JPH02157291A - N-formylamino acid conjugate of anthracycline-based antibiotic substance and platinum complex composition thereof - Google Patents

Abstract

NEW MATERIAL:An anthracycline-based N-formylamino acid or N- formyloligopeptide conjugate. USE:A carcinostatic agent. PREPARATION:An anthracycline-based antibiotic substance expressed by formula I (R1 to R4 are H, OH or methoxy; R5 is alkyl, acetyl or hydroxymethylcarbonyl; R6 and R7 are H or methyl), etc., is reacted with an N-formylamino acid or an oligopeptide thereof under neutral to slightly alkaline conditions to afford the objective compound expressed by formula II (R is C-terminal residue of amino acid, oligopeptide), etc.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an N-formyl amino acid or N-formyl oligopeptide complex of an anthracycline antibiotic useful as an anticancer agent and a platinum complex composition thereof.

[Prior art] Rhodomycin is used to treat Streptomyces plubracens (
Streptomyces purpurascens
), more than 30 anthracycline antibiotics have been isolated from the genus Streptomyces. Among them, daunomycin (daunorubicin), adriamycin (doxorubicin), aclacinomycin, etc., which are clinically used as anticancer drugs, have extremely remarkable anticancer effects and occupy an important position in chemotherapy for various tumors.

By the way, it is a clinically difficult point for the above-mentioned anticancer drugs that they also have large side effects. For example, severe cardiotoxicity, hematopoietic organ disorders, hair loss, etc. are observed with adriamycin, and daunomycin has similar tendencies.

[Problem to be solved by the invention]

An object of the present invention is to provide a new substance that reduces the problematic side effects of anthracycline antibiotics and enhances its anticancer effect.

[Means to solve the problem]

As a result of intensive research to solve the above problems, the present inventor has developed an anthracycline antibiotic into a complex with N-formyl amino acid or its oligopeptide, which has a stimulating effect (activating effect) on the human immune system. , and that a highly useful anticancer agent can be obtained by converting the complex into a platinum complex, thereby completing the present invention.

The amino acids and oligopeptides of the present invention differ from conventional anthracycline antibiotics in that they use N-formylated molecules and that they are introduced into anthracycline antibiotic molecules without CONH binding. Essentially different from matter.

In the N-formyl amino acid or N-formyl oligopeptide complex of the anthracycline antibiotic of the invention of item 1, the anthracycline antibiotic used as a raw material is a compound represented by the following structural formula (I). Included.

(In the formula, R to R4 represent a hydrogen atom, a hydroxyl group, or a methoxy group, R3 represents an alkyl group, an acetyl group, or a hydroxymethylcarbonyl group, and R6 and R7 represent a hydrogen atom or a methyl group.) Above- Specific examples of compounds included in maximum (1) include adriamycin, daunomycin, aclacinomycin, and the like.

The N-formyl amino acid or N-formyl oligopeptide complex of the anthracycline antibiotic of the present invention is
In a suitable solvent, under neutral to slightly alkaline conditions, -C compound represented by formula (1) is combined with N-formyl amino acid represented by general formula (II-a) or general formula CII-b),
It is produced by reacting dipeptides, tripeptides, etc. represented by (IT-c) and the like.

General formula CU-a, b, c) [ILa]:]0IIC-NH-CIlR8-COR
9II-b) :0HC-NH-CHRe-CONH-
(jlRe-CORq[11-c:l :0tIC-N
H-CHRe-CONH-CHReCONH-CHR8
-CORqThe amino acids and oligopeptides in the above formula are included as components of normal proteins, except that they are N-formylated, and R8R5,R=
indicates the side chain of an amino acid. Further, R9 represents a hydroxyl group, an alkali salt thereof, an alkyloxy group, a benzyloxy group, an amino group, or the like. -Specifically, the compounds included in the maximum (n-a, b, c) include N-formylmethionine, N-formylmethionylalanine, and N-formylnorleucylleucylphenylalanine, which have a stimulating effect on the human immune system. , N-formylmethionylleucylphenylalanine and its benzyl esters and benzylamides.

As the reaction solvent, water, methanol, ethanol, dimethylformamide, dimethyl sulfoxide, etc. are used alone or in combination, and water is usually preferred.

Compounds represented by general formula (1) and general formulas (IIa, b,
The reaction ratio of the N-formyl amino acid or N-formyl oligopeptide represented by c) is 0.5 to 2.0 mol, preferably 1 mol, of the latter per 1 mol of the former.

Note that the compound of -C formula (1) becomes unstable when the alkalinity of the solvent becomes strong, so care must be taken not to exceed pH 7 or 8 at all times. Reaction temperature is room temperature to 55°C, reaction time is 2 to 2
4 hours is preferred.

The reaction product obtained as described above can be purified using conventional purification techniques such as fractional crystallization including ultracentrifugation that utilizes the difference in solubility in solvents, column chromatography, and the like.

For the formation of a complex between the anthracycline antibiotic produced according to the first invention and N-formyl amino acid or N-formyl oligopeptide, from the measurement of various physicochemical properties and reactivity, mainly general It is thought that there is a bond as shown in .

IC-NH-λ (In II (III), R represents the C-terminal residue of an amino acid or oligopeptide, and R-R represents the same atom or group as in (I)). The related platinum complex composition has the above-mentioned anthracycline antibiotic N
- formyl amino acid or N-formyl oligopeptide complex (main structure is represented by formula (I[[)) in water or a suitable organic solvent, -a chloroplatinate represented by formula (IV) It is produced by reacting.

Mz pc C1n (rv) (In the formula, M represents sodium or potassium, and n represents an integer of 4 or 6) A chloroplatinate represented by the general formula (rV) and a general formula (II
The reaction ratio of the anthracycline antibiotic represented by [) with the N-formyl amino acid or N-formyl oligopeptide complex is 0.5 to 2 mol of the latter to 1 mol of the former.
．． 0 mol, preferably 1 mol.

Note that the -maximum (II[) compound becomes unstable when the alkalinity of the solvent becomes strong, so care must be taken not to exceed pH 7 or 8 at all times. The reaction temperature is room temperature to 55°C, 155°C is 2
~24 hours is preferred.

The platinum complex composition of the N-formyl amino acid or N-formyl oligopeptide complex of anthracycline antibiotics obtained as described above was purified by ultracentrifugation using the difference in solubility in solvents, which is a normal purification method. It can be purified using fractional crystallization methods, column chromatography, etc.

When the N-formyl amino acid or N-formyl oligopeptide complex of the anthracycline antibiotic of the first invention and the platinum complex composition of the second invention are used as anticancer agents, injections are suitable. However, it can also be used as an external preparation or an insert. The injection may be a conventional aqueous injection solution, a powder filler, or a lyophilized preparation, and is prepared according to a conventional method. External preparations and insertion preparations are
It is prepared and used as a composition by blending common pharmaceutical carriers or excipients by a commonly used method.

Examples and test examples of the present invention will be shown below, and will be explained more specifically.

〔Example〕

Next, a method for producing the N-formyl amino acid or N-formyl oligopeptide complex of an anthracycline antibiotic according to the first aspect of the present invention and a method for producing the second platinum complex composition will be described.

Example 1 [Doxorubicin-N-formylmethionylleucylphenylalanine complex] Doxorubicin hydrochloride 58.0μ (0.1mM) was made into an aqueous solution of 10I, and N-formylmethionyl-methionyl-leucyl-cyclochloride was added to this. Slightly alkaline aqueous solution (pl) 7.6-8.0) containing 43.76 mg (0.1 mM) of phenylalanine LM
Add and react at room temperature for 2-4 hours while stirring. Then, the reaction solution is placed in a ventilation dryer and the water is evaporated at 40°C to obtain a red powder.

Pour 20ml of water into this and stir thoroughly.40-45
After removing the insoluble portion with a glass filter, this aqueous solution was applied to an MCI gel column, and the red-orange solution eluted with slightly alkaline water was lyophilized to obtain the target compound 45.
mg was obtained.

Melting point: 163-187'' C1 I Rv: 3 cm-': 3328.1642.
1580.1530.1446°1404, 1284.
1210.1014.986゜Elemental analysis value: Actual value (χ
): C, 50,46; H, 5,98; N, 4.83 Example 2 [Platinum complex composition of doxorubicin-N-formylmethionylalanine complex] Doxorubicin hydrochloride 58.
01mg (0.1mM) was made into an aqueous solution of 10-, and N-formyl-methionyl-methionyl-alanine 24
．． Add 10d of an aqueous solution containing 73mg (0.1mM) and react at room temperature for 6 hours with stirring.Next, the reaction solution was placed in a ventilation dryer at 10°C for several hours to completely evaporate the water, resulting in a red color. Get the powder. After adding slightly acidic hydrochloric acid, 10x ethanol water, and 0 hydroxyl, the mixture was stirred, and the insoluble portion was removed using a glass filter. The filtrate is applied to an MCI gel column, eluted with slightly alkaline water, and the main fraction of the reddish solution is lyophilized.

Pour 20d of water into this and stir thoroughly to 25-30°.
After C), potassium tetrachloroplatinate(n) 41.5
Add 1 mg (0.1 mM) of 10-aqueous solution and react at as low a temperature as possible (5-10°C) with thorough stirring. After 4-6 hours, the precipitate was centrifuged (20,000
rpm, 20m1n.

4°C) and collect. The precipitate was thoroughly washed with hydrochloric acid and slightly acidic water and then with water, and then dried at low temperature to obtain 50 ml of the reddish-orange target compound.

Melting point = 168-181°C0 IR Shini;cn-': 3476.3356,1650
, 1618.1584141B, 1286.1212
．． 1012.984° Elemental analysis value: Actual value (χ); C,
34,16; H, 4,24; N, 3.15 [Test Example] Mouse leukemia cells L 1210 were injected into 6 BDF cells per group.
2 XIO' mice were transplanted intraperitoneally into mice, and the drug was intraperitoneally administered once a day for 5 days from the day after transplantation.
The life extension rate was measured and the results are shown in the table below.

The following formula was used for the life extension rate.

A: Survival days of the drug-treated group B: Survival days of the untreated group AD-FMLP: Doxorubicin-N to formylmethionylleucylphenylalanine complex〇-FMA-P2: Doxorubicin-N-formylmethionylalanine The evaluation is as follows. It was as expected.

or N-formyl oligopeptide complex and its platinum complex composition.

As is clear from the above table, the N-formyl amino acid complex of the anthracycline antibiotic doxorubicin of the invention of item 1 and the platinum complex composition of the invention of item 2 are:
Doxorubicin's published value (ILS 160 ~ 1
80χ, dose 10X5mg/kg/dayXday)
It was observed that the inhibitory effect was significantly superior compared to that of the present invention.

〔Effect of the invention〕

Claims (2)

[Claims] (1) Anthracycline N-formyl amino acid or N-formyl oligopeptide complex produced by reacting an anthracycline antibiotic with N-formyl amino acid or its oligopeptide under neutral or slightly alkaline conditions. (2) An anthracycline-based N-formyl amino acid or N-formyl oligopeptide produced by reacting a neutral to slightly alkaline aqueous solution of the complex according to claim 1 with potassium or sodium tetra or hexachloroplatinate. Complex platinum complex composition