Iopamiro, which has a specific gravity similar to that of Lipiodol (1.275 to 1.290), is used (Takashi Kanematsu Et al., Journal of surgical oncology 1984, 25, 218-226, Takafumi Ichida et al., Cancer Chemother.Pharmacol 1994, 33, 74-78).

However, by this method, only a temporary emulsion is formed, and the mixed state is unstable, so that the separation occurs again in a few minutes, and thus adriamycin precipitates, so that it is hardly absorbed by the body. Indeed, it can be observed that phase separation occurs during injection into the patient's blood vessels during the procedure. In addition, when the mixed solution is administered, adriamycin is absorbed at once, so antitumor effects cannot be expected continuously, and various side effects appear.

To date, the best known method for liver cancer treatment is a synthetic polymer called poly (styrene-co-maleic acid) -conjugated neocarzinostatin (SMANCS)], a poly (styrene-co-maleic acid) -conjugated neocarzinostatin. SMANCS has both hydrophilic and lipophilic properties and can be directly dissolved in Lipiodol (Konno, T. and Maeda, H., Targetting chemotherapy of hepatocellular carcinoma.Neoplasms of the liver, Eds. Okuda, K., and Ishak, KG, Springger-Verlag, Berlin, P343-352). Although the SMANCS / lipiodol formulation solves the problems of the adriamycin / lipiodol formulation, it is 10 times more expensive than the adriamycin / lipiodol formulation, and the toxicity is also severe.

Meanwhile, paclitaxel, an anticancer agent, is known to exhibit significant cytotoxicity against various cancers such as ovarian cancer, breast cancer, esophageal cancer, melanoma, and leukemia. Taxol, which is mainly marketed by Bristol-Myers Squibb It is used as a brand injection.

Paclitaxel, a representative poorly soluble drug, has undergone simultaneous development of solubilization technology due to its low solubility. An example of such a solubilization technique is the use of a dissolution aid to use paclitaxel for systemic administration routes such as intravenous injection. Taxol mentioned above Injectables also use a dissolution aid called Cremophor EL (polyoxyethylene 35 castor oil). In other words, this formulation is spontaneously formed in microemulsion when dispersed in water as a pre-concentrate form of emulsion (see US Pat. No. 5438072). However, the use of such dissolution aids is known to cause further toxicity. Therefore, there is a need to develop a new formulation of paclitaxel excellent anti-cancer effect against various cancers.

To meet this need, the inventors of the present invention have studied paclitaxel formulations that can be used for arterial embolization as a new formulation of paclitaxel. Unexpectedly, paclitaxel is well dissolved in oily constrast medium, resulting in a single phase viscosity range of 40 at room temperature. It was found to be a highly viscous oil of -180 cP (centipoises).

It has also been found that the paclitaxel / oil preparation composition is chemically and physically stable and can be preserved for a long time without a change in composition. Thus prepared paclitaxel / oily contrast agent composition exhibits significantly improved physical properties compared to the conventional Lipiodol formulations using water-soluble anticancer agents such as doxorubicin, and can be said to have similar formulation properties as the SMANCS / Lipiodol formulation. While conventional SMANCS / lipiodol formulations are expensive and are not universalized due to their severe toxicity, the paclitaxel / oily contrast agent composition of the present invention has a low production cost due to the low cost of the two materials as raw materials and a very high manufacturing process. Simple and obtained formulations also have excellent stability and shelf life.

That is, the composition of the present invention can maintain the original composition stably over several hours of arterial embolization, so that the application of arterial embolization when compared with the conventional Lipiodol / Iopamiro / Adriamycin preparations cause phase separation immediately after mixing When the anticancer agent is stably delivered to the body for a long time, as well as the shelf life of the formulation itself is excellent. In addition, as described below, as a result of performing hepatic artery embolization using an animal model, it is expected to be widely used for arterial embolization because it shows excellent embolism effect and anticancer action.

Although arterial embolization is typical of hepatic artery embolization, it can be applied to various types of solid cancers in addition to hepatic artery embolization. For example, SMNACS / lipiodol formulations can be used to perform cancer therapy that targets only cancer cells, such as the treatment of rectal cell carcinoma through the rectal artery (K. Tsuchiya, Tumor-targeted chemotherapy with SMANCS in lipiodol). for renal cell carcinoma: longer survival with larger size tumors.Urology. 2000 Apr; 55 (4): 495-500).

To date, there has been no use of paclitaxel as an anticancer agent used in vascular embolization or contrast. Accordingly, the present invention intends to dissolve paclitaxel in an oily contrast agent and use it for arterial embolization.

Accordingly, one object of the present invention is to provide a novel paclitaxel oily composition comprising an oily contrast agent, in which paclitaxel is solubilized.

Another object of the present invention is to provide a preparation of oily paclitaxel for use in arterial embolization for the treatment of solid cancer. In other words, to provide a stable oil-based paclitaxel formulation that can maintain the original composition through the arterial embolization procedure for several hours.

Another object of the present invention to provide a method for producing the paclitaxel oil composition described above.

1 is a CT photograph of one week after the paclitaxel / lipiodol formulation 0.3 CC of the present invention is selectively administered to liver cancer tissues of rabbits by hepatic artery embolization. The amount of paclitaxel administered corresponds to A) 1 mg, B) 3 mg and C) 0 mg.

Figure 2 is selectively administered paclitaxel / Lipiodol formulation of the present invention to liver cancer tissues of rabbits by hepatic artery embolization, and after one week, paclitaxel concentrations in liver cancer tissues and normal tissues are quantified and shown by HPLC. The amount of paclitaxel administered corresponds to A) 1 mg and B) 3 mg.

Figure 3 is a paclitaxel / Lipiodol formulation of the present invention 0.3 CC (1 mg group and 3 mg group) E is 0.4 CC (4 mg group) is selectively administered to liver liver tissue of rabbits by hepatic artery embolization 1 week after the total liver cancer tissue Shows the percentage of living cells among them. Only 0.3 cc of Lipiodol was administered to the control group.

Figure 4 is selectively administered paclitaxel / Lipiodol formulation 0.4 CC (paclitaxel 4 mg) of the present invention to the liver cancer tissue of rabbits by hepatic artery embolization and shows the paclitaxel concentration in liver cancer tissue, left lobe, right lobe for 1 week.

-●-; Paclitaxel concentration in liver cancer tissue,

-○-; Concentration of paclitaxel in the left lobe,

-▲-; Paclitaxel concentration in the right lobe.

5 is a photograph after storage of the paclitaxel / lipiodol formulation and the paclitaxel / lipiodol / tricapryline formulation of the present invention at room temperature for 90 days or under a polarizing microscope.

A; Picture of Paclitaxel / Lipiodol Formulation

B; Photos under polarized light microscope of paclitaxel / lipiodol formulation

C; Photos of paclitaxel / lipiodol / tricapryline formulation

D; Photograph under polarized light microscope of paclitaxel / lipiodol / tricapryline formulation

Figure 6 is a specific graph of the thickness of the sole after inoculating melanoma cells 20 days (200 μg of paclitaxel) formulation of the paclitaxel / Lipiodol / Tricapryline formulation of the present invention to the rat's foot after 5 days. As a control group, a group injected with 20 μl of Lipiodol / Tricapryline and a group without any treatment were used.

-●-; 20 μl of paclitaxel / lipiodol / tricapryline formulation (200 μg of paclitaxel),

-○-; 20 μl of Lipiodol / Tricapryline formulations were injected,

-▲-; You haven't done anything.

FIG. 7 shows the population of live mice over time after inoculating melanoma cells and injecting 20 μl of paclitaxel / lipiodol / tricapryline formulation of the present invention (200 μg of paclitaxel) into the paws of mice. One graph. As a control, a group without any treatment was used.

-●-; 20 μl of paclitaxel / lipiodol / tricapryline formulation (200 μg of paclitaxel),

-○-; You haven't done anything.

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a paclitaxel oil-based composition for chemical coloration solubilized with paclitaxel using an oily contrast agent to be used for chemical embolization, and a method for preparing the same. In addition, the present invention relates to a paclitaxel oil-based composition for chemical color, which further contains a substance that inhibits the precipitation of paclitaxel for long-term storage, and a method for preparing the same. Since the composition of the present invention is solubilized paclitaxel, angiography can be performed during chemical embolization, the anticancer agent may be delivered to target cells when the chemical embolization is performed.

An example of an oily contrast agent usable in the paclitaxel oil composition of the present invention is iodide oil. Typical iodide oils include iodide poppy seed oil or iodide soybean oil, such as Lipiodol Laboratoire Guerbet, France, ethiodol (Savage Laboratories, Melville, NY). Can be. For more information on iodide soybean oil, see Ma Tai. The effect of oral iodized oil on prevention and treatment of endemic goiter. Chienese Med. J. 61 (9): 533, 1981.

In the present invention, the iodine content of the iodide oil which can be used as an oily contrast agent is preferably 30 to 50% by weight. More preferably, the iodine content is 35 to 45% by weight. Most preferably, Lipiodol is preferably used as an oily contrast agent.

The paclitaxel oil-based composition for chemical coloration of the present invention contains paclitaxel in an amount of 0.0001 mg to 10 mg per 1 ml of an oily contrast agent and an oily contrast agent. At this time, if the amount of paclitaxel to 1 mg of the oily contrast agent is more than 10 mg, it is not dissolved in the oily contrast agent any more, and precipitation occurs, and if the amount of paclitaxel is 0.0001 mg or less, it is not preferable because the desired anticancer effect cannot be achieved. .

In addition, the paclitaxel oil-based composition of the present invention may optionally additionally contain an animal oil such as squalene or a vegetable oil such as soybean oil. That is, by replacing a part of the oily contrast agent with such animal oil, vegetable oil, or a mixture thereof, it is possible to reduce the manufacturing cost while maintaining biocompatibility without reducing the desired drug efficacy or stability. The amount of the animal oil or vegetable oil is used in a ratio of oily contrast agent: animal oil and / or vegetable oil = 1: 0.01 to 1 by volume. Preferably the ratio is 1: 0.01 to 0.5.

The paclitaxel oily composition of the present invention can be easily prepared by first adding an appropriate amount of paclitaxel to the oily contrast agent so as to be in the above composition range, stirring and dissolving at room temperature. At this time, for rapid dissolution, it may be heated to a temperature of about 35 ~ 45 ℃ or sonication using a bath type sonicator (bath type sonicator). The paclitaxel oily composition thus prepared is stored after sterilization. As the sterilization method, a method of sterilization using a raw material sterilized from the beginning or mixed under sterilization conditions or using a syringe filter (pore size 200 μm, PVDF sterilization filter) may be used. As another sterilization method, iodized poppy seed oi and paclitaxel are sterilized with gamma-ray, sterile via EO gas sterilization or mixed, or after mixing the composition is gamma-ray sterilized or EO gas sterilized It can also be disinfected by the method.

The paclitaxel oil composition of the present invention thus prepared showed stable storage stability at room temperature for at least 60 days.

In the oily composition, paclitaxel is well dissolved in an oily contrast agent, and thus is stably stored for about 2 months, but paclitaxel precipitates and precipitates over time. This phenomenon is due to the formation of intramolecular or intermolecular hydrogen bonds between hydroxy groups in the paclitaxel molecule. This phenomenon can be prevented by adding a small amount of a substance that forms hydrogen bonds with paclitaxel or a substance that prevents intermolecular hydrogen bond formation of paclitaxel. Therefore, if the oily contrast agent used in the oily composition can form hydrogen bonds with paclitaxel, precipitation of paclitaxel does not occur even after two months.

However, Lipiodol among the oily contrast agents usable in the present invention cannot form hydrogen bonds due to its molecular structure, and thus, hydrogen bonds between paclitaxel molecules cannot be prevented with time. In such a case, the addition of a small amount of a substance that forms a hydrogen bond well while soluble in Lipiodol, such as tricapryline, forms a hydrogen bond between paclitaxel-tricapryline instead of the hydrogen bond between paclitaxel and paclitaxel. You can stop it.

In the case of the oily composition of paclitaxel of the present invention, the content of long-term storage of paclitaxel and the oily contrast agent is determined by the manufacturing process. If there is no contact with moisture or oxygen during the manufacturing process and need not be heated, the components can be prevented from being oxidized, thus allowing for longer storage. However, the formation of paclitaxel precipitation from the paclitaxel oily composition of the present invention is a change in thermodynamically stable form, and thus a problem that occurs during long-term storage regardless of whether or not care is taken in preparation. The precipitation formation rate was different depending on the concentration of paclitaxel in the paclitaxel oil composition, but it was found that precipitation was formed at room temperature over 60 days for 10 mg / ml and 120 days for 5 mg / ml. Therefore, in order to make the shelf life more than 1 year practical, it is only possible to maintain a thermodynamically stable state for a long time only by adding a substance that inhibits the formation of paclitaxel.

Therefore, the paclitaxel oily composition of the present invention may further contain a substance that inhibits the precipitation formation of paclitaxel. At this time, the solubility of paclitaxel in the oily contrast agent is increased to 13 mg / ml.

That is, the paclitaxel oil composition of the present invention may contain a substance for inhibiting precipitation formation of 0.0001 to 13 mg of paclitaxel and 0.01 to 1 ml of paclitaxel relative to 1 ml of the oily contrast agent and the oily contrast agent.

The oily contrast agent used at this time is as described above.

As a material for inhibiting the formation of paclitaxel, a substance capable of hydrogen bonding with the paclitaxel molecule or a substance that prevents hydrogen bond formation between the paclitaxel molecules may be used.

Substances capable of hydrogen bonding with the paclitaxel molecules include alcohols, polyols, oils, lipids, polymers or peptides. The alcohol includes methanol, ethanol, propanol, isopropanol, butanol and the like. Polyols include ethylene glycol, propylene glycol, and polyethylene glycol. Such oils include naturally occurring animal or vegetable oils that are triglycerides, diglycerides, monoglycerides, tocopherols or mixtures thereof. The lipids include phospholipids, neutral lipids, cationic lipids or anionic lipids. The polymer is poly lactide-co-glycolic acid (PLGA).

Dimethylsulfoxide (DMSO) or an amide may be used as a chaotropic agent to prevent the formation of hydrogen bonds between the paclitaxel molecules.

Thus, paclitaxel oil-based composition containing a substance that inhibits the formation of paclitaxel showed stable storage stability at room temperature for more than 200 days.

The paclitaxel oily composition of the present invention is useful as a composition for chemoembolization for the treatment of solid cancer, and has a viscosity suitable for chemoembolization of 45 to 180 cP. The dosage and administration method of the paclitaxel oil-based composition of the present invention used for chemoembolization may be varied at the discretion of the attending physician according to the individual's individual differences such as the patient's age, sex, weight, and severity of symptoms. The procedure can be repeated several times, once every four months, and infusions of 2-15 ml at a time are injected into the artery that supplies solid cancer, ie liver cancer, along the hepatic artery.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the examples set forth below are provided to aid the understanding of the present invention, but the present invention is not limited to these examples.

Example 1.

Preparation of Paclitaxel / Lipiodol Oil Composition

1 ml of Lipiodol (Lipiodol Ultra-fluid, Laboratoire Guerbet, France, 38% by weight of iodine) was added to 2,4,6,8,10,11 mg of paclitaxel (Samyang Corp.). Into a micro test tube, 1.5 ml polyethylene, Eppendorf AG, Germany) and dissolved by stirring at room temperature. For rapid dissolution, it may be heated to about 40 ° C. or sonicated in a bath sonicator. When the concentration of paclitaxel was 2 to 10 mg / 1 ml Lipiodol, paclitaxel was dissolved in Lipiodol because a single liquid phase was formed. However, if the paclitaxel concentration is 11 mg / 1 ml Lipiodol, paclitaxel initially dissolves in Lipiodol to form a single liquid phase, but if left for one day, the turbidity of the solution increases, and the paclitaxel in the Lipiodol is observed due to the formation of a precipitate of paclitaxel under a microscope. It was confirmed that the solubility of about 10 mg / ml at room temperature (24 ~ 28 ℃). Viscosity was measured by adding 10 ml of paclitaxel / lipiodol obtained in a kinematic viscometer (Cannon-Fenske Type, Calibrated, Cat.No. 13-617E, Size 200, Fisher Scientific, Pittsburgh, PA) and measuring the drop time. . The measured viscosity is 67 cP, and the paclitaxel / lipiodol composition is expected to have a high embolic effect when the viscosity is 45 cP or more.

Example 2.

Physical Stability of Paclitaxel / Lipiodol Composition

As an oily contrast agent, 10 mg of paclitaxel is added to 1 ml of Lipiodol (38% by weight of Lipoiodol Ultra-fluid, Laboratoire Guerbet, France, Iodine), and the solution is stirred at room temperature and dissolved. Warmed to about 40 ° C. to accelerate dissolution. It can be seen that paclitaxel is dissolved in Lipiodol because a single liquid phase is formed after preparation. The prepared composition was sterilized by a syringe connected to a syringe filter (200 μm pore size, PVDF filter), and then stored at room temperature and 4 ° C. for 60 days to observe physical stability and chemical degradation of paclitaxel. Color change, odor, and phase separation of the liquid phase did not occur. Analysis of the components using HPLC showed that paclitaxel did not degrade at all. HPLC conditions were as follows.

SP8810 precision isocratic pump (Spectra-Physics Inc., San Jose, CA)

Column: Waters Bondpack C18 Column (3.9 mm x 300 mm, Waters Corp., Milford, MA)

Mobile phase: 50% (w / w) each of acetonitrile and water

Flow rate: 1 ml / min

Detector: Spectra 100 variable wavelength detector (Spectra-Physics)

Example 3.

Preparation and Physical Stability of Ethiodol / Paclitaxel Compositions

Paclitaxel oil-based composition was prepared in the same manner as in Example 1 except that thiodol (Savage Laboratories, Melville, NY) was used instead of Lipiodol as an oily contrast agent. It can be seen that paclitaxel is well soluble in thiodol because the mixture forms a single liquid phase.

Physical stability of the prepared composition was tested in the same manner as in Example 2. After sterilization, stored at room temperature and 4 ° C. for 60 days, and physical stability and chemical degradation of paclitaxel were observed. Color change, odor, and phase separation of the liquid phase did not occur. Analysis of the components using HPLC showed that paclitaxel did not degrade at all.

Experimental Example 1.

Production of Liver Cancer Animal Model

10ml of rabbit (VX-2 animal model of rabbit liver carcinoma) with a tumor of 1-2cm diameter after about 2 weeks after transplantation of VX2 carcinoma from Deutsches Krebsforschungszentrum Tumorbank, Germany into the femur of rabbit (Newzealand White) Was injected intravenously with pentotal sodium solution (62.5mg / kg) to remove the hair around the tumor, disinfect the skin with iodine solution and alcohol, and cut the skin around the tumor with a surgical knife It peeled off with tissue and extracted. Excision of the tumor was removed to remove necrosis in the center, and a thin layer around the tumor was collected and mixed with calcium and magnesium-free Hank's balanced salt solution (Grand Island Biological Co., Grand Island, New York) to kill tumor tissue. Cut into fine pieces with scissors and surgical knives until 5ml RMPI-1640 (Rosewell Park Memorial Institute, Rosewell Park, New York) culture medium and mixed well and diluted to a concentration of 1x10 ⁶ / ml tumor cells.

Intrahepatic Injection and Transplantation of Tumor Cell Solution: To anesthetize rabbits, intravenous injection of 23G needle into the vein of the ear to connect with 500ml saline solution to secure the vein, and then dilute 500mg of pentotal sodium in 40ml saline solution. (1.5 ml / kg) was injected into the ear vein at a rate of 1 ml per minute. The abdominal hair is then removed, sterilized with iodine solution and alcohol, punctured the left lobe of liver using 22G needle under ultrasound guidance, and 0.1ml of tumor tissue solution is injected into parenchyma of liver with 1ml syringe with 22G needle. It was. Tumor tissue was injected into the front lobe of the five lobes of the liver, where ultrasound was most easily observed (FIG. 2). Antibiotics (PenbrexR, 250 mg) were injected intravenously to prevent secondary infection. Rabbits that had been injected with tumor tissue solutions were raised in normal feed in rabbits. Two weeks after tumor cell transplantation, tumors were detected by ultrasound and CT, and the growth curve was used to predict the future growth rate to some extent. After tumor cell transplantation, ultrasound was performed at 3 days intervals from 2 weeks and CT at 1 week intervals to track the location and size of tumors.

Example 4.

Carotid Embolization in Liver Cancer Animal Model Using Paclitaxel / Lipiodol Composition

As an oily contrast agent, 10 mg or 3.33 mg of paclitaxel was added to 1 ml of Lipiodol, respectively, and placed in a test tube, followed by dissolution at room temperature. In order to accelerate the dissolution, it may be heated to about 40 ° C. or sonicated in a bath sonicator. It was found that paclitaxel was dissolved in Lipiodol because a uniform single liquid phase was formed after preparation. The prepared Lipiodol / Paclitaxel formulation was sterilized by passing a syringe connected to a syringe filter (200 μm pore size, PVDF filter). Carotid artery embolization was performed by selecting the artery supplying the tumor to the liver cancer animal model prepared in Experiment 1, using 0.3 ml of the paclitaxel / lipiodol formulation of the present invention using a microconduit. Thus, paclitaxel doses in each experimental group corresponded to 1 mg or 3 mg, respectively. At this time, only 0.3 cc of Lipiodol was injected into the liver cancer animal model as a control group. One week after the surgery, CT scans showed that Lipiodol was selectively observed only in liver cancer tissues as shown in FIG. 1.

Example 5.

Analysis of Paclitaxel Levels in Liver Cancer Tissues after Carotid Embolization with Paclitaxel / Lipiodol Composition

Liver cancer animals embolized in Example 4 were sacrificed one week after the liver was removed. The liver tissues were divided into tissues identified with the naked eye of lipidodol, tissues not identified with the naked eye, and normal tissues located around liver cancer. The separated liver tissue was added to lysis buffer [62.5 mM Tris-HCl (pH 6.8), 2% sodium dodecyl sulfate, 5% β-mercaptoethanol, 10% glycerol] and then ground to destroy the tissue. This was centrifuged to determine the concentration of paclitaxel in the supernatant using HPLC. At this time, the conditions of HPLC are the same as in Example 2. As described in Example 4, quantitative results of paclitaxel in rabbits administered with 1 mg and 3 mg of paclitaxel are shown in FIGS. 2A and 2B, respectively. It was confirmed that the highest concentration of paclitaxel was found in liver cancer tissues in which Lipiodol was visible to the naked eye, and relatively high concentrations of paclitaxel were detected in liver cancer tissues in which Lipiodol was not visible. On the other hand, paclitaxel was selectively distributed only in liver cancer tissues even after one week of administration because paclitaxel concentration was hardly detected in normal liver tissues near liver cancer tissues.

Example 6.

Necrosis rate of liver cancer tissue after carotid embolization with Lipiodol / Paclitaxel composition

As an oily contrast agent, 10 mg or 3.33 mg of paclitaxel was added to 1 ml of Lipiodol, respectively, and placed in a test tube, followed by dissolution at room temperature. Sonication was performed in a bath sonicator to speed up dissolution. It was found that paclitaxel was dissolved in Lipiodol because a uniform single liquid phase was formed after preparation. The prepared Lipiodol / Paclitaxel formulation was sterilized by passing a syringe connected to a syringe filter (200 μm pore size, PVDF filter). The liver cancer animal model prepared in Experiment 1 was supplied with tumors using microconduit with 0.3 ml (10 or 3.33 mg / ml formulation) or 0.4 ml (10 mg / ml concentration formulation) of the paclitaxel / lipiodol formulation of the present invention. Carotid artery embolization was performed. Thus, paclitaxel doses in each experimental group corresponded to 1 mg, 3 mg or 4 mg, respectively. At this time, only 0.3 cc of Lipiodol was injected into the liver cancer animal model as a control group. One week after the surgery, CT scans showed that Lipiodol was selectively detected only in the liver cancer tissues as shown in FIG. 1. Liver cancer animals that underwent embolization were sacrificed one week after the liver was removed. The size of the isolated liver cancer was about the same as that of the control group, which was 32 ± 5 mm in diameter, in the experimental group administered with the paclitaxel / lipiodol formulation. Pathological examination was performed to distinguish between necrotictumor and viable tumor among liver cancer tissues. The percentage of live tumors in all liver cancer tissues is shown in FIG. 3. More than 30% live tumors were identified in the control group treated with Lipiodol alone, but in the experimental group administered with paclitaxel / lipiodol, 13.2%, 10.4%, and 0.6% of paclitaxel doses of 1 mg, 3 mg, and 4 mg, respectively, were observed. It can be seen that the reduction is about 20% or more. These results show that paclitaxel in the paclitaxel / lipiodol formulation effectively kills cancer cells.

Example 7.

Preparation of Lipiodol / Soybean Oil / Paclitaxel Compositions

10 mg of paclitaxel was added to 1 ml of Lipiodol and 0.2 ml of soybean oil, and the mixture was added to a test tube and stirred at room temperature to dissolve. Sonication was performed in a bath sonicator to speed up dissolution. Since the mixture forms a single liquid phase, it can be seen that paclitaxel is dissolved in Lipiodol / soybean oil, which is a mixed oil.

Example 8.

Preparation of Lipiodol / Squalene / Paclitaxel Compositions

Lipiodol / squalene / paclitaxel compositions were prepared in the same manner as in Example 6 except that squalane (squlane) was used instead of soybean oil and warmed at 40 ° C. to accelerate dissolution. Since the mixture forms a single liquid phase, it can be seen that paclitaxel is dissolved in Lipiodol / Squalene, the mixed oil.

Example 9.

Preparation and Physical Stability of Paclitaxel / Lipiodol / Tricapryline Compositions

10 mg of paclitaxel was added to 1 ml of Lipiodol and 0.01 ml of Tricapryline, and placed in a test tube, and stirred at room temperature to dissolve. Sonication was performed in a bath sonicator to speed up dissolution. Since the mixture forms a single liquid phase, it can be seen that paclitaxel is dissolved in Lipiodol / tricapryline, which is a mixed oil. The prepared composition was sterilized by a syringe connected to a syringe filter (200 μm pore size, PVDF filter), and then stored at room temperature and 4 ° C. for 200 days to observe physical stability and chemical degradation of paclitaxel. Color change, odor, and phase separation of the liquid phase did not occur. As a result of analyzing the components using HPLC in the same manner as in Example 2, it was found that paclitaxel was not decomposed at all. At this time, the result of using the paclitaxel / lipiodol composition prepared in Example 1 as a control resulted in turbidity of the oily composition due to the precipitation of paclitaxel at room temperature (FIG. 5A), and precipitation of paclitaxel was observed even under a polarizing microscope. However, in the case of the composition of Example 9 (Fig. 5B), the oily composition remains transparent (Fig. 5C), and no precipitation is observed even under a polarizing microscope, thereby adding tricapryline, a substance that inhibits the formation of paclitaxel. It can be seen that the composition can be stored for a long time.

Example 10.

Preparation and Physical Stability of Paclitaxel / Lipiodol / Tricapryline Compositions

12 mg of paclitaxel was added to 1 ml of Lipiodol and 0.01 ml of Tricapryline, and placed in a test tube, and dissolved by stirring at room temperature. Sonication was performed in a bath sonicator to speed up dissolution. Since the mixture forms a single liquid phase, the addition of tricapryline showed that the solubility of paclitaxel was higher in the Lipiodol / Tricapryline mixed solvent than in Lipiodol.

Experimental Example 2.

Creation of melanoma animal models

B16-F10 cells, a melanoma cell line that spontaneously developed in C57BL / 6J mice, were obtained from a non-US cell line bank, followed by 10% fetal bovine serum (FBS, Gibco BRL / Life Technologies, New York, NY) and 1% Penicillin / Streptomycin. (Gibco) was added to Dulbeccos Modified Eagle Medium (DMEM, Gibco). Melanoma animal models were prepared by inoculating 1 x 10 ⁶ cells in 100 μl of DMEM and inoculating the left foot of 8-week-old C57BL / J mice (Shimtaco, Korea).

Example 11.

Size change rate of melanoma after injection of paclitaxel / lipiodol / tricapryline composition

20 μl of sterile paclitaxel / lipiodol / tricapryline (200 μg of paclitaxel) after passing through a syringe connected to a syringe filter (200 μm pore size, PVDF filter) prepared in Example 9 Five days after the inoculation, the rats were injected into the left plantar inoculation site. At this time, 20 μl of Lipiodol / Tricapryline (100: 1 volume ratio) was injected into the melanoma animal model as a control group and no treatment group was used. The size change of melanoma is measured and the result is shown in FIG. In the untreated group, melanoma began to increase after 18 days of inoculation, and started to increase after only 22 days of the group treated with Lipiodol / Tricaprilin, whereas Paclitaxel / Lipiodol / Tricaprilin was treated. In the group melanoma does not grow at all shows that the composition of the present invention has an anticancer effect.

Example 12.

Size change rate of melanoma after injection of paclitaxel / lipiodol / tricapryline composition

20 μl of sterile paclitaxel / lipiodol / tricapryline (200 μg of paclitaxel) after passing through a syringe connected to a syringe filter (200 μm pore size, PVDF filter) prepared in Example 9 Five days after the inoculation, the rats were injected into the left plantar inoculation site. At this time, the comparison was made using the group not treated with any control. The live population is measured and the results are shown in FIG. In the untreated group, mice began to die about 20 days after inoculation and all individuals died at 48 days (n = 6), whereas in the group treated with paclitaxel / lipiodol / tricapryline, all individuals It shows that the composition of the present invention has an anticancer effect because it survives healthy.

The paclitaxel oil composition of the present invention is a uniform monolithic viscous liquid, and proposes a new dosage form in which paclitaxel, which has been used only as an intravenous injection, is extended to be used for arterial embolization of solid cancer. The paclitaxel / oil-based contrast agent composition is easy to manufacture and sterilize, and its components and composition are physicochemically stable than conventional adriamycin / lipiodol formulations. Therefore, when used for arterial embolization for solid cancer, the stable composition is maintained at room temperature. Stability for at least 60 days. In addition, it is possible to increase the solubility of paclitaxel and maintain stability for at least 200 days by additionally adding a substance that inhibits the formation of paclitaxel.

Claims (35)

Paclitaxel oil composition for chemical color containing 0.0001 mg to 10 mg of paclitaxel per 1 ml of oily contrast agent and oily contrast agent. The paclitaxel oily composition according to claim 1, wherein the oily contrast agent is an iodide oil selected from the group consisting of iodized poppy seed oil, ethiodol, and iodized soylbean oil. The paclitaxel oil composition according to claim 2, wherein the iodine content of the iodide oil is 30 to 50% by weight. The paclitaxel oil composition according to claim 3, wherein the iodine content of the iodide oil is 35 to 48% by weight. The chemical color composition according to claim 1, wherein the oily contrast agent is iodide poppy seed oil having an iodine content of 35 to 48% by weight. The paclitaxel oil composition according to claim 1, which additionally contains animal oil, vegetable oil or a mixture thereof in a ratio of 0.1 to 1 ml with respect to 1 ml of the oily contrast agent. 7. The oily composition of paclitaxel according to claim 6, wherein said animal oil is squalene. 7. The paclitaxel oil composition according to claim 6, wherein the vegetable oil is soybean oil. The paclitaxel oil composition of any one of claims 1 to 8, wherein the viscosity at room temperature is in the range of 40 to 180 cP. 10. The paclitaxel oil composition according to claim 9, which is used exclusively for chemical coloration for treating solid cancer. 11. The paclitaxel oily composition according to claim 10, wherein the solid cancer is liver cancer and is used for hepatic artery embolization. A method for producing a paclitaxel oil composition comprising adding paclitaxel to an oily contrast agent in an amount of 0.0001 mg to 10 mg based on 1 ml of the oily contrast agent, followed by stirring and dissolving at room temperature. 13. The method of claim 12, wherein the sterile oily contrast agent and paclitaxel are dissolved under sterile conditions. 13. The method of claim 12, further comprising the step of dissolving the oily contrast agent and paclitaxel followed by EO gas disinfection or gamma-ray disinfection. 13. The method according to claim 12, wherein the oily contrast agent and paclitaxel are dissolved by warming to a temperature of 35 to 45 캜. 13. The method of claim 12 wherein the sonication dissolves the oily contrast agent and paclitaxel. A long-term storage paclitaxel oil composition containing 0.0001 to 13 mg of paclitaxel and 0.01 ml to 1 ml of paclitaxel with respect to oily contrast agent and 1 ml of oily contrast agent. 18. The paclitaxel oil composition according to claim 17, wherein the oily contrast agent is an iodide oil selected from the group consisting of iodized poppy seed oil, ethiodol, and iodized soylbean oil. 19. The paclitaxel oil composition according to claim 18, wherein the iodine content of the iodide oil is 30 to 50% by weight. 20. The paclitaxel oil composition according to claim 19, wherein the iodine content of the iodide oil is 35 to 48 wt%. 18. The chemical color composition according to claim 17, wherein the oily contrast agent is iodide poppy seed oil having an iodine content of 35 to 48% by weight. 18. The paclitaxel oil composition according to Claim 17, which additionally contains animal oil, vegetable oil or a mixture thereof in a ratio of 0.1 to 1 ml with respect to 1 ml of the oily contrast agent. 23. The oily composition of paclitaxel according to claim 22, wherein said animal oil is squalene. 23. The paclitaxel oil composition according to claim 22, wherein the vegetable oil is soybean oil. 25. The single phase paclitaxel oil composition according to any one of claims 17 to 24, wherein the viscosity at room temperature is in the range of 40 to 180 cP. 27. The paclitaxel oily composition according to claim 25, which is used exclusively for chemical coloration for the treatment of solid cancer. 27. The paclitaxel oily composition according to claim 26, wherein the solid cancer is liver cancer and is used for hepatic artery embolization. 18. The paclitaxel oil composition according to claim 17, wherein the substance which inhibits the precipitation formation of paclitaxel is a substance capable of hydrogen bonding with paclitaxel molecules or a substance which prevents formation of hydrogen bonds between paclitaxel molecules. 29. The paclitaxel oil composition according to claim 28, wherein the substance capable of hydrogen bonding with the paclitaxel molecule is selected from the group consisting of alcohols, polyols, oils, lipids, polymers and peptides. 30. The paclitaxel oil composition according to claim 29, wherein the alcohol is selected from the group consisting of methanol, ethanol, propanol, isopropanol and butanol. 30. The paclitaxel oil composition according to claim 29, wherein said polyol is selected from the group consisting of ethylene glycol, propylene glycol and polyethylene glycol. 30. The paclitaxel oil composition according to claim 29, wherein said oil is selected from the group consisting of naturally extracted animal or vegetable oils which are triglycerides, diglycerides, monoglycerides, tocopherols and mixtures thereof. 30. The paclitaxel oil composition according to claim 29, wherein said lipid is selected from the group consisting of phospholipids, neutral lipids, cationic lipids and anionic lipids. 30. The paclitaxel oil composition according to claim 29, wherein the polymer is poly lactide-co-glycolic acid (PLGA). 29. The paclitaxel oil composition according to claim 28, wherein the chaotropic agent which interferes with the formation of hydrogen bonds between paclitaxel molecules is dimethylsulfoxide (DMSO) or amide.