JPH01313425A - Sugar lactam derivative and depressor of metastasis of cancer cell containing the same derivative - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I [R<1>-R<3> is H, WCO- (W is alkyl, aryl, substituted aryl, aralkyl or substituted aralkyl), with a proviso except the case of R<1>-R<3> are equally H]. EXAMPLE:2-0-[1-(p-chlorobenzoyl)-5-methoxy-2-methylindole-3-acetyl]D-g luco-delta- lactam. USE:A depressor of metastasis of cancer cell. Diffusion or transition of cancer cell is very remarkably depressed and cytotoxicity is very small. PREPARATION:D-gluco-delta-lactam compound expressed by formula II and acid halide expressed by the formula WCOX (X is halogen) are subjected to condensation reaction in the presence of base (e.g., triethylamine) at -15 to 80 deg.C for 1 to 24 hour to afford the compound expressed by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a novel glycolactam derivative that significantly inhibits the formation of metastatic foci of cancer cells, and a cancer cell metastasis inhibitor containing the compound.

[Conventional technology]

Conventional anticancer drugs are mainly drugs that kill malignant cells through the cell-killing properties of substances or via the human immune system.

These drugs are still not sufficient for the treatment of cancer. In addition, surgical treatment and radiation therapy are used to treat solid muscles, and the success rate in terms of removing the primary cancer has greatly improved. However, the most important factor influencing the prognosis of cancer patients treated with these treatments is cancer metastasis.

Therefore, it is expected that the effectiveness of these current various therapies will be further enhanced by suppressing the metastasis of cancer cells, but there are few substances whose main action is anti-metastasis, and they are not used clinically. There is nothing.

[Problem to be solved by the invention]

The present inventors previously discovered that an N-substituted-1-deoquinojirimycin derivative significantly inhibits cancer cell metastasis, and filed a patent application for a cancer cell metastasis inhibitor containing this as an active ingredient.
The application was filed as No.-31095.

An object of the present invention is to provide a more powerful cancer cell metastasis inhibitor and a cancer cell metastasis inhibitor containing the same.

[Means to solve the problem]

The present invention represents the formula [wherein Rl, R2t R3 are the same or different hydrogen atoms or IllCD- group (W represents an alkyl group, an aryl group, a substituted aryl group, an aralkyl group, or a substituted aralkyl group]. provided that R', R2, and R3 are both hydrogen atoms] A cancer cell metastasis inhibitor containing a sugar lactam derivative having the formula (1) and a sugar lactam derivative having the formula (1) above.

D-gluco-δ-lactam, which is a raw material for the synthesis of the glycolactam derivative represented by formula (1) of the present invention, is a metabolic product of actinomycetes, nojirimycin (5-amino-5-deokine-D
- Glucobylanose (Japanese Unexamined Patent Publication No. 43-760, Te
trahedron, 23.2125.1968)
It is a substance that can be easily obtained by oxidizing (Meiji Confectionery Research Annual Report, August 80-84, 1973.
-28375 publication).

The sugar lactam derivative represented by formula (1) of the present invention has this D
It is synthesized using -gluco-δ-lactam as a raw material through the following manufacturing steps.

Method A (wherein W has the same meaning as above and X represents a halogen atom) The reaction is first carried out by reacting the carboxylic acid component WCOO)I ( IQ
has the same meaning as above) with an acid halide synthesis reagent such as thionyl chloride, thionyl bromide, or phosphorus oxychloride to obtain an acid halide (WCOX). The reaction temperature is usually 5 to 100°C, and the reaction time is 1 to 24 hours. The acid halide thus obtained is usually used in the next condensation step without being isolated, only by distilling off the reaction solvent and the produced hydrogen halide under reduced pressure.

The obtained acid halide was dissolved in an inert solvent (N, N-dimethylformamide, a halogenated hydrocarbon solvent.

D-gluco-δ-lactam can be prepared as a solution in an inert solvent (N,N-dimethylformamide, N,N-dimethylformamide, etc.) or without solvent.
dioxane, pyridine, halogenated hydrocarbon solvents, etc.)
is added to a solution or suspension in the presence of an inorganic base such as potassium carbonate or sodium hydrogen carbonate or an organic base such as triethylamine or pyridine for condensation. The reaction temperature is usually in the range of -15 to 80°C, and the reaction time is 1.
~24 hours.

Method B The reaction is carried out using the carboxylic acid component WCOOH in an inert solvent such as N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, or pyridine, if necessary, in the presence of an organic base such as triethylamine, pyridine, or diisopropylethylamine. (lν has the same meaning as above) and a dehydration condensation agent such as 1,3-dicyclohexylcarbodiimide, diethyl cyanophosphonate 1
This is carried out by reacting with diphenylphosphoryl azide, p-toluenesulfonyl chloride, triphenylphosphine-diethyl azodilypoxylate, 1,1°-carbonyldiimidazole, and the like. The reaction temperature is usually 5 to 100°C, and the reaction time is 1 to 100°C.
It is 48 hours.

The compound of formula (1) obtained as described above is collected from the reaction mixture by a conventional method. For example, it is isolated as a pure product by an extraction operation using an organic solvent or water, a precipitation method, a crystallization method, or various chromatography methods.

Examples of the compounds of the present invention include the sugar lactam derivatives shown below. However, it is not limited to these.

2-0-[1-(p-chlorobenzoyl)-5-methoxy-2-methylindole-3-acetyl]-D-gluco-δ-lactam, 6-0-[1-(1]-chlorobenzoyl) -5-methoxy-2-methylindole-3-
acetyl]-D-kururo δ-lactam, 2.6-di0-[1-(11-chlorobenzoyl)-5-methoxy-2-methylindole-3-acetyl]-D-gluco-δ-lactam, 2 -0-[3-benzoylhydratroboyl]-D-gluco-δ-lactam, 6-0-[3
-benzoylhydratroboyl]-D-gluco-δ-lactam, 2,6-di0-[3-benzoylhydratroboyl]-D-gluco-δ-lactam, 2-'0-[6
-methoxy-α-methyl-2-naphthaleneacetyl]-
raw gluco-δ-lactam, 6-0-[6-methoxy-
α-Methyl-2-naphthaleneacetyl]-D-gluco-
δ-lactam, 2,6-di0-[6-medoxy α
-Methyl-2-naphthaleneacetyl]-〇-gluco-δ
-lactam, 6-0-[N-(2,3-xylyl)anthraniloyl]-D-gluco-δ-lactam, 6-0-
(acetylsalicyloyl)-D-gluco-δ-lactam, 2,6-di0-(acetylsalicyloyl)-D
-Gluco-δ-lactam, 2,3.6-)li-0-(acetylsalicyloyl)-D-gluco-δ-lactam, 6
-0-diphenylacetyl-〇-gluco-δ-lactam, 6-0-) diphenylacetyl-〇-gluco-δ
-Lactam.

Next, an example of producing the sugar lactam derivative of the present invention will be given.

Production Example 1 6-0-[6-methoxy-α-methyl-2-naphthaleneacetyl]-D-gluco-δ-lactam D-glucose
δ-lactam 5.31 g (30 mmol), 6-methoxy-α-methyl-2-naphthaleneacetic acid 6.9 g (3
0 mmol) was dissolved in 50 cc of DMF, then 7 cc of triethylamine and 6.7 cc of diethyl cyanophosphonate were added and stirred overnight at room temperature. After removing the insoluble portion by filtration, the filtrate was concentrated to obtain 5 g of a residue. The residue was dissolved in ethyl acetate and water, and the ethyl acetate layer was separated and concentrated to obtain 3.8 g of oil. This was separated by silica gel chromatography (chloroform-methanol 10:1)
The title compound [silica gel thin layer chromatography (
Chloroform-methanol 4:1) Rf 0146:]
1.6g was obtained.

Production Example 2 2-0-[6-methoxy-α-methyl-2-naphthaleneacetyl]-D-gluco-δ-lactam and 2.6-di〇=[6-methoxy-α-methyl-2-naphthaleneacetyl ] -D-gluco-δ-lactam 6-methoxy-α-methyl-2-naphthaleneacetic acid 11,
5 g was dissolved in 160 cc of methylene chloride and dimethylformamide Q, lcc, and 7.3 cc of thionyl chloride was dissolved.
was added and stirred at room temperature for 5 hours. After distilling off the solvent, toluene was added and the mixture was further concentrated to dryness to obtain an acid chloride.

Dissolve 5.85 g of raw gluco-δ-lactam in 100 cc of dimethylformamide, and add 12rd pyridine! The mixture was further cooled to 5° C., and the acid chloride of 6-methoxy-α-methyl-2-naphthaleneacetic acid was added thereto, and the mixture was stirred overnight at room temperature. After removing the insoluble portion by filtration, the solvent was distilled off, toluene was added, and the mixture was further concentrated to dryness to obtain a residue. Ethyl acetate and water were added, and the insoluble portion was first collected by filtration. Methanol 10:1) Rfo, 26] 1.8 g
I got it.

Then, the ethyl acetate layer was separated and concentrated to reduce the amount of ethyl acetate to 1
50cc, filter the insoluble part, and filter out the 2,6-di0-[6
-Medoxy α-methyl-2-naphthaleneacetyl]
-〇-Gluco-δ-lactam [Silica gel thin layer chromatography (chloroform-methanol 10:1) Rf
5.86 g of O,48] was obtained.

Production Example 3 6-0-[1-(p-chlorobenzoyl)-5-methoxy-2-methyl-3-indoleacetyl]-D-gluco-δ-lactam and 2.6-di0-[1-( 5.37 g of p-chlorobenzoyl)-5-methoxy-2-methyl-3-indoleacetyl]-D-gluco-δ-lactam indomethacin was dissolved in 50 cc of 1,2-dichloroethane and 0.1 cc of dimethylformamide, and thionyl chloride was dissolved. 2.25 cc was added and stirred at 60°C for 1.5 hours.

After concentration, toluene was added and further concentrated to obtain the acid chloride form of indomethacin.

Dissolve 2 g of D-gluco-δ-lactam in a mixture of 50 cc of dimethylformamide, 1 liter of pyridine, and 7 cc.
Cooled to ℃. Next, a solution of 20 cc of dimethylformamide of the acid chloride form of indomethacin was added, and the mixture was stirred at room temperature overnight. The solvent was distilled off and separated directly by silica gel column chromatography (chloroform-methanol 20%
:1) and 0.7 g of 6-0-[1-(p-chlorobenzoyl)-5-methquin-2-methyl-3-indoleacetyl]-D-gluco-δ-lactam and 2-0
-[1-(p-chlorobenzoyl)-5-methoxy-2
-Methyl-3-indoleacetyl]-〇-gluco-δ
-Lactam [Silica gel thin layer chromatography (chloroform-methanol 1 [1:1) Rf O, 33]
0.7g.

and 2,6-di0-[1-(p-chlorobenzoyl)-5=methoxy-2-methyl-3-indoleacetyl]-D-gluco-δ-lactam [silica gel thin layer chromatography (chloroform-methanol 10 :1)
tuo, 50] 2.16 g was obtained.

Production Example 4 2.6-D-[3-benzoylhydratroboyl]
-D-gluco-δ-lactam 5.08 g of ketoprofen, 50 cc of 1,2-dichloroethane and 0.0 cc of dimethylformamide. It was dissolved in a mixed solution of ICC, 2.95 cc of thionyl chloride was added, and the mixture was stirred at 60°C for 2 hours. The solvent was distilled off, toluene was added, and the toluene was further distilled off to obtain an acid chloride of ketoprofen.

3.54 g of D-gluco-δ-lactam was dissolved in a mixed solution of 35 cc of dimethylformamide and 4 cc of pyridine and cooled to 5°C. A 20 cc solution of the acid chloride of ketoprofen in dimethylformamide was added, and the mixture was stirred at room temperature overnight. The solvent was distilled off, and 4 Qcc of water, 4 Qcc of methanol, and 150 CC of ethyl ether were added to the residue to make a two-layer solution, and the ether layer was separated. Further, the ether layer was washed with aqueous sodium bicarbonate, the solvent was distilled off, and then separated by silica gel column chromatography (chloroform-methanol 40:1) to obtain the title compound [silica gel thin-layer chromatography (chloroform-methanol 10:1).
1.98 g of Rf O,44] was obtained.

Production Example 5 6-0-[N-(2,3-xylyl)anthraniloylco-D-gluco-δ-lactam D-kururo 5.31 g of δ-lactam and 7.23 g of mefenamic acid were added to 60 cc of dimethylformamide. After dissolving, 6.3 cc of triethylamine and then 6.6 cc of diethylcyanophosphonate were added, and the mixture was stirred at room temperature overnight.

After evaporating the solvent, ethyl ether was added and the mixture was stirred at room temperature, and the insoluble portion was filtered. A small amount of methanol was added to the solid matter collected by filtration to form a solution, and then ethyl ether and isopropyl ether were added and stirred, and the precipitate was collected by filtration. The obtained precipitate was further made into a methanol solution and reprecipitated by adding isopropyl ether to obtain the title compound [silica gel thin layer chromatography (chloroform-methanol 4:1) Rf O
, 51) 1.42 g was obtained.

Production Example 6 6-0- (acetylsalicyloyl)-D-gluco-δ
-lactam, 2,6-di-0-(acetylsalicyloyl)-〇-gullo δ-lactam and 2,3.6-di-0-(acetylsalicyloyl)-〇-gullo δ-
3.52 g of lactamlogluco-δ-lactam was suspended in a mixture of 50ml of pyridine and 11N of N-dimethylformamide, and 8.0g of 0-acetylsalicyloyl chloride and 50rn of N,N-dimethylformamide were suspended under water cooling.
! The solution was added dropwise.

After stirring at 0 to 5°C for 1 hour, the mixture was returned to room temperature, stirred overnight, and then the insoluble portion was filtered off and concentrated to dryness. The residue was subjected to silica gel column chromatography (chloroform-methanol 10:1).
) and purified with 6-0- (acetylsalicyloyl)-
D-gluco-δ-lactam [silica gel thin layer chromatography (chloroform-methanol 10:1) Rf
O,11) 0.3 g, 2.6-di0-(acetylsalicyloyl)-D-gluco-δ-lactam [Silica gel thin layer chromatography (chloroform-methanol 10:1) Rf O,41) 3 ．． Og, 2,
3.6-)G0-(acetylsalicyloyl)-〇-
Gullo δ-lactam [Silica gel thin layer chromatography (chloroform-methanol 10:1) Rf O
, 53) 0.4 g were obtained, respectively.

The cancer cell metastasis inhibitor of the present invention is clinically administered as an oral or parenteral preparation containing the above-mentioned 0-acyl-D-gululo δ-lactam via veins, arteries, skin, subcutaneous, intradermal, rectal and intramuscular routes. Or administered orally. Moreover, stronger effects can be expected by administering directly to the tumor. Although the dosage varies depending on the dosage form, dosage form, age, weight, and pathological condition of the patient, it is generally administered at 100 to 3000 mg once or several times a day.

Parenteral preparations include sterile aqueous or non-aqueous solutions or emulsions. Examples of the base for non-aqueous solutions or emulsions include propylene glycol, polyethylene glycol, glycerin, olive oil, corn oil, and ethyl oleate. Oral preparations include capsules, tablets, granules, fine granules, powders, and the like. These preparations contain starch, lactose, mannitol, ethylcellulose, sodium carboxydimethylcellulose, etc. as excipients, and magnesium stearate or calcium stearate is added as a lubricant. As the binder, gelatin, gum arabic, cellulose ester, polyvinylpyrrolidone, etc. are used.

In addition, 2,6-G0- which is one of the active ingredients of the present invention
The acute toxicity (LDso value) of (acetylsalicyloyl)-D-gluco-δ-lactam in mice is 0.5 g/kg or more after 1p administration.

Next, formulation examples and effects of the present invention will be explained.

[Example] Example 1 6-0-[1-(p-chlorobenzoyl)-5-methoxy-2-methylindole-3-acetyl]-D-glu'ro δ-lactam 50m
g Lactose 130 mg Potato starch 70 mg Polyvinylpyrrolidone 1 (1mg Magnesium stearate 2.5mg Mix lactose and potato starch, add a 20% ethanol solution of polyvinylpyrrolidone to the mixture, moisten it evenly, and pass through a 1-mesh sieve. , dried at 45° C. and passed through again. The granules thus obtained were mixed with magnesium stearate and formed into tablets.

Example 2 6-0-[N-(2,3-xylyl)anthraniloyl]-D-gluco-δ-lactam is ground with a pin mill to particles of approximately 10μ, and 500 mg thereof is filled into an injection vial. . Separately Tween 80
5 mg and 10 mg of diurnal purified gelatin are dissolved in distilled water for injection 5 to prepare a solution. When using, add this solution 5- to the fi-0-[~(2,3-xylyl)anthraniloyl]-D-gluco-δ-lactam filled in the vial and shake well to make a suspension. Administer an appropriate amount intramuscularly.

Efficacy test method
(Fiddler) Method in Can
cer Re5ear-ch, Hen, 399-439.1
978), the B16 highly metastatic strain was selected and used.

The metastasis suppressive effect was evaluated using Kijimasuda (Kijima-3).
uda) et al.'s method (Cancer Research
h, 46,858-862゜1986).

First, the 816 highly metastatic strain was planted in Dulbecco's medium supplemented with fetal bovine serum, and 0-acyl-D-gluco-
Add delta-lactam and 5% CD for 2-4 days.

The cells were cultured at 37°C in the presence of , and the proliferated cells were detached from the culture vessel using a trypsin and BDTA solution. The cells were suspended as living cells in a balanced salt solution at 1×10 6 cells per cell.

0.1 d of this suspension was transplanted from the tail vein of a mouse, and
After being reared for one day, the lungs were removed through laparotomy, and the nodules of the 816 highly metastatic strain formed on and inside the lungs were counted and compared with a control that was not treated with the drug.

Test Example 1 Cytotoxicity of 2,6-di-o-[1-(p-chlorobenzoyl)-5-methoxy-2-methylindole-3-acetyl]-D-gluco-δ-lactam B16
10% of highly metastatic strains and mouse-derived tumor cells, 929 strains
After culturing in Dulbecco's medium supplemented with tallow baby serum at 37°C in the presence of 5% C02, the cells were detached from the culture vessel with a trypsin and EDTA solution.
Xl0' cells, strain 929, were suspended in the above medium to become IXIO3 cells.

1504 of this suspension was added to 2,6-di0-[1-(p-
chlorobenzoyl)-5-methquine-2-methylindole-3-acetyl]-D-kururo δ-lactam or adriamycin (control) solution (50μ) was added and mixed. For 816 highly metastatic strains, IO
The 29 strains were cultured for 4 days, and their viability was determined under an inverted microscope to determine cytotoxicity. The results were as shown in Table 1.

(The following is the margin of this page) Table 1 Test Example 2 2, 6-C[]-(Acetylsalineloyl)-D-
Anti-metastatic effect of gluco-δ-lactam A B16 highly metastatic strain was planted in Dulbecco's medium supplemented with 10% tallow serum, and 2.6-di0-(acetylsalicyloyl)
-〇-1ml of Gullo δ-lactam! 10g per
In addition, the cells were cultured at 37° C. for 3 days in the presence of 5% CO2. Peel the cells from the culture container in the same manner as in Test Example 1, and
Suspend the living cells in balanced salt solution to 1 x 106 cells per liter, and add 0.1 d of the live cells to BDF, mouse (8 weeks old,
It was administered into the tail vein of a male (male) and transplanted. After 14 days of rearing and observation, the lungs were removed through abdominal surgery, and B formed on the surface and inside of the lungs was observed.
Nodules of 16 highly metastatic strains were counted. The results are shown in Table 2.

Table 2 From the above test results, 2,6-
G0-(acetylsalicyloyl)-〇-gullo δ
-It can be seen that lactam strongly inhibits metastasis to the lungs.

〔Effect of the invention〕

The cancer cell metastasis inhibitor of the present invention is a drug that very significantly suppresses the spread and metastasis of cancer cells and has very little cytotoxicity. Therefore, when administered simultaneously with existing anticancer drugs,
It is an extremely useful drug that can significantly enhance anticancer effects when used during surgical therapy or radiotherapy.

Claims (1)

[Claims] 1. Formula ▲ Numerical formula, chemical formula, table, etc. ▼ [In the formula, R^1, R^2, R^3 are the same or different hydrogen atoms or WCO- group (W is an alkyl group, aryl group, substituted aryl group, aralkyl group, or substituted aralkyl group). However, this excludes the case where R^1, R^2, and R^3 are all hydrogen atoms.] A sugar lactam derivative having the following. 2. Formulas ▲ Numerical formulas, chemical formulas, tables, etc. , represents an aralkyl group or a substituted aralkyl group). provided that R^1, R^2, and R^3 are all hydrogen atoms] A cancer cell metastasis inhibitor characterized by containing a glycolactam derivative having the following.