JPH029816A - Supergualin preparation - Google Patents

Abstract

PURPOSE:To provide a supergualin preparation containing supergualins and complex lipids, applicable by oral administration, etc., other than injection and useful as a carcinostatic agent and/or an immunosuppression agent. CONSTITUTION:The objective preparation is obtained by mixing 1 pt.wt. of supergualins with 1-50 pts.wt. of a complex lipids (e.g., phosphatidylcholine), compounding conventional additives, solvents, etc., and forming in the form of drug preparation by conventional method. The amounts of the supergualins and the complex lipids are 0.015-90% and 0.015-98% based on the whole preparation, respectively. An example of the supergualins is N-[4-(3-aminopropyl) aminobutyl]-2-(7-guanidinoheptanamido)-2-hydroxyethanamide. High blood concentration of supergualins having poor absorbability can be attained even by the administration other than injection.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a novel preparation of spagarins that are expected to be used as anticancer agents and/or immunosuppressants.

[Conventional technology]

Spagarin is an antibiotic with interrupting activity discovered by Umezawa et al. (Japanese Patent Laid-Open No. 57-48957). Since then, many derivatives and related compounds have been found to have similar activity (Japanese Patent Laid-Open No. 57-48957). 58-62152, 59-
42356. 60-185758. 62-48
660, etc.), and it was also found that it has an immunosuppressive effect (Japanese Patent Application Laid-Open No. 129119/1989).

Some of these compounds are currently showing promise as anticancer agents and/or immunosuppressants. As for the preparation, an injection preparation has been developed (Japanese Patent Application Laid-Open No. 61-165322).

[Problems to be Solved by the Invention] Spagarin and its derivatives or related compounds (hereinafter collectively referred to as spagarins) exhibit anticancer effects and/or immunosuppressive effects when administered other than conventional injections. It was difficult to obtain sufficient blood concentrations.

However, in order to treat or prevent the target disease, it is necessary to administer spagarins continuously to some extent, and there is a desire to develop formulations that are easy to administer, such as by oral administration in addition to injection.

[Means for Solving the Problems] As a result of various studies, the present inventors found that there are 15 formulations containing spagarins and complex lipids (hereinafter simply referred to as lipids).
-deoxyspergarin (DSG) and N-(4-(3-aminopropyl)aminobutyl]2-(7-guanidinohebutanamide)-2-methoxyethanamide (MeDS).
We have completed the present invention by discovering that even in spagarins with poor absorption such as G), high blood concentrations can be obtained by administration routes other than injection, such as oral administration.

The spagarins in the present invention are not particularly limited; for example, general formula (I) general formula (1) % formula %) (in the formula + R1 is -(CHz)4-, -(C1h)b-
9-Q-CI(z-1Q-(CH2)Z-, -cH,-
represents Q-, and R1 is -CH,-.

-(C1h)! -1-CM(OH)CHz- or-
C1 (, CH-, R3 is -CH(OH)-, -C
H(OCHz)-, -CHz- or -CH(CH,0
H)-, and X represents a hydrogen atom or a group obtained by removing the hydroxyl group from the carboxylic acid of an amino acid or peptide,
) and their pharmacologically acceptable salts.

Specifically, the following compounds can be mentioned.

(1) N-(4-(3-aminopropyl)aminobutyl)-2-(7-guanidinohebutanamide)-2-hydroxyethanamide (DSG) (2) L (4-
(3-aminopropyl)aminobutyl)-2-(7-guanidinohebutanamide)2-methoxyethanamide
(MeDSG) (3) N-(4-(3-aminopropyl)aminobutyl)-2-(9-guanidinononanamide)-2=hydroxyethanamide (4) N-(4-(3-aminopropyl) N-(4-(3-aminopropyl)aminobutyl)-2-(7-guanidinohebutanamide)-(s)
-2-hydroxyethanamide (6) N-(4-(3-aminopropyl)aminobutyl) -2-(4-(ρ-guanidinophenyl)butanamide) ethanamide (7) N-(4-(3-amino (propyl)aminobutyl) -2-(4-(p-guanidinophenyl)butanamide)-(s)-2-hydroxymethylethanamide (8) N-(4-(3-aminopropyl)aminobutyl) -2- (3-(p-guanidinomethylphenyl)
propanamide)-(s)-2-hydroxymethylethanamide (9) N-(4-(3-aminopropyl)aminobutyl)-2-(5-(p-guanidinophenyl)pentanamide)-(s) -2-Hydroxymethylethanamide (10)N-(4-(3-aminopropyl)aminobutyl)-2-(7-guanidinohebbut-2-enamide)
-2-methoxyethanamide (11)N-(4-(3-aminopropyl)aminobutyl) -2-(9-guanidinonon-2-enamide)
-2-hydroxyethanamide (12) N-(4-(3-phenylglycylaminopropyl)aminobutyl) -2-(4-(p-guanidinophenyl)butanamide]-2-hydroxymethylethanamide (13) N-(4-(3-leucylleucylaminopropyl)aminobutyl]-2-(4-(p-guanidinophenyl)butanamide]-2-hydroxymethylethanamide (14) spagarin, and the like.

-JC The compound of formula (1) forms a salt with an acid.

The acid for forming a salt may be either an inorganic acid or an organic acid as long as it is pharmacologically acceptable. Examples of inorganic acids preferably include hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid; For example, acetic acid, propionic acid, succinic acid, fumaric acid.

Maleic acid, malic acid, tartaric acid, glutaric acid.

Citric acid, benzenesulfonic acid, toluenesulfonic acid,
Methanesulfonic acid, ethanesulfonic acid.

Propanesulfonic acid, asparagine H, ri)Ii9minic acid, etc. are preferred.

The lipids used in the present invention include phosphatidylcholine (PC) and phosphatidylethanolamine (PE).
, phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG
), glycerophospholipids such as phosphatidic acid (PA), sulflipids, glycolipids, sphingoids, ceramides, sphingolipids, and sphingophospholipids can be used. These lipids may be natural lipids such as egg yolk phospholipids or soybean phospholipids, or lipids obtained by hydrogenating or hydroxylating natural lipids, and/or synthetic lipids. It is okay to mix and use them depending on the purpose, and it is also okay to add simple lipids that are not included in complex lipids to change the properties of the lipid mixture or to stabilize it.1 For example, to stabilize lipids. Addition of vitamin E, etc. for measurement purposes, addition of cholesterol, etc. for the purpose of stabilizing lipid membranes, and gusetyl phosphate to impart charge to lipid membranes. Stearylamine etc. may be added.

The formulation of the present invention is characterized by containing spagarins and lipids, and is in liquid form.

Spagarins and lipids dissolved and dispersed in water, regardless of their solid form, take the form of what is commonly called a liposome, but it is not necessary that the spagarins be incorporated into liposome vesicles. Therefore, a preparation containing a mixture of lipids and spagarins can be made into a dosage form suitable for administration, regardless of the general method for preparing drug-loaded liposomes. When obtaining a mixture of spagarins and lipids using a liposome preparation method, general methods for preparing liposomes such as the thin film method, reversed phase inhalation method, surfactant dialysis removal method, ethanol injection method, and French press method are used. It will be done. These preparations are aqueous solutions containing lipids and spagarins and can also be administered as such. When the preparation is in a solid state, the above aqueous solution can be obtained as a powder by freeze-drying or spray-drying, and this powder can be used to form capsules of 1.
tablet.

It is also possible to formulate granules or powders.

In addition, the present invention requires that spagarins and fat f[ be mixed at the time of administration, and 1 does not necessarily require that spagarins and lipids be mixed in advance, so a lipid preparation and spagarins are administered. A formulation that is sometimes mixed may also be used; an example of this is a method in which a lyophilized lipid powder is dispersed in an aqueous solution of spagarin at the time of administration to prepare a mixture.

The mixing ratio of spagarins and lipids depends on the biological activity and absorbability of spagarins, but the upper limit is not particularly limited to 1 part (by weight) of fat to 1 part of spagarins (by weight), but the upper limit is not particularly limited, but depending on the formulation It can be decided accordingly. The amount of lipid mixed is preferably about 1 to 50 parts by weight per 1 part of spagarin.

The ratio of spagarin in this preparation is 0.0 to the total amount of the preparation.
The amount of lipid is about 15 to 90%, preferably about 0.1 to 50%, and the amount of lipid is about 0.015 to 98%.

Preferably it is about 0.1 to 98%.

In addition to spagarins and lipids, additives, solvents, etc. may be added to obtain the desired dosage form. In order to obtain a solution-like dosage form, a pharmacologically acceptable solvent is required. used. Generally, the solvent is water 1 ethanol,
Alcohols such as propylene glycol and mixed solvents thereof are used. Various additives can be added to these solvents depending on the purpose. For example, various pharmacologically acceptable inorganic salts and organic salts can be used as buffers. 1!
Examples include adding i as a flavoring agent and surfactants as emulsion stabilizers.

Various additives may be added for various purposes in order to obtain solid preparations.

The amount of these additives added depends on the target formulation, but
0-1000 for 1 part of mixture of spagarins and lipids
This is within the scope of the department.

Next, the effects of the present invention will be illustrated by experimental examples.

[Effect]

·experiment.

Mixtures of spagarins and lipids shown in Examples 1 to 3 as samples of the present invention and aqueous solutions of spagarins shown in Control Example 1 as a control were orally administered to rats, and their blood concentrations were compared.

- Experimental Method 8 The products of the present invention of Examples 1 to 3 and the control product of Control Example 1 were administered to F344 rats of appropriate age at a dose of 50% spagarin.
mg/kg was administered orally. Blood was collected over time.1. The concentration of spagarins in blood was measured by liquid chromatography.

-Results As is clear from the results shown in Figure 1, the three products of the present invention showed significantly higher blood concentrations and/or persistence than the control product. In addition, when calculating the blood concentration area from Figure 1, Table 1
It's like this.

Table 1. Area under blood concentration of MeDSG As is clear from this table, the area under blood concentration of the product of the present invention was 1.5 to 2 times or more than that of the control product.

Therefore, the absorption rate of the present invention is 1.5 to 2 times higher than that of the control product.

[Effects of the Invention] From the above description, the present invention can be expected to provide a preparation that exhibits the anticancer and immunosuppressive effects of spagarins by oral administration, etc., in addition to injection.

Hereinafter, the composition of component 1 and the preparation method of the mixture of spagarins and lipids of the present invention will be specifically described with reference to Examples.
The formulations of the present invention are not limited to these examples.

Example 1 Dipalmitoylphosphatidylcholine (DPPC)
670o+g, dioleoylphosphatidylcholine (DOPC) 440n+g, dipalmitoylphosphatidylserine (DPPS) 74+g, cholesterol (Chol) 96mg and N-(4-(3-
(aminopropyl)aminobutyl) -2-(7-guanidinohebutanamide)-2-methoxyethanamide (
A reverse phase liposome gel containing an aqueous solution of MeDSG in the internal phase is prepared using 885 mg of MeDSG according to a liposome preparation method using a reverse phase evaporation method. By adding physiological saline to this gel, dispersing it, and performing a centrifugation operation, excess MeDSG in the aqueous phase outside the liposomes is washed away, and MeDS
The concentration of G is 7. kg/5ffi of the invention product 1 at 11.6
I got d.

Example 2゜DPPC670mg, DOPC440mg, DPP
S 74mg, Cho196o+g and MeDS
765 mg of MeDSG was prepared in the same manner as in Example 1.
Inventive product 2 with a concentration of 1.3 aag/ml for 13.5 d
Obtained.

Example 3゜DPPC670mg, DOPC440mg, DPP
74 mg of S and Cho196o+g were prepared into an aqueous lipid dispersion according to the portic swing method. A physiological saline solution of MeDSG was added to this lipid dispersion, and MeD
Inventive product 3 with an SG concentration of 10I10l1 was 12.5a
I got f.

Example 4 670 mg of DPPC, 440 mong of DOPC, 5511 g of dicetylphosphide (DCP), Chol
96o+g and 853 mg of MeDSG were operated in the same manner as in Example 1 until the concentration of MeDSG was 6.7 mg/ail.
Inventive product 4 of 10.6 was obtained.

Example 5゜ According to the component composition and preparation method of Example 1.

A reversed-phase gel containing MeDSG was prepared. 1 Invention product 5 was obtained. 9 Invention product 5 can be administered orally as it is, but it may also be administered by suspending and dispersing it in water before use. .

Example 6゜The preparation of Example 1 was further supplemented with 40 mg/mi mannitol.
Add so that This solution was freeze-dried and powdered to obtain product 6 of the present invention.

Example 7: Mannitol is further dissolved in the preparation of Example 4 to give 80 rag/d. Spray-dry this liquid to powder 1
Product 7 of the present invention was obtained.The product of the present invention can be administered orally as it is, or can be used as a syrup by dispersing it in water before use. The product of the present invention can also be further processed into dosage forms such as capsules and single tablets.

Example 8 4 QOmg/L light anhydrous silicic acid was added to the preparation of Example 4.
Add and disperse so that it becomes ad. This liquid was spray-dried and powdered to obtain a product 8 of the present invention.

Example 96 Same as Example 4 except that MeDSG in Example 4 was changed to N-(4-(3-aminopropyl)aminobutyl)-2-(7-guanidinohebutanamide)-2-hydroxyethanamide (DSG). In the same manner, 15 relatives of the present invention product 8 having a DSG concentration of 9.0 ig/ml were obtained.

Example 10゜The same procedure as in Example 4 was carried out except that MeDSG in Example 4 was changed to spagarin, and the concentration of spagarin was 8.5 mg/m
12 m of product 9 of the present invention was obtained.

Example 11゜The lipids of Example 1 were added to egg yolk lecithin (Egg PC) 10
80+ag, Chol 96 mg, except that spagarin was replaced with N-[4(3-phenylglycylaminopropyl)aminobutyl II-2-(4-(p-guanidinophenyl)butanamide)-2-hydroxymethylethanamide. The same procedure as in Example 1 was carried out to obtain 12.5 d of product 11 of the present invention having a spagarin concentration of 8.0 mg/ml.

Control example 1. (Control hole 1) Control hole 1 was prepared by dissolving MeDSG in water to a concentration of 10 mg/d.

Figure 1

[Brief explanation of the drawing]

FIG. 1 shows the changes in blood concentration when a product of the present invention and a control hole were orally administered to rats. Fig. 1 Invention product 1 Invention product 2 Invention product 3 Control hole 1

Claims (1)

[Claims] (1) Spagarin preparations characterized by containing spagarin and complex lipids