DE1768836C3 - Process for the production of bis- (betachloräthylamino) -methylrutinen as well as the octagonal bracket on bis- (betachloräthylaminomethyl) square bracket on -rutin and the 6,8- curly bracket on triangular bracket on bis- (betachloräthylaminomethyl) square bracket closed curly bracket to -rutin - Google Patents

Description

in which R is a hydrogen atom or the group

-CH ₇ -N

CH ₂ -

CH ₂ -CH ₂ -Cl

means or from the I-ornnel II

Cl-H ₁ CH ₁ C

\ HO

NH ₂ C! II O-QH _{10 O 4} -O-QH ₁₁ O ₄

Cl-H ₂ CH ₂ C

CI H ₂ CH ₂ C

HO

N-H, C

Cl H ₂ C-H, C

HO O -C ₁₁ H ₁₀ O ₄ -O-QH ₁₀ O ₄

which is characterized in that in a manner known per se for compounds of the general Formula I rutin with bis (// - chloroethylamine) and formaldehyde in a molar ratio of 1: 1: 1 or 1: 2: 2 and for the compound of the formula II in a molar ratio 2: 2: 3 reacts in the presence of an inert solvent or diluent and also relates to the invention the 8-f bis (/ i-chloroethylaminomethyl)] - rutin as well as the o ^ -lDi-rbis-l / i-chlorothylaminomethyl)]} - rutin.

It has been found that the inventive Rutin derivatives of the formula I and II have valuable chemotherapeutic properties. They are cancerostatic effective. A large number of N-Lost connections have already been described, welehe have proven to be more or less effective cancerostatically. Only that was useful Cyclophosphamide (N, N-bis (/) - chloroethyl) -N.O-propylcn-phosphoric acid ester diamide). Here is the cancerotoxic nitrogen mustard group by reaction with phosphoric acid converted into a favorable form of transport, with those responsible for the biological effect Halogen atoms of the nitrogen mustard have been converted into an amide of phosphoric acid. Through this compounds of the cyclophosphamide type are ineffective in vitro against tumor cells. The desired effect only occurs in vivo through enzymatic or hydrolytic cleavage of the phosphamide bond a cancerostatic effect (cf. H. Arnold and H. Klose, "Arzneimittel-Forschung" 10. p. 288 [1960], and 11. p. 159 [1961]). This conversion of the inactive form of the cyclophosphamide into the active form takes place Outside the tumor, preferably in the liver (see R. Rauen, "Arzneimittel-Forschung" 14. Pp. 855 to 859 [1964]). Cyclophosphamide is not cytostatically effective in tissue culture, even then not if the substance is used for 2 hours at 37 C with homogenates of normal and neoplastic Tissues is cultivated. The experimental setup of such a test, which is called an incubation test according to S c h m ä h I and Druckrey is known, is based on the fact that the cancer cells in vitro with the test substance incubated. After washing out the tested connections, the transplantability becomes tested on untreated test animals.

Experiments were carried out using ascites cells of Yoshida sarcoma of the rat. After the cells have been pretreated and incubated with the test substance, they are transplanted with 6 · 10 ⁶ cells / animal. The loss of transplantability after 90 days of the experiment is considered to be the criterion for the effect.

The following table 1 gives the results for the cytotoxic concentration of the tested substances (EL ₅₀ = mean effective concentration):

Table 1

substance

N-mustard (bis- (2-chloroaldehyde) amine).
Cyclophosphamide

I (example I)

II (example 2)

III (example 3)

Yoshida sarcoma in vilro (37 C) IX ₅₀ in μρ ml

4000

15th

21

In Table 2 below, the toxicity data and the therapeutic index indicated.

The compounds in question were chemotherapeutically tested on Yoshida sarcoma in rats.

DL ₅₀ in mg / kg intrapcritoncal.

DC ₅ ,, curative dose which is effective in 50% of the animals.

substance

I (example 1) ..

II (example 2).
IH (example 3)

DL ,,) intraperuoneally of rough

(mg kg I

125
100
140

IX'mi ing (mg kg)

30th
25th
30th

More therapeutic index

IX

16

4th 4th 4. 66

get 25 g of 8- [bis- (, .- chlorolhylaminomethyl) J-rulin from F. 185 to 187 C. The connection sinters to a brownish mass that flares up at 188 to 190 C ".

The crystals can be cleaned by dissolving them in a little acetone and precipitating them with ether or isopropanol will.

Example 2

6,8-; di- [bis - (,; - chloroethylaminomethyl)]! - rutin

The experiments show that i'yclophosphamid has no effectiveness in vitro, but the compounds according to the invention do. These are less effective than nitrogen mustard, but considerably less toxic. This also explains a Different mechanism of action of the compounds according to the invention compared to cyclophosphamide. the Compounds according to the invention apparently attack cancer cells directly in vivo. due the good solubility in water, the stability of the same and the desired intervention against the cancer cell in vivo. In addition, the therapeutic index is very favorable and is only 1 for N-Mustard.

These results and properties are completely unexpected and unforeseeable. The compounds of the invention represent a real advance in that rutin itself is one is a biologically active substance and is a form of transport for the cytotoxic effect of nitrogen mustard particularly suitable.

The new compounds according to the invention are intended to be used as medicaments in human and veterinary medicine Find use.

Example I.
8- [bis (/ - i'-chloroethylaminomethyl)] rutine

22.1 g of rutin (with 3 mol of water) are suspended in about 60 ml of methanol. 4.5 g of bis ( ₍ >'- chloroethyl) amine are stirred into this suspension, and then 3 ml of 37% aqueous formalin solution are added. The mixture is stirred for 10 minutes, during which time slight warming occurs The solution is heated to 60 to 65 ° C. for up to 20 minutes, during which time it dissolves. The color of the solution is reddish yellow and strikingly bright. After everything has been dissolved, the mixture is left to stand for 10 to 12 hours, preferably overnight Clear solution in 100 ml of isopropanol is stirred in portions. Yellowish, matted, clearly recognizable noodles precipitate. After standing for three hours, the isopropanol is decanted, the deposited crystal layer is displayed with 30 to 50 ml of ether and left to stand again for 1 hour, then suctioned off so that the crystals no longer smell of ether. The crystals are then dried at 50 to 50 ° C. Man

35

40

45

22.1 g of rutin (with 3 mol of water) are suspended in 50 ml of methanol, the suspension is 9 g Bis - (, »'- chloroethyl) amine verset / t. and then become 6 ml of 37% formalin solution were added. There is a significant temperature increase of 10 ° C. The mixture is then stirred for 10 minutes and then warmed to 50 ° C. on a water bath the temperature suddenly rises to 70 ° C, dissolving the ruin. You let it cool down and let it go the reaction mixture 12 to 15 hours on its own. Then the clear reddish solution in about 100 ml Poured isopropanol. A bright yellow ul turns out. After standing for 2 hours, it becomes almost colorless supernatant isopropanol solution decanted from the yellow ul. The yellow Ul is gelatinized with 50 ml of isopropanol. slowly making a crunch assumes a crystalline nature. After a few hours, the UI becomes crystalline, you suction off and the substance dries immediately in vacuo at 40 to 50 ° C. About 25 g of 6.8-1 di- [bis - (/; - chloroethylaminomethyl)]! - rutin, which sinters at 85 to 95 C. The substance flares up at 100 C.

Example 3

8-; Di-f6.6'-bis - (/; - chloroethyl-aminomethyl-rutin) j! -
melhan

22.1 g of rutin (with 3 mol of water) are suspended in 50 ml of methanol. One carries into the suspension first 9 g of bis (/; - chloroethyl) amine, then 20 ml of 37% formalin. There is an increase in temperature at 10 C. Now the whole thing is on a dimi water bath heated for another 5 to 10 minutes until complete dissolution and the reaction mixture over Left to itself at night. The reddish brown solution is then poured into 100 ml of isopropanol, a yellow fluffy precipitate separates out, which becomes oily on standing. After standing for 4 hours the isopropanol is decanted and the Ul stirred with acetone, which becomes crystalline. Man repeat the treatment with isopropanol and acetone twice more. use the then received Crystals off and the substance dries quickly in a vacuum, as the crystals strongly attract moisture. About 24 g of 8- | di- [6,6'-bis - (/ i'-chloroethylaminomethyl-rutin)] are obtained ! -methane, which sinters at 100 C and becomes black-brown with effervescence at 156 C.

Claims (1)

Patent claims:
1. Process for the preparation of bis (, - ch! Oräthylamino) -methylrutinen of the general formula 1 HO O
R. HO H ₁ CN in which R is a hydrogen atom or the group CH ₂ -CH ₂ -Cl CH ₁ -CH ₁ -Cl CH ₂ -CH ₁ -Cl -CH ₁ -N means or of the formula II
CI-H ₁ CH ₁ C Cl-HX-H ₁ C CH-CH-Cl HO O
NH ₁ CI II O _{-C 6} H _n AO-C ₁₁ H _{11 O 4} HO HO Cl-H ₁ CH ₁ C NH ₁ C
/ HO O Cl-H ₁ CH ₁ C OH OH O-CH _{10 O 4} -OC ₁₁ H ₁₁ O ₄ characterized in that in a known manner for compounds of general formula 1 rutin with bis (^ - chloroethylamine) and formaldehyde in a molar ratio of 1: 1: 1 or 1: 2: 2 and for the compound of formula M in a molar ratio of 2 : 2: 3 reacted in the presence of an inert solvent or diluent.
2. 8- [bis (/) - chloroethylaminomethyl)] rutin and 6,8-iDi- [bis - (/; ^ iloräthylaminomethyl)]! - rutin. The invention concerns ^; fft a process for the preparation of bis-l / i'-ehloräthylaminol-methylrutin of the general formula I. HO O
RI II O _{-Q 1} H _{111 O 4} -OQ ₁ H ₁₁ O ₄ V-OH HO M ₅ CN CH ₁ -CH ₁ -CI