DE1768836B2 - 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

i 768

in the R a WassenrtofTuioro or the group

• —CHj-N

means or of the formula II

CH, -CH, -Cl

CH, -CH, -Ci

Cl-H ₂ CH ₂ C

\ HO

M — tr c I Il r \ Λυ η

CH ₁ CH ₁ C YJ ^^ VOH

HO I

OH

CH ₁

(II)

Cl-HX-H ₁ C

N-HX I H \ OH

HO O \
Cl-H ₂ C-HX 0-Q _{1 111} O H ₄ -O-C ₆ H ₁₀ O ₄

which is characterized in that in a manner known per se for compounds of the general Formula I rutin with bis-ijf-chloroethylamine) and formaldehyde in a molar ratio of 1: 1: 1 or 1: 2: 2 and for the compound of Fo: .tiel II in a molar ratio 2: 2: 3 reacts in the presence of an inert solvent or diluent and also relates to the invention is 8- [bis (/ * - chloroethylaminomethyl)] rutin and 6,8-iDi- [bis - (/ i-chloroethylaminomethyl)]} - rutin.

It has been found that the rutin derivatives of the formula I and II according to the invention are valuable chemotherapeutic Possess properties. They are cancerostatic. There are already a large number of nitrogen mustards has been described, which are more or less cancerostatic have proven effective. Only the cyclophosphamide (N, N-bis - (// - chloroethyl) -N.O-propylene-phosphoric acid ester diamide) was usable. This is where the N-Mustard group, which is toxic to cancer, occurs through 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 occurs only 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 treated with normal and neoplastic homogenates for 2 hours at 37 ° C Tissues is cultivated. The experimental setup of such a test, which is an incubation test according to S c h m ä h 1 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 animal cells. The loss of transplantability after 90 days of testing is used as a 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-chloroethyl) amine)
Cyclophosphamide

I (example I)

II (example 2)

III (example 3)

Yoshida sarcoma in vitro (37 C) EL50 in μg ml

4000

15th

21

The following table 2 shows 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 ₅₀ curative dose that is effective in 50% of the animals.

substance

! (Example I) ..
I! (Example 2).
IH (example! 3)

Pl-S ₀ jniraperltoneal
on rats

(mg / leg)

125
100
140

PC's so ine
Irng / kg)

30th
25th
30th

TherapcH-tiscncr index

PU
PC ₅₀

4. 16 4 4. 66

The tests show that cyclophosphamide has no activity in vitro, but the compounds according to the invention do. These are actually less effective than nitrogen mustards, but considerably less toxic. This also explains another mechanism of action of the compounds according to the invention in relation to cyclophosphamide. The compounds according to the invention apparently attack the cancer cell directly in vivo, due to 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 n ^r r 1 for nitrogen mustards.

These results and properties are completely unexpected and unforeseeable. The compounds according to the invention tax! a real progress insofar as rutin is itself a biologically active substance and is particularly suitable as a form of transport for the cytotoxic effects of nitrogen mustard well suited.

The new compounds according to the invention should are used as remedies in human and veterinary medicine.

Example I.

H-LBis-l / i-chloroilylaminomicihylJJ-riUin

22.1 g of rutin (with 3 mol of water) are suspended in about 60 ml of methanol. 4.5 g of bis (i-chloroethyl) amine are stirred into this suspension, and then 3 ml of 37% strength aqueous formalin solution are added. The mixture is stirred for 10 minutes. with slight heating occurring. The mixture is then heated to 60 to 65 ° C. on the water bath for 15 to 20 minutes, during which time it dissolves. The color of the solution is reddish yellow and strikingly bright. After everything is resolved. the reaction mixture is 10 to 12 hours, expediently overnight. ditched. The clear solution is then stirred into 100 ml of isopropanol in portions. Yellowish, matted, clearly recognizable noodles fall out. After standing for three hours, the isopropanol is decanted, the deposited crystal layer is made into a paste 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. 25 g of B-tBis-i / i-chloroethylaminomethylrmin from V. J 85 to 187 ° C. are obtained.

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

Example 2

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

22.1 g of rutin (with 3 mol of water) are suspended in 50 ml of methanol, the suspension is 9 g Bis - (/ * - chloroethyl) -atnin is added, and then

Every 6 rnl 37% formaldehyde solution Tiigegihcn Fs occurs a significant temperature increase of 10 C. The mixture is then stirred for 10 minutes and heated then on the water bath to 50 ° C, the temperature suddenly dissolving the rutin

rises to 70 C. It is now allowed to cool and the reaction mixture is left to look for 12 to 15 hours. Then the clear reddish solution is in about 100 ml Isopropanol poured.:. A bright yellow oil falls the end. After leaching for 2 hours, the almost colorless supernatant isopropanol solution is decanted from the yellow oil. The yellow oil is diluted with 50 ml of isopropanol. wöbe: it's slowly crunching assumes a crystalline nature. After a few hours, the oil becomes crystalline, it is suctioned off and the substance dries immediately in vacuo at 40 to 50 C. This gives eP-va 25 g of 6,8-! di- [bis - (,; - chloroethylaminomethyl)] ! -rutin, which sinters at 85 to 95 C. The substance flares up at 100 C.

B e i s ρ i e 1 3

S-lDi-f, 6.6'-bis - (,; - chloroethyl-aminomethyl-rutinl]; - methane

22.1 u rutin; with 3 mol water) are dissolved in 50 ml Suspended methanol. First 9 g of bis-lj -'-chloroethyl / amine are added to the suspension. then 20 ml of formalin 37 "above. The temperature increases by 10 C. Now the whole thing is on the 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 decanured and the oil is stirred with acetone, this *; becomes crystalline. Man repeat the treatment with isopropanol and acetone twice more. sucks 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 - (/; - chloroethylaniinomethyl-rutin) j} -methane are obtained, which sinters at 100 C and becomes black-brown from 156 C with effervescence.

Claims (1)

i 768 836 ι * Patent claims:
I. Process for the preparation of bis (// - chloro-thylHmiRo) -methylr «tmen of the general formula HO O RT Ii HO KX-N in which R is a hydrogen atom or the group -CH ₁ -N CH ₂ -CH ₂ -Cl CH ₂ -CH ₂ -Cl CH-CH-Cl means or of the formula II
Cl-HX-H, C \ HO O N-HX II! 0-C ₀ H ₁₀ Ov-OC ₆ H ₁₁ O ₄ Cl-H ₂ C-HX CH ₇ C-HX Cl-HX-HX HO HO N-HX OH OH CH, OH Ί
OH HO O O-CH _{10 O 4} -OC ₆ H ₁₁ O ₄ 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 the compound of the form! II in a molar ratio of 2: 2: 3 in the presence of an inert solvent or diluent.
2. S-fBis-lp'-chloroethylaminomethylO-rutin and oS-iDi-Cbis-l / i-chloroäthylarninometlivirii-rutin. The invention relates to a process for the preparation of bis- (, <- chlorathylamino) -methv irulin of the general Formula I. HO O
R i Ü 0-CH ₁₀ O ₄ -O-CH ₁₁ O ₄ HO T- OH OH
CH ₂ -CH ₂ -Cl HX-N CH, -CH, -Cl