JPH0469146B2 - - Google Patents

Description

[Detailed description of the invention]

[Field of Application] The present invention relates to a benzoylurea compound/phospholipid complex. [Prior art] General formula (In the formula, X represents a halogen atom or a nitro group, Y and Z ₂ each represent a hydrogen atom or a halogen atom, Z ₁ represents a halogen atom or a trifluoromethyl group, and A represents a group =CH- or =N- ) It has already been disclosed that the benzoyl urea compound represented by
109721). This benzoyl urea compound has the property of being difficult to dissolve in water and organic solvents. In general, methods for formulating or solubilizing poorly soluble drugs include:
Examples include emulsification with surfactants, inclusion with cyclodextrins, derivatization, liposome formation, and microencapsulation. However, even with these measures, in the case of benzoylurea compounds,
It has been difficult to prepare preparations that can exhibit sufficient medicinal efficacy. Therefore, the present inventors conducted various studies on formulation of this benzoyl urea compound, and found that by utilizing the affinity between the benzoyl urea compound and phospholipid, and forming a complex between the benzoyl urea compound and the phospholipid, the benzoyl urea compound The present invention was completed based on the discovery that the complex has excellent stability and can be used as a pharmaceutical, and that it can also be administered intravenously by adjusting the particle size of the complex. [Disclosure of the Invention] The present invention relates to a benzoylurea compound/phospholipid complex. The benzoyl urea compound used in the present invention has the general formula (In the formula, X represents a halogen atom or a nitro group, Y and Z ₂ each represent a hydrogen atom or a halogen atom, Z ₁ represents a halogen atom or a trifluoromethyl group, and A represents a group =CH- or =N- ) is not particularly limited. The manufacturing method is also disclosed in the above-mentioned Japanese Patent Application Laid-Open No. 57-109721. As the phospholipid for forming the complex, any non-toxic phospholipid that is physiologically acceptable and metabolizable can be used in the present invention. For example, phosphatidylcholine, phosphatidylserine, phosphatidic acid, phosphatidylglycerin, phosphatidylethanolamine, phosphatidylinositol, sphingomyelin, dicetyl phosphate, lysophosphatidylcholine (lysolecithin), stearyl Amines or mixtures thereof such as soybean phospholipids and egg yolk phospholipids are used. Preferred phospholipids include:
Examples include phospholipids from soybeans or egg yolks. Both of these components are used to prepare the complex of the present invention. First, a phospholipid and a benzoylurea compound are dissolved in an organic solvent such as chloroform. The ratio of the benzoyl urea compound to 1 part by weight of the phospholipid is preferably 0.01 to 10 parts by weight. The container containing the solution in which the drug and phospholipid are mixed is preferably vacuumed using a rotary evaporator to remove the solvent, and the phospholipid is thinly adhered to the inner wall of the container to form a phospholipid containing a benzoyl urea compound. Form a thin film. In this case, preferably for the stabilization of the phospholipids, an antioxidant, such as α-tocopherol, is preferably added in a proportion by weight relative to the phospholipids.
Add so that the amount is about 0.0001 to 0.5% (w/w). It is also of course possible to add known stabilizers. The formed thin film is coated with a physiologically acceptable aqueous solution (e.g. PH5.5-8, preferably PH6-7).
adjusted to. Specifically, citrate buffer,
acetate buffer, phosphate buffer, physiological saline solution, etc.)
is added in an amount of 10 to 500 ml per 1 g of phospholipid, and immediately shaken or stirred to break the thin film and form particles consisting of a composite. Specifically, the usual method is to put several glass beads in a round-bottomed flask and shake them vigorously by hand (for 3 to 20 minutes at room temperature). A complex suspension with a concentration of 0.5 to 100 mg phospholipid/ml is thus prepared. The particles are then subjected to ultrasonic treatment to adjust the particle size to 0.2 to 2 μm or less. At this time, in order to prevent heat generation (increase in liquid temperature) due to ultrasonic waves, ultrasonic treatment is performed under ice cooling. Moreover, it is preferably carried out under an inert gas atmosphere such as nitrogen. Furthermore, centrifugation is carried out at a temperature of 5 to 20℃ for 10,000 g or more.
Preferably 20,000 to 30,000 g is used for about 30 minutes to 2 hours. As a result, the suspension separates into three layers. That is, the upper layer (referred to as Fr) is the supernatant that shows the color of phospholipids, and the upper layer (referred to as Fr) is the supernatant that shows the color of phospholipids.
unilamellarvesicle) worthy of fraction. Middle layer (Fr
) is a precipitate that shows a mixed color of drug and phospholipid, and is a layer of lipid particles that remain unbroken by sonication. Fr is further converted into Fr and Fr by ultrasonication. The bottom layer (referred to as Fr) of the sediment has a color depending on the drug. This bottom layer sediment Fr is collected and centrifugally washed using the above-mentioned physiologically acceptable aqueous solution, phospholipid-containing aqueous solution, or supernatant Fr. In this way, a poorly water-soluble drug/phospholipid complex is prepared in the form of a pellet or suspension. The complex obtained according to the present invention may be formulated according to methods widely known in pharmaceuticals. Furthermore, by freeze-drying the liquid preparation, it can also be provided as a dry preparation. In this case, it is of course free to use known stabilizers as appropriate depending on each drug or phospholipid. Additionally, adding a nonionic surfactant is useful for increasing the solubility of the dry formulation. Such dry preparations are generally used after being dissolved or diluted with a physiologically acceptable aqueous solution, such as a physiological saline solution or the above-mentioned supernatant Fr. encapsulation, enteric coating,
It may be made into a suspension, granule, powder, injection, suppository, etc. The complex obtained by the present invention has increased solubility in water compared to the zenzoyl urea compound itself,
With solubilization, the activity increases several to several tens of times. Furthermore, since the toxicity is suppressed to a lower level than when administered alone, it is expected that the efficacy will increase with increasing dosage. In addition, by adjusting the particle size of the complex to 0.2 to 2 μm, it is possible to make it into an injectable formulation, which makes it possible to administer it intravenously, which is expected to improve fast-acting or local affinity. It is expected to suppress the side effects of drugs on the intestinal tract, and at the same time increase efficacy by increasing intestinal absorption. The complex comprising the present invention thus provided is
It is possible to formulate poorly water-soluble drugs in an unprecedented and effective manner, and it will enable new developments in the medical industry and clinically. Experimental Example 1 The degree of incorporation of the benzoylurea compound (compound 1) into the phospholipid in each layer (Fr~) after centrifugation was investigated. Each layer was separated and treated with Triton The amount of drug per 100 mg of lipid was calculated (Table 1). Experimental Example 2 The size of particles in each layer (Fr~) was investigated by centrifugal sedimentation method (Coulter counter method) (Table 1).

[Table] Experimental Example 3 The in vivo effect of the benzoylurea compound/phospholipid complex obtained according to the present invention on P-388 leukemia cells was investigated using an ip-ip system. Specifically, P-388 leukemia cells were intraperitoneally transplanted into mice with CDF ₁ at a rate of 1 x 10 ⁶ cells/mouse, and the complex of the present invention was intraperitoneally administered on the 1st and 4th day after inoculation, and the cells were incubated for 30 days. The mice were observed to be alive or dead. Using physiological saline as a control, the effect of each drug was calculated and expressed as T/C, taking the effect as 100%.

[Table] Experimental example 4 1×10 ⁵ L-1210 leukemia cells were added to BDF ₁ mice.
The complexes of the present invention were intraperitoneally implanted at a ratio of 2 mice/mouse, and the complexes of the present invention were administered intravenously. The survival rate (%) of each treatment group (10 mice per group) was determined by observing the survival of the mice for 30 days and setting the number of survival days of mice in the control group to which physiological saline was administered as 100 (Table 3).

[Table] Experimental Example 5 The acute toxicity, dosage, and administration method of the complex obtained by the present invention were investigated. (Acute toxicity) In intravenous administration using CDF ₁ mice (10 mice), only 1 mouse died even after administering 500 mg/Kg.
Since there were no fish, the acute toxicity ( _LD50 ) was
It was confirmed that the amount was 500mg/Kg or more. (Dosage) Although the dosage cannot be determined unconditionally due to differences in administration conditions, it is usually about 5 to 50% per day for the active ingredient.
It is 1000mg/Kg. When administering the present complex, it is of course possible to administer the above-mentioned dosage all at once or in divided doses. (Administration method) The complex obtained by the present invention can be administered orally, intravenously,
It can be administered by methods such as intrarectally. Example 1 After dissolving 50 mg of compound 1 (see experimental example), 500 mg of purified egg yolk phospholipid, and 0.5 mg of α-tocopherol in 10 ml of chloroform, the mixture was heated under reduced pressure using a rotary evaporator to distill off the chloroform.
A thin film of phospholipid containing Compound 1 was formed.
Add 10 ml of physiological saline to this thin film, immediately shake vigorously at room temperature for 20 minutes, and then use a sonicator (Branson Sonic Power, Cell Disrutor).
#350, output 60W), ultrasonication was performed for 1 hour while cooling on ice. Further centrifugation (25,000g, 1 hour) was performed at room temperature, the bottom layer sediment was collected, centrifugally washed several times using the above-mentioned physiological saline, and sterilized and filtered to remove suspended phosphorus. A lipid complex was obtained. Example 2 The suspension preparation obtained in Example 1 was freeze-dried to obtain 520 mg of a dry preparation of a phospholipid complex. Example 3 The procedure of Example 1 was followed except that Compound 8 (25 mg) was used instead of Compound 1 (50 mg). After sterile filtration, albumin (final concentration 0.5%) was added, freeze-dried, and phosphate was removed. A dry formulation of lipid complex was obtained.

Claims (1)

[Claims] 1. General formula (In the formula, X represents a halogen atom or a nitro group, Y and Z ₂ each represent a hydrogen atom or a halogen atom, Z ₁ represents a halogen atom or a trifluoromethyl group, and A represents a group =CH- or =N- ) A benzoyl urea compound/phospholipid complex consisting of a benzoyl urea compound and phospholipid. 2. After dissolving the benzoyl urea compound and phospholipid in an organic solvent, the solvent is distilled off to form a lipid thin film, and the suspension of this thin film is further sonicated and then centrifuged to obtain the bottom layer precipitate. The benzoylurea compound/phospholipid complex according to claim 1, which is obtained by recovering the benzoyl urea compound/phospholipid complex.