WO2000076525A1

WO2000076525A1 - Anticancer emulsions containing anthracycline compounds

Info

Publication number: WO2000076525A1
Application number: PCT/JP2000/003849
Authority: WO
Inventors: Kaho Oyama; Kazumitsu Otsubo; Suguru Otsuka
Original assignee: Meiji Milk Products Co., Ltd.; Japan Science And Technology Corporation
Priority date: 1999-06-14
Filing date: 2000-06-14
Publication date: 2000-12-21
Also published as: AU5246900A

Abstract

Anticancer emulsions characterized by containing: (A) an anthracycline compound; (B) one or more saccharides in an amount of at least 10 w/v% in the aqueous phase; and (C) fats and/or a surfactant. The emulsion stability of the anticancer emulsions containing anthracycline compounds and the chemical stability of the anthracycline compounds in these emulsions are improved.

Description

Description Anticancer emulsion containing phosphorus compounds Technical field

The present invention relates to an anti-cancer emulsion containing an anthracycline-based compound having excellent emulsion stability and chemical stability. Background art

Recently, iodinated poppy oil fatty acid ester (“Lipiodol Ultra-Fluidj Laboratoire Gerbe”), an oil-based X-ray contrast agent, has been used for hepatic artery embolization, utilizing its ability to specifically deposit against liver cancer. A method has been used to achieve efficient delivery of anticancer drugs to liver cancer by using SMANCS, which is a covalent bond of styrene maleic acid copolymer (SMA) and water-soluble anticancer drug neocarnostin (NCS), for example. Oil-based preparations suspended and dissolved in Lipiodol (Hiroshi Maeda Supplement I 1998; 2: U9) and preparations in which Lipiodol and a water-soluble anticancer drug dissolved in a water-soluble X-ray contrast agent are emulsified (Kanematsu, T. et. Al. .: J. Surg. Oncol., 25: 218-226, 1984).

Among water-soluble anticancer drugs, anthracycline compounds show excellent antitumor effects, but have the problems of toxicity and low persistence of antitumor effects. Attempts have been made to overcome the point.

Examples of emulsions containing anthracycline compounds include doxorubicin hydrochloride and epirubicin hydrochloride as lipiodol emulsions, which are used for the treatment of local cancers such as liver cancer (Satyaj itBhattacharya et. Al. Cancer 1995; 76: 2202 -2210) is known. After administration, these anthracycline-based compound emulsions stay around the tumor and release the anthracycline-based compound to the lesion for a long period of time. Thus, it is characterized by exerting a therapeutic effect (Sun Woo Yi et. Al. Journal of Controlled Release 1998; 50: 135-143).

Anticancer emulsions containing these anthracycline compounds have the problem that the compounds are insolubilized in the aqueous phase and the chemical stability that they are susceptible to degradation such as hydrolysis. Emulsions, on the other hand, differ from aqueous solutions and solubilization systems in that they are thermodynamically unstable systems with dispersed particles. Therefore, in the case of an anticancer emulsion containing an anthracycline compound, both the chemical stability and the emulsion stability of the anthracycline compound must be satisfied in order to enable long-term release of the anthracycline compound. An important issue is the development of anticancer emulsions.

Until now, as an attempt to improve the emulsion stability of a general emulsion composition, an organic acid and a salt thereof (Japanese Patent Application Laid-Open No. 9-087162, Japanese Patent Application Laid-Open No. ), Salts (Jerzy Kizling et. Al. Colloids and Surfaces 1990; 50: 131-140), and sugars (Japanese Patent Laid-Open No. 6-287110). I have. However, even if organic salts such as sodium ascorbate and sodium citrate are added, and inorganic salts such as sodium chloride, magnesium chloride, and sodium sulfite are added, the anthracycline-based compound itself becomes water in the presence of the salt. Since the compound has the property of being hardly dissolved in water, it causes the compound to be insoluble, and a stable and uniform emulsion cannot be obtained. The addition of an acid or the like may not only cause local irritation at the time of administration by injection or the like, but also hydrolyze oil phase components and ester surfactant components in the prepared emulsion. As a result, problems such as a decrease in emulsion stability occur.

Further, it is said that the chemical stability of an anthracycline compound in a solution can be improved by adjusting the pH of the solution to 2.5 to 5.0 using an acid.

(Tokuhei 6—5 5667). However, as a result, problems such as a decrease in emulsification stability arise for the same reason as described above.

Accordingly, the emulsion stability of the anticancer emulsion containing an anthracycline compound, and It has been desired to develop an anticancer emulsion in which both the chemical stability of the anthracycline compound and the chemical stability of the emulsion are improved. Disclosure of the invention

In order to solve the above-mentioned problems, the present inventors systematically conducted research on a number of additives that are generally known to improve emulsion stability. It has been found that the above problems can be solved by including the saccharide of the present invention, and the present invention has been completed.

That is, the present invention provides (A) an anthracycline compound, (B) one or more saccharides in an aqueous phase of 10 w Zv% or more, and (C) fats and oils and Z or a surfactant. An anti-cancer emulsion characterized by comprising: BEST MODE FOR CARRYING OUT THE INVENTION

(A) The anthracycline compound is an active ingredient of the anticancer emulsion of the present invention, and is not particularly limited as long as it is an anthracycline anticancer compound. And doxorubicin hydrochloride, pirarubicin hydrochloride and the like, which are merely examples, and new anthracycline compounds which will be discovered in the future are also included in the present invention. Those skilled in the art can appropriately set the content of the compound according to the site, size, and the like of the cancer tissue. These anthracycline compounds are water-soluble and are usually present in the aqueous phase.

The (B) saccharides contained in the aqueous phase include sucrose, trehalose, maltose, glucose, fructose, monosaccharides such as xylitol, sorbitol, mannitol, disaccharides, sugar alcohols, and the like. This is merely an example, and the present invention is not limited to these saccharides. Up to one of these sugars Or two or more of them can be used in an appropriate combination. The aqueous phase contains 10 wZv% or more, preferably 20 wZv% or more, and more preferably 20 to 40 w / v%. In the aqueous phase, in addition to these, components such as amino acids can be reinforced as needed. Here, examples of the amino acid include L-aspartic acid and the like.

On the other hand, (C) fats and oils used in the oil phase include oxidized poppy oil fatty acid ester, medium-chain fatty acid triglyceride; vegetable oils such as soybean oil, sesame oil, castor oil, and poppy oil; and oleic acid. Higher fatty acids; benzyl alcohol, higher alcohols and the like can be mentioned, but these are only examples, and the present invention is not limited to these fats and oils.

The volume ratio of the water phase to the oil phase is preferably in the range of 1: 9 to 1: 0.2.

(C) Surfactants include polyoxyethylene hydrogenated castor oil, polyoxyethylene castor oil, polyoxyethylene sorbin monooleate (polysorbates), sorbin sesquioleate (spans), lecithin, etc. Examples of the nonionic surfactant include, but are not limited to, these surfactants. If necessary, they can be used alone or in combination of two or more. The addition amount of the surfactant can be appropriately set by those skilled in the art depending on the fats and oils used and the amount thereof, but is usually 0.1 to 20% by weight relative to the oil phase, particularly 0.5 to 1% by weight. It is preferably 0% by weight.

Emulsion has at least three components: an internal phase (dispersed phase), an external phase (dispersed medium), and an emulsifier, and is usually a liquid-liquid dispersion system in which water and oil are not mixed. Emulsions include oil-in-water (〇ZW) emulsions in which oil is the internal phase and water is the external phase, water-in-oil (WZO) emulsions in which water is the internal phase, and oil is the external phase.

ZO There are WZOZW type emulsion in which water type emulsion is dispersed in water, and OZWZO type emulsion in which OZW type emulsion is dispersed in water. The form of the emulsion of the present invention is preferably a WZO type emulsion or a WZOZW type emulsion, and may be a form of a mixture of an aqueous phase and an oil phase. An emulsion is composed of two or more types of phases that are not mixed with each other, by combining the composition and the manufacturing method, the interfacial properties (interfacial tension, viscoelasticity, etc.), It is a dispersion system stabilized by changing other physicochemical properties. The compositions shown here are the internal phase, external phase, emulsifier, emulsifying aid, presence or absence, type, composition ratio, etc. of other additives, etc. The order of addition of additives, mixing method, preparation temperature, equipment used, etc. Thus, many emulsifying methods are known because of the large number of factors involved in the formation of emulsions. Examples include the water-in-water emulsion method, the oil-in-water emulsion method (Jin Sezaki, Satoshi Kimura, "Pharmaceutics", Hirokawa Shoten, 1989), phase inversion emulsification, D-phase emulsification, amino acid gel Emulsification method (Hiromichi Uguisudani, Toshiyuki Suzuki, "Functions and applied technologies of emulsification and dispersion process", Science Forum, 1989), phase inversion temperature emulsification method, self-emulsification method, electroemulsification method (Kunio Furusawa, Fumio Kitahara) , Dispersion / Emulsification Chemistry ", Engineering Books, 197 5) etc. Machines and equipment for manufacturing emulsions are combined with the emulsification method described above, and the formulation and production of the desired emulsions Select in consideration of quantity.

The following are examples of emulsion production machines (Masasayoshi Ariga, Takayoshi Hirano, "Development of Pharmaceuticals 11, Pharmaceutical Unit Operations and Machinery", Yoshinobu Nakai, Hirokawa Shoten, P. 247-286, 1989).

1) A method using a shear force due to intense turbulence and / or the impact of a high-speed rotating blade, such as a homomixer. 2) A high-speed rotating grindstone, such as a colloid mill. 3) Method of impact by high-pressure fluid collision using high pressure difference, like Gorin Homogenizer 1) 4) Ultrasonic like homogenizer or siren type homogenizer 5) a method of dividing the liquid flow into multiple stages, and 6) a composite type combining multiple models and mechanisms. In addition, an emulsification method using a shear force generated when a pore is passed under pressure, such as a syringe pumping method, is widely used in in-hospital preparations and the like. The anticancer emulsion of the present invention can be prepared by appropriately combining the above emulsification method and emulsion production machine using the above components (A), (B) and (C).

For example, the WZO-type anticancer emulsion of the present invention can be prepared as follows. An anthracycline compound and saccharide are added to the purified water, mixed and dissolved with a homomixer, etc., and then an oil phase component and a surfactant are added thereto to disperse the water phase into the oil phase. can get.

The method may be in accordance with the above emulsification method. For example, a stirrer, a homogenizer, a high-speed homomixer ultrasonic machine or the like can be used.

The WZOZW-type anticancer emulsion of the present invention can be obtained, for example, by dispersing the WZ〇-type anticancer emulsion obtained above in an external aqueous phase. The outer water phase may contain other components. For example, if necessary, an aqueous surfactant can be contained. For example, polyoxyethylene hydrogenated castor oil, a block polymer type nonionic surfactant (Pull One-Pick Nick F 68 (“Poroxamer 188” manufactured by Midori Cross Co., Ltd.), etc.) can be used. The ratio is usually in the range of 1:10 to 1: 0.1, preferably in the range of 1: 5 to 1: 1.The volume ratio of the internal water phase to the oil phase is usually 1: 9 to 1: 0.1, preferably 1: 9 to 1: 0.5 These volume ratios can be appropriately set according to the type of the aqueous phase / oil phase, the site and properties of the cancerous tissue, and the like. . Example

Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited to these examples.

Production Example 1 Preparation of doxorubicin hydrochloride-lipiodol emulsion with maltose

As shown in Table 1, 2 mL each of maltose-containing aqueous solutions at various concentrations (1.25%, 2.5%, 5%, 10%, 20%, and 40%) and 2 m of purified water as a control were used. For reference, water-soluble contrast agent lopamiron 2mL (Nippon Schelling Co., Ltd.) 20 mg of a doxorubicin hydrochloride tracycline compound (manufactured by Sigma) was dissolved to obtain an aqueous phase (about 2 mL).

On the other hand, in response to each of the above, 80 mg of polyoxyethylene hydrogenated castor oil (Nikko Chemicals) with 50 moles of ethylene oxide was dissolved in 3.9 mL of Lipiodo (Laboratoire 'Gerbe), and the oil phase ( About 4 mL).

Next, while adding the above aqueous phase to each of the above oil phases in a high-speed homomixer (ULTRA-TURRAX T25, manufactured by IKA Laboratory), the aqueous phase was stirred for about 5 minutes to disperse the aqueous phase into the oil phase. A cancer emulsion was obtained.

Table 1 Oil phase Approx. 4 mL Water phase 2 mL

Lipio Doll Ho. Lioxyethylene maltose aqueous solution concentration

Hardened castor oil (W / V%)

Formula 1 3.9 mL 80 mg 1.25 ¾ 20 mg Formula 2 3.9 mL 80 mg 2.5 ¾ 20 mg Formula 3 3.9 mL 80 mg 5 ¾ 0 mg Formula 4 3.9 mL 80 mg 10% 20 mg Formula 5 3.9 mL 80 mg 20¾ 20 mg Formula 6 3.9 mL 80 mg 40% 20 mg Control 3.9 mL 80mg Purified water 20mg Reference example 3.9mL 80mg Iopamiron 300 20mg

Test example 1 Stability evaluation

1) Emulsion stability test

Each of the above emulsions was allowed to stand at room temperature for 72 hours after preparation, and its appearance was observed. That is, emulsification stability was evaluated by visually observing the occurrence of a heterogeneous phase.

2) Chemical stability test of doxorubicin hydrochloride and epilubicin hydrochloride

The aqueous phase component prepared from each of the above emulsions was sterilized through a 0.22 sterile filter, stored in a sterile tube at 55 ° C for 6 days in the dark and protected from light. Stability was evaluated by determining the residual ratio of the contained doxorubicin hydrochloride and epilubicin hydrochloride. The concentrations of epilubicin hydrochloride and doxorubicin hydrochloride contained in the aqueous phase component after storage were measured using the HPLC test method described below. The measurement conditions of the HPLC test method used are as follows.

Column) 0DS 5 m (4.6 X 250mm)

Mobile phase) Water-acetonitrile mixed solution (69:31, adjusted to pH 2.0 with concentrated phosphoric acid) Flow rate) About 0.8 mL / min

Temperature) Room temperature

Measurement wavelength) 254 nm

Standard solution) Approximately 25 mg of doxorubicin hydrochloride or epilubicin hydrochloride standard was precisely mixed and dissolved in the mobile phase to a concentration of about 0.12 mgZmL. This was filtered through a filter and used as a standard solution.

(Sample solution) Take an appropriate amount of each aqueous phase, add the mobile phase to it, dilute it to a concentration of about 0.1 mgZmL or less, and filter with a filter to obtain a sample solution.

Injection amount) 20 ^ 1

Calculation of concentration) It was calculated using the following equation.

Calculated concentration (mg / mL) = standard solution concentration (mg / mL) x AT / AS X dilution factor of sample solution

AS: Peak area of doxorubicin hydrochloride or epirubicin hydrochloride in standard solution AT: Peak area of doxorubicin hydrochloride or epirubicin hydrochloride in sample solution

3) Test results

Table 2 summarizes the results of the above emulsion stability test and chemical stability test. Table 2 Test method 1 (emulsion stability) Test method 2 (chemical stability) Additives

Appearance observation result Residual rate after 6 days at 50 ° C

(W / V%)

(Length of heterogeneous phase _mm ) ()

Control example No additive Heterogeneous phase (7.5) 80.7¾

Formula 1 Maltosed.25¾) With heterogeneous phase (7.0) 84.7%

Formulation 2 Maltose (2.5%) with heterogeneous phase (5.0) 84.9¾

Formulation 3 Maltose (5¾) with heterogeneous phase (3.5) 85.8%

Formulation 4 Maltose (10¾) Slightly heterogeneous phase 85.8%

Formula 5 Maltose (20%) homogeneous phase 85.1 8

Formula 6 Maltose (40¾) homogeneous phase 83.9%

Reference example Iopamiron300 homogeneous phase 77.4%

When the liquid is allowed to stand still, the dispersed phase may float to the liquid surface as a droplet or settle to the bottom, forming a layer. This phenomenon is called creaming. As is evident from Table 2, when the maltose content of the aqueous phase is 2.5% or less, the thickness of the heterogeneous phase formed by the creaming is as large as 5.0 to 7.0, which is a comparative example of 7. It is almost the same as 5 and has extremely poor stability. At a maltose content of 5.0%, it was improved to 3.5, or about 1Z2.At a maltose content of 10%, a heterogeneous phase was slightly observed. As in the above, no heterogeneous phase is observed. That is, this test revealed that emulsification stability was markedly improved by including maltose in the aqueous phase at 10% or more.

On the other hand, regarding the chemical stability of doxorubicin hydrochloride in the emulsion, the survival rate of the maltose-added group (1.25 to 20%) was 84.7 to 85.8%, which was 80% of the control. A clear improvement compared to 7% is observed. In the reference example, the residual ratio was 77.4%, which was worse than the control example, and did not correlate with the improvement in emulsion stability.

From these results, it is possible to improve both the emulsification stability of the anticancer emulsion containing an anthracycline compound and the chemical stability of the compound by including maltose in the aqueous phase at 10% or more. Became clear.

Test Example 2 Stability by adding various sugars Epilubicin hydrochloride was used as the anthracycline-based compound, and the aqueous phase contained the various sugars shown in Table 3 (maltose, sucrose, trehalose, glucose, fructose, sorbitol, and xylitol) at a concentration of 20%. Except for the above, according to Production Example 1, various sugar-containing epilubicin hydrochloride Z-lipiodol emulsion emulsions were prepared.

Table 3 Oil phase Approx. 4 mL Aqueous phase 2 mL Lipiodol e. Lioxyethylene epirvinno hydrochloride

Hardened 1: 7 oil 50

Formulation 7 3.9mL 80mg 20% Maltose 20mg Formulation 8 3.9mL 80mg 20 Sucrose 20mg Formulation 9 3.9mL 80mg 20% Trehalose 20mg Formulation 10 3.9inL 80mg 20 Glucose 20mg Formulation 11 3.9mL 80mg 20% Frac 1 20 mg formulation 12 3.9 mL 80 mg 20 Sorubi 1 ^ 20 mg formulation 13 3.9 mL 80 mg 20% xylitol 20 mg Control 3.9.9 mL 80 mg Purified water 20 mg As in Test Example 1, emulsification stability test, emulsion Epilubicin hydrochloride chemical stability test was performed. The results are summarized in Table 4.

Table 4 Test method 1 (emulsion stability) Test method 2 (chemical stability) Additives

Appearance observation result Residual rate after 6 days at 50 ° C

(W / V¾)

(Length of heterogeneous phase MI) (%)

Control example No addition 73.6%

Formulation 7 20% circle! 80.4%

Formulation 8 20% Sucrose 81.3%

Formula 9 20% Trehalose Proportion Proportion Proportion Proportion 81.2%

Formulation 10 20% glucose uniform knee 120.5¾

Nene Nene Nene.

Formulation 11 20% fructose eyes eyes eyes eyes 77.9%

Formulation 12 20% Sorbitol 79.8%

Formulation 13 20% Xylitol 83.0%

7

Five

As is clear from the table, by adding 2 OwZv% of maltose, sucrose, trehalose, glucose, fructoose, sorbitol, and xylitol in the aqueous phase, creaming like the control example was achieved in all sugars. No formation was observed, and improvement in emulsion stability was confirmed regardless of the type of sugar. On the other hand, regarding the chemical stability of epirubicin hydrochloride in the emulsion, the residual ratio was 77.9 to 83.0%, and there was no significant difference in the residual ratio depending on the type of sugar. It is clearly improved compared to 73.6%.

Industrial applicability

In preparing an anti-cancer emulsion containing an anthracycline compound, an aqueous phase contains an anthracycline compound and at least 10% (W / V) of at least one sugar, and an oil phase contains fats and oils and Z or parent compound. The inclusion of an oily surfactant markedly improved the emulsion stability of the anticancer emulsion and the chemical stability of the anticancer agent.

Claims

The scope of the claims

1. It contains (A) anthracycline compound, (B) 10 wZv% or more of one or more saccharides in an aqueous phase, and (C) fats and oils and Z or a surfactant. Anticancer emulsion.

2. The anticancer emulsion according to claim 1, wherein (A) the anthracycline compound is doxorubicin hydrochloride or epilubicin hydrochloride.

3. The anticancer emulsion according to claim 1, wherein (B) the saccharide contains 2 Ow / v% or more in an aqueous phase.

4. The anticancer emulsion according to any one of claims 1 to 3, wherein the emulsion is of a W / O type.

5. The anticancer emulsion according to any one of claims 1 to 3, wherein the emulsion is in the form of W / OZW.

6. The anticancer emulsion according to any one of claims 1 to 5, wherein the fat or oil is an iodinated coconut oil fatty acid ethyl ester.