JP2512310B2 - Coated fat emulsion - Google Patents

Description

TECHNICAL FIELD The present invention relates to a fat emulsion (hereinafter referred to as the fat emulsion of the present invention) in which fat emulsion particles are coated with a naturally-occurring polysaccharide derivative (hereinafter referred to as the derivative of the present invention).

2. Description of the Related Art Conventional problems and problems In the field of pharmaceuticals, in vivo use by injection administration of fat emulsions, that is, fat emulsions, has recently attracted attention. That is, conventionally, an emulsified system has been generally used as a dosage form for oral administration of an oily substance or as a dosage form for topical administration to the skin. However, it has become known that the emulsification system is also useful in so-called parenteral drug delivery systems. Therefore, from this aspect, an attempt has been made to use a dosage form intended to enhance the bioavailability in injection administration, and a dosage form different from the liposome. The following documents are shown as a reference for explaining such a tendency in detail.

SSDavis: Emulsion systems for the delivery of d
rugs by the parenteral route.Optimization of drug
delivery, Alfed Benzon Symposium 17.Editors; Hans Bu
ndgaard et, al. Copenhagen 1982. Now, a particularly problematic point in fat emulsions is that a technique for arbitrarily controlling the directivity of a drug to a so-called target organ has not yet been established. In addition, since a lipid emulsion generally has a high rate of elimination of a drug from the blood, it is difficult to maintain a constant blood concentration for a certain time. For this reason, a fat emulsion requires a so-called delay technique for reducing the disappearance rate, which has not yet been established.

Solution As a result of various studies to solve the above problems in fat emulsions, it was found that the object can be achieved by coating the particles of a fat emulsion with the naturally-occurring polysaccharide derivative according to the present invention, The present invention has been completed. That is,
In the fat emulsion administration, the lipid emulsion particles are coated with the naturally-occurring polysaccharide derivative according to the present invention for the purpose of controlling the directivity of the drug to the target organ and delaying the elimination of the drug into the blood. The subject is a fat emulsion characterized by.

 Hereinafter, the present invention will be described in detail.

In the present invention, fat emulsion means vegetable oil and animal oil 1 to 30 w / v%, emulsifier (phospholipid surfactant etc.) 0.1 to
It is mainly composed of 10 w / v%, an isotonicity agent, a pH adjusting agent and an appropriate amount of water.

Other fatty acids 2w / v% or less, cholesterol 3W / V%
Hereinafter, phosphatidic acid 2 W / V% or less, dicetyl phosphate 1 W / V% or less can be added.

The vegetable oil used in the present invention is not limited as long as it can be used for food or medicine, but soybean oil, sesame oil, cottonseed oil, olive oil and the like are preferable. In addition, eicosapentaenoic acid or the like is used as the animal oil.

Phospholipids are lecithins derived from egg yolk and soybeans, hydrogenated lecithin obtained by hydrogenating these (iodination degree 0 to 70).
Can also be used.

Surfactants are mainly nonionic surfactants,
Polyoxyethylene hydrogenated castor oil derivatives such as HCO-4
0, HCO-50, HCO-60, and polyoxyethylene polyoxypropylene ether derivatives such as Pluronic F-68 are mainly used.

Further, in the present invention, a phospholipid and a surfactant can be mixed and used as an emulsifier.

As the isotonicity agent, glycerin, glucose, malcase and the like can be used.

As the pH adjuster, hydrochloric acid, sodium hydroxide, trishydroxymethylaminomethane or the like is used.

The fatty acid may be linear or branched having 10 to 22 carbon atoms, and can be used as long as it can be used for food and medicine.For example, stearic acid, palmitic acid, myristic acid, oleic acid, linoleic acid. Are used. In addition, salts of these fatty acids can also be used, such as sodium salts,
A potassium salt or the like can be used.

The fat emulsion in the present invention has a particle size of 1 μm or less, preferably 0.2 to 0.4 μm.

Next, the naturally occurring polysaccharide derivative according to the present invention will be described.
It is as follows.

First, naturally occurring polysaccharides are pullulan, amylose, amylopectin, dextran, and mannan. One of the derivatives according to the present invention is, in these polysaccharides, 0.5 to 5.0 sugar units per 100 sugar units constituting the polysaccharide.
The primary hydroxyl group at the position carbon is represented by the formula —OCH ₂ CONHCH ₂ CH ₂ NHR ₁ (wherein R ₁ represents H or a cholesteryloxycarbonyl group). So, for example, in amylose,
0.5 to 5.0 sugar units per 100 It is shown like. Further in this derivative, R ₁
The sugar unit in which is a cholesteryloxycarbonyl group is 0.5
~ 4.5 pieces. The cholesteryloxycarbonyl group is represented by the following structural formula.

Another one of the derivatives according to the present invention is that 0.5 to 10.0 sugar units per 100 sugar units constituting the polysaccharide have a primary hydroxyl group at the 6-position carbon of the formula —OR ₂ (wherein R ₂ represents a straight chain acyl group having 12 to 20 carbon atoms), and a palmitic acid is particularly preferable.

Regarding the above-mentioned derivatives, there are JP-A-61-69801 and JP-A-58-201711, and the description therein is referred to. These derivatives are known to be used for coating the surface of liposomes, but they are not known to be used for fat emulsions as shown in the present invention.

 The fat emulsion of the present invention is produced as follows.

That is, a predetermined amount of vegetable oil or animal oil, an emulsifier,
An isotonicity agent, a lipophilic drug and other additives are mixed and heated, and an appropriate amount of water is added to the mixture to coarsely emulsify using a homomixer. Next, a pressure injection type homogenizer (Manton-Gaulin homogenizer) or an ultrasonic homogenizer is used to finely emulsify and obtain a homogeneous fat emulsion.

The naturally-occurring polysaccharide derivative-coated fat emulsion is obtained by mixing the fat emulsion thus obtained with the naturally-occurring polysaccharide derivative and stirring with a stirrer or ultrasonic treatment. The naturally-occurring polysaccharide derivative-coated fat emulsion of the present invention has an average particle size of 1.0 μm.
It is extremely fine as follows.

Whether the fat emulsion particles are coated with the derivative according to the present invention can be confirmed by the aggregation of the fat emulsion particles with concanavalin A, which is a type of lectin.

That is, concanavalin A has a property of binding to mannose and glucose and has a plurality of binding sites. Therefore, if the fat emulsion is coated with the naturally-occurring polysaccharide derivative, concanavalin A will be bound to the naturally-occurring polysaccharide derivative, and the fat emulsions will aggregate. On the other hand, if the fat emulsion is not coated with the naturally-occurring polysaccharide derivative, the aggregation of the fat emulsion by concanavalin A does not occur. Details of these will be described in Experimental Examples.

The drug compounded in the fat emulsion is a lipophilic substance,
Specifically, it is a pharmaceutical agent having an affinity for vegetable oils and animal oils, for example, ubidecarenone, tocopherol, tocopheryl acetate, tocopheryl nicotinate, vitamin K ₁ , vitamin K ₂ and air broth (PG) in a fat emulsion.
I ₂ ) and other plus-tagrandein, indomethacin fualnesol ester and the like can be mentioned.

Further, the naturally-occurring polysaccharide derivative according to the present invention to be incorporated in the fat emulsion is preferably 1 mg to 50 per 1 ml of the fat emulsion.
It is mg, but there is no particular limitation.

Action and effect The action and effect of the present invention can control the directivity of a drug to a target organ by appropriately selecting and coating the type of naturally-occurring polysaccharide derivative according to the present invention. The point is that the elimination rate of the drug can be delayed.

Examples Example 1 Purified egg yolk lecithin was added to 400 ml of water containing 12.5 g of glycerin.
6g was dispersed at 60-90 ° C. 60-90 beforehand in this solution
While gradually adding 50 g of soybean oil heated to ℃, it was emulsified with a homomixer for 30 minutes to obtain a crude emulsion. Then, add purified water to make 500 ml, and emulsify this liquid with a Manton-Gorlin homogenizer (first stage 100 kg / cm ² , total pressure 500 kg / c
m ² ) to obtain an extremely fine fat emulsion. The average grain size of this emulsion was 0.2 to 0.4 μm and contained no grains of 1 μm or more.

To 10 ml of the fat emulsion thus obtained, 3 ml of a cholesterol group-introduced mannan aqueous solution (24 mg / ml) was added, and the mixture was gently sonicated to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion.
The average particle size of this naturally occurring polysaccharide derivative-coated fat emulsion is 0.
It was 2 to 0.4 μm.

Example 2 To 50 g of soybean oil, 6 g of purified egg yolk lecithin, 0.15 g of sodium oleate and 0.15 g of phosphatidic acid were added, and the mixture amount was 60 to 90.
It melt | dissolved by heating at 0 degreeC. To this, add 400 ml of purified water preheated to 60-90 ° C, and then glycerin 12.
5 g was added, and purified water was further added to bring the total amount to 500 ml, and the mixture was roughly emulsified for 30 minutes with a homomixer. This was emulsified with a Manton-Gorlin homogenizer (first stage 100 kg / cm ² , total pressure 500 kg / cm ² ) to obtain an extremely fine fat emulsion. The average grain size of this emulsion was 0.2 to 0.4 μm and contained no grains of 1 μm or more.

To 10 ml of the fat emulsion thus obtained, 3 ml of cholesterol group-introduced amylopectin (60 mg / ml) was added and gently stirred with a stirrer to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion. The naturally-occurring polysaccharide derivative-coated fat emulsion had an average particle size of 0.2 to 0.4 μm.

Example 3 An uncoated fat emulsion was obtained in the same manner as in Example 1 except that 0.3 g of cholesterol was added to soybean oil in advance.
The average particle size was 0.2 to 0.4 μm, and particles having a size of 1 μm or more were not contained.

To 10 ml of the fat emulsion thus obtained, 3 ml of a cholesterol group-introduced pullulan aqueous solution (60 mg / ml) was added and gently sonicated to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion.
The average particle size of this naturally occurring polysaccharide derivative-coated fat emulsion is 0.
It was 2 to 0.4 μm.

Example 4 A CoQ ₁₀ -containing fat emulsion was obtained in the same manner as in Example 1 except that 1.25 g of CoQ ₁₀ was added to soybean oil in advance. . The average particle size was 0.2 to 0.4 μm, and the particles contained no more than 1 μm.

To 10 ml of the fat emulsion thus obtained, 3 ml of a cholesterol group-introduced mannan aqueous solution (24 mg / ml) was added, and the mixture was gently sonicated to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion.

The CoQ _10- containing naturally-occurring polysaccharide derivative-coated fat emulsion had an average particle size of 0.2 to 0.4 μm.

Example 5 An α-tocopherol-containing fat emulsion was obtained in the same manner as in Example 1 except that 5.0 g of α-tocopherol was previously added to soybean oil. The average particle size is 0.2-0.4 μm, 1
It did not contain particles of μm or larger.

To 10 ml of the fat emulsion thus obtained, 3 ml of a cholesterol group-introduced mannan aqueous solution (24 mg / ml) was added, and the mixture was gently sonicated to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion.
The CoQ _10- containing naturally-occurring polysaccharide derivative-coated fat emulsion had an average particle size of 0.2 to 0.4 μm.

Example 6 An ailoprost-containing fat emulsion was obtained in the same manner as in Example 1 except that 5.0 mg of ailoprost was previously added to soybean oil. The average particle size was 0.2 to 0.4 μm and contained no particles of 1 μm or more.

To 10 ml of the fat emulsion thus obtained, 3 ml of a cholesterol group-introduced mannan aqueous solution (24 mg / ml) was added, and the mixture was gently sonicated to obtain a naturally-occurring polysaccharide derivative-coated fat emulsion.
The average particle size of this ailoprost-containing naturally-occurring polysaccharide derivative-coated fat emulsion was 0.2 to 0.4 μm.

Experimental Example 1 Sample In the same manner as described in Example 1 and Examples 4 to 6, sample samples a-1 below were prepared.

Uncoated fat emulsion b. Naturally-derived polysaccharide derivative-coated fat emulsion, but naturally-derived polysaccharide derivative is cholesterol group-introduced mannan (molecular weight 200,000, cholesterol group substitution degree 2.3) c. Naturally-derived polysaccharide derivative-coated fat emulsion, but naturally-derived polysaccharide Derivatives are cholesterol group-introduced amylopectin (molecular weight 112,000, cholesterol group substitution degree 1.8) d. Naturally-derived polysaccharide derivative-coated fat emulsion, provided that naturally-occurring polysaccharide derivatives are palmitoyl group-introduced amylopectin (molecular weight 112,000, palmitoyl group substitution degree 8.0) e. Polysaccharide derivative-coated fat emulsion, but naturally-derived polysaccharide derivative is cholesterol group-introduced bullan (molecular weight 50,000, cholesterol group substitution degree 1.9) f. Ubidecarenone-containing naturally-derived polysaccharide derivative, but ubidecalenone-containing 2.5 mg / ml, naturally-derived Polysaccharide derivative is cholesterol group Input mannan (molecular weight 200,000, degree of cholesterol group substitution 2.3) g. Α-tocopherol-containing naturally-occurring polysaccharide derivative-coated fat emulsion, provided that α-tocopherol is contained at 10 mg / ml. Cholesterol group substitution degree 2.3) h. Α-tocopheryl acetate containing naturally-occurring polysaccharide derivative-coated fat emulsion, provided that α-tocopheryl acetate is 5 m
g / ml, naturally-derived polysaccharide derivative is cholesterol group-introduced pullulan (molecular weight 65,000, cholesterol group substitution degree 1.7) i. Vitamin K _1- containing naturally-occurring polysaccharide derivative-coated fat emulsion, but containing vitamin K ₁ 3 mg / ml, A naturally-occurring polysaccharide derivative is a cholesterol-introduced amylopectin (molecular weight 112,000, cholesterol group substitution degree 2.0) j. Vitamin K ₂ -containing naturally-occurring polysaccharide derivative-containing fat emulsion, provided that vitamin K ₂ is contained at 5 mg / ml. Group-introduced mannan (molecular weight 200,000, cholesterol group substitution degree 2.3) k. Iloprost-containing naturally-occurring polysaccharide derivative-coated fat emulsion, provided that 10 μg / ml of ailoprost is contained, and the naturally-occurring polysaccharide derivative is cholesterol-group-introduced mannan (molecular weight 20
0,000, degree of cholesterol group substitution 2.3) l. Naturally-derived polysaccharide derivative-containing fat emulsion containing indomethacin-farnesol ester, provided that indomethacin-farnesol ester contains 10 mg / ml and the naturally-occurring polysaccharide derivative has cholesterol group-introduced mannan (molecular weight 200,000). , Cholesterol group substitution degree 2.3) Method Measurement of average particle size The average particle size (diameter) of the fat emulsion before and after coating with the naturally-occurring polysaccharide derivative was measured by a submicron analyzer.

Confirmation of coating of naturally-occurring polysaccharide derivative on the surface of fat emulsion Samples a to l were diluted 1/1000 with 20 mM Tris-HCl buffer (pH 7.2), and concanavalin A, which is an agglutinin that recognizes sugar residues, was diluted with this. Was added and the naturally occurring polysaccharide derivative coating was assayed from the aggregation of the fat emulsion. Aggregation of the fat emulsion was examined by the change in absorbance at 620 nm.

Results Table 1 shows the average particle size of the naturally occurring polysaccharide derivative-coated fat emulsion.
Shown in

Description of Table 1 As shown in Table 1, there is no significant change in the average particle size of the fat emulsion before and after coating depending on the type and amount of the naturally-occurring polysaccharide derivative coated.

Concanavalin A, a naturally occurring polysaccharide derivative-coated fat emulsion
Aggregation by addition is shown in FIG.

Description of FIG. 1 1) to 5) are the following samples: 1); b-1 sample of Table 1 2); b-2 sample of Table 1 3); b-3 of Table 1 4); sample of a in Table 1 5); cholesterol group-introduced mannan only.

The arrow indicates the time point when Concanavalin A (250 μg / 0.1 ml buffer) was added to the measurement sample.

The horizontal axis of the figure represents time (minutes), and the vertical axis represents changes in absorbance at 620 nm. Abs · _t indicates the absorbance at 620 nm at time t, and Abs · ₀ indicates the absorbance at 620 nm when concanavalin A was not added.

As shown in FIG. 1, since the aggregating ability increases with the added amount of the coated naturally-derived polysaccharide derivative, the naturally-occurring polysaccharide derivative coated amount also increases with the added amount. The uncoated fat emulsion has no aggregating ability, and only a slight increase in absorbance was observed with the cholesterol group-introduced mannan.

Experimental Example 2 Sample A specimen sample of the following ac was prepared by the same method as in Example 4 except that ¹⁴ C-CoQ ₁₀ was used instead of CoQ _{10 in} the description of Example 4.

¹⁴ C-CoQ _10- containing fat emulsion b. ¹⁴ C-CoQ _10- containing naturally-occurring polysaccharide derivative-coated fat emulsion, provided that the naturally-occurring polysaccharide derivative is a cholesterol group-introduced mannan (molecular weight 200,000, cholesterol group substitution degree 2.3) Derivative coverage of 7.2 for 1 ml of fat emulsion
mg c. ¹⁴ C-CoQ ₁₀ containing naturally-derived polysaccharide derivative-coated fat emulsion, provided that naturally-occurring polysaccharide derivative is cholesterol group-introduced amylopectin (molecular weight 112,000, cholesterol-group substitution degree 1.8). 7.2
Note that ¹⁴ C-CoQ ₁₀ is radiolabeled ubidecarenone represented by the following structural formula.

Method Animal experiment Male guinea pigs (body weight 280-350 g) were injected with 0.76 mg ¹⁴ C-CoQ ₁₀ / kg of the sample sample into the left femoral vein and sutured. After that, the guinea pig was left in the breeding cage, and blood was collected from the ear vein every predetermined time. The radioactivity concentration in the collected blood was measured by the method described below.

Incidentally, 30 minutes and 24 hours after administration, blood was removed from the aorta under anesthesia with a syringe and killed, and each organ was excised.

Measurement of blood radioactivity 20 μl or 50 μl of blood was collected from the ear vein and 0.75
Soluene 350 / isopropyl alcohol (1/1, volume ratio) solubilized, and after adding a few drops of hydrogen peroxide water to decolorize,
5 ml of instagel / 0.5N HCl (9/1, volume ratio) was added, and the radioactivity was measured using a liquid scintillation counter.

Measurement of radioactivity in organs Approximately 30 mg of organ was added to 0.5 ml of Soluene350, at room temperature, approximately 12
After solubilizing the organs by incubating for an hour, 5 ml of ins
Tagel / 0.5N HCl (9/1, volume ratio) was added, and the radioactivity was measured using a liquid scintillation counter.

Results FIG. 2 shows changes in the radioactivity concentration in blood after the administration of the sample specimen.

FIG. 3 and FIG. 4 show the organ-to-organ transfer between samples at 30 minutes and 24 hours after administration.

Description of FIG. 2 The circles show the case where the sample a was administered, the circles show the case where the sample b was administered, and the triangles show the case when the sample c was administered (average value of 3 cases). Shown respectively.

From Figure 2 ¹⁴ C-Co in fat emulsion coated with naturally occurring polysaccharide derivatives
It can be seen that the elimination of Q _{10 from} the blood is slower in the early period after administration than that of ¹⁴ C-CoQ _{10 in} the uncoated fat emulsion.

Description of FIGS. 3 and 4 Each column of shows the amount of organ transfer when each of sample a, sample b, and sample c is administered.

Coating the fat emulsion with cholesterol-introduced mannan markedly increased pulmonary and hepatic translocation. On the other hand, the transition to the heart and adrenal gland was significantly reduced. In addition, the cholesterol group-introduced amylopectin-coated fat emulsion tended to increase migration to the liver.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the relationship between the added amount of the derivative and the aggregating ability in the naturally occurring polysaccharide derivative-coated fat emulsion of the present invention. FIG. 2 is a graph showing changes in the radioactivity concentration in blood after the administration of the sample specimen. FIG. 3 and FIG. 4 show the organ-to-organ transfer between samples at 30 minutes and 24 hours after administration, in contrast.

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Sumio Watanabe 9-6 Inarimae, Yatabe-cho, Tsukuba-gun, Ibaraki Vila Espoir 301 (72) Inventor Yasuo Miyake 2-20-5 Kasuga, Yatabe-cho, Tsukuba-gun, Ibaraki Tsukuba Heights 202 (72) Inventor Junzo Sunamoto 4-16-10 Yokoo, Nagasaki City (56) Reference JP-A-58-201711 (JP, A) JP-A-61-69801 (JP, A)

Claims (3)

(57) [Claims] 1. Pullulan, amylose, amylopectin,
In dextran and / or mannan, 0.5 to 5.0 sugar units per 100 sugar units constituting it are
The primary hydroxyl group at the 6-position carbon is represented by the formula --OCH ₂ CONHCH ₂ CH ₂ NHR ₁ (wherein R ₁ represents H or a cholesteryloxycarbonyl group), and R ₁ represents cholesteryloxycarbonyl. In the case of a naturally occurring polysaccharide derivative having 0.5 to 4.5 sugar units when it is a group, or pullulan, amylose, amylopectin, dextran and / or mannan, 0.5 to 10.0 sugar units per 100 sugar units constituting it are , A naturally occurring polysaccharide derivative whose primary hydroxyl group at the 6-position carbon is represented by the formula —OR ₂ (wherein R ₂ represents a straight-chain acyl group having 12 to 20 carbon atoms), is used to coat fat emulsion particles. Fat emulsion characterized by being. 2. A fat emulsion containing ubidecarenone, tocopherol, tocopheryl acetate, tocopheryl nicotinate, vitamin K ₁ , vitamin K ₂ , ailoprost and other prostaglandins, indomethacin fuarnesol ester. The fat emulsion according to claim 1, characterized in that 3. The fat emulsion according to claim 1 or 2, wherein the naturally-occurring polysaccharide derivative is 1 mg to 50 mg per 1 ml of the fat emulsion.