SU1261253A1 - Oxoammonium derivatives of nitrosourea displaying antitumor activity, and method of producing same - Google Patents

Abstract

1. Oxoammonium derivatives of nitrosoureas of the general formula 'R' ^ CH ^) ^ m ^ C

Description

H, R. to H, H, R - NO, - H, R 3 HO, K, R - NO, CH ,, R / NO, R - sn ;. R: CHjCH, tCl, NO (Villa) and R H, R R CHjCH Cl, n 2, H, NO (IXa) and R H, R NO, R R R R

NZ, SNZ

 ,

HjC CHj

exhibiting antitumor activity.

2. A process for the preparation of oxoammonium nitrosourea derivatives (I-IV, VX, VI11-X), consisting in the tone that the amines R (CH.), - NHRHR - 2.2, 6.6-tetramethyl -1-oxyl-4- pyridip or NO, R NO, .RR

to

JV

O9

4-to arboxy-2,2,6,6-tetramethyl-1-oxyl-4-piperidnl) or R. isocyanates (NCO, respectively, isocyanates with RNCO isocyanates or amines R KHj at equimolar ratio of reactants, temperature from -10 to 30 C to nitroxyurea, R (C1) NR C (0) NI1R, the last oxide N0 (I); O, X3, n O, x C1- (II): C1, n "0.1x NO; (III); IHjCHjCl, n o, X-- CI- (IV); CH, CH, C1, n O, iX-C10 (V); CH, CHjCl, n 0, - (VI); .Cl, n 0, yC ClOf (VII); mixture of isomers NO, RH (VIII6); X-Cl; mixture of isomers Ci, RH (1X6); X-NO; (X); SI CH Cl, n 0,

ly1K5t equivalent to KonKqeeicBOK dn ™ zhor in an inert solvent with a temperature range from 0 to

.-50, C, .os are waiting for an excess of dry ef11ra oxoam1

I (SNRC NRC (0) tffiR Cl, separate them from the mother liquor of nitroeurucleos in | for 0.5-.3 h equimole rsn: the amount of tetrasyxidiazot or ibytym nitrosylchloride in an inert solvent at a temperature of from O to -OC and after precipitation) Existence of dry zfra is equivalent to compounds (1, III-X),

3. The method according to uhochega oxoachovenus derivatives of nitrosourea, VII), which consists in the production of syngeneic aqueous solutions of proivvodnyh (III, IV) or VI) by treating SJT with an equivalent amount of yenograged perchloric acid and making the precipitated compounds (V, VII) ,

The invention relates to new derivatives of nitrosourea, which can be used in medicine,

The claimed oxoammonium functional derivatives of nitrosourea common fork (0) TDe R 2.25b, b tatrametvl-1-oxopine ™ ridinio 4-sh1 or 4-carboxy-2 2,6S6-tegram 7-sh-1-dsopnpernnio-4-il9. R H or CHg bw KOs N0 or H, lit in CH3 or -chloroethyl, p X NQj or CtOj, or Cf fcioryT are used as anti-tumor drugs. Along with high solubility of water to water, 5 which is a valuable property for medicinal paratytes, oxoammonium nitrosourea derivatives in vivo are easily reduced to paramagnetic nitroxylphenol production of nitrosomeric acid. The latter property of the compounds under study allows their headlights the macologic behavior in the organism using the electron paramagnetic resonance (EPR) method.

The aim of the invention is to obtain new water-soluble Benteci-E with increased activity and reduced toxicity, which would allow 5 days undesirable side effect of the drug, to increase the effectiveness of suppressing malignant tumors in the body. 1SHZM ;, meaning ()) to measure the range of action and control the behavior of the drug in the body

2

Example 1. Preparation of c6-J6 ° -gramethyl-4- (3-matal-reuido) yiperidin-1-oxyl (XVa), 2,2,6,6-teramethyl-4- 3 (2 klorethyl) y |) eido peipo-

p dynoxyl (XV6) :, 2j Zgb.b-tetry g atnp-4 -, {1-Methyl-3- (2-clasher7) urea, ojpiperidine-cx (XVv) 4. 2.2.6, 6 tegramethyl-4- 3- (2-ortzyl) ureadogaty} -pkperi; qing-1 oxyl (XVr),

2.29b, 6-tetramethyl-4- 2-GZ- (2 ™ ylor. Etsh) ureids} etgsh ggiperndik-oxshga HUD) - and 4 Kap6oKCfj-2,2j6t6-TeTp0.se til-4-GZ {2-ztoretSh1 ) urekdo1 pkperi-

dyn-1-oxyla (XVe).

/ -.

Method A, To a solution of .10 mmol of one of the amino radicals (X1a-g) s 5 ml of ether at a temperature of from 0 to 30 s and stirring, add a solution 1 (, 06 g (10 mmol) of isocycate (XII) in 5 MP ether , The reaction mixture is stirred without cooling, the crystalline urea is separated and parecrystallized, and the oily is purified on a column of silica gel, elgate goroform-acetone-methanol. TO: 1; t (see Table 1) -Yyuchev5, (XVe), ipodaty and acetoiitrile with the addition of 5 mol, A

,

Method B. To a solution of fO miol nzocyanate (XIII) or (XBb) in 5 ether with stirring, add 10 ppm of 1 M solution (XlVa) or. (XIV6) on the air. After 0.5-3 hours, the formed ureas are separated and purified as indicated in table. 1, 3 Outputs, melting points, and data for an eunent analysis for urea (XVa-e) are listed in Tab, and their spectral properties are given in Table. 2. Example 2. Received, 6, -tetramethyl-4- | C 3-methylurendo) -1 oxopiperidinium chloride (XVIa), 282,6,6-tetrametesh1-4- 3- (2-TCHOR9TYu ureido -1-oxopersidine chloride XVI6) „2,2,6,6, -tetramethyl-4-.1-m TSH1-3- (27chloroethyl) urevdo-1-oxopyridine chloride; (Х71c)% 2,2,6i, 6 -tetramethyl-4 - {3- (2-chlo1re1Il) ureid methyl T-T-oxopiperidinium chloride (XVIr), 2,2, b, 6-7 tetramethyl 4-C2-13 - (2 x Lorethyl) ureido ethyl - 1-oxopyridine. Chlorine (HUTD ) and A-car6o, 6.6 tetramethyl-4-t3- (2-hlop ethyl) ypeidol-1-oxopnepne chloride (XVIe). To a solution of 10 mmol of the corresponding nitroxylurea (KUa-e) in 5 MP chloroform in a porous bottom reactor and cooling jacket when purging from the bottom with dry argon And at a temperature from 0 to, a solution of 5 MP in 5 MP CCtj is added dropwise. The reaction mixture at a temperature below 0 ° C is stirred for 15–120 min with a weak argon current, —the crystalline chloride precipitated by ethereal of crystalline chloride from the solution is washed, washed with ether, and dried in a vacuum. Chlorides (XVIa-e) are low-stable, at room temperature, crystalline substances of yellow or brick red. colors. For (). Found,%: C A6.5; H 7.12; N 13.7 C ,, N305 Calculated,%: C 46.16} H 7, A2 ,; N 13, (6; 11 312.2А. For (XVIe) it was found,%:,; H 6.40-, N12.05.% H .O, Calculated,%: С A3.83; H 6.51; N, 11.80; ti 356.23. The yields, melting points, and data of IR spectra (XVIa-e) are given in Table 3.., Example 3. Obtaining the claimed compounds: nitrate (1) and chloride, (Ii) 2,2,6, b-tegramethyl-4- (3-methyl-3-nitrosoureo) -1-oxopiperdipipi, nitrate (y) ichloride (IV) 2,2,6,6-tetramethyl-A- 353 .4 nitroso-3 (2-chloroethyl) ureido -1-oxopiperidine I, 2,2,6,6-tetramethyl-4-1-netil-3-nitroso-3- (2-hporethyl) ureido -1-oxopiperidinium nitrate (VI 2,2,6,6-tetramethyl-4- 3-nitroso-3- (2-chloroethyl) ureidomethyl1-1-oxopiperidinium nitrate (Villa) mixed with isomeric 2,2,6,6-tetra-methyl-4 1-nitroso-3- (2-chloroethyl) ureidomethyl1-1-oxopiperidinium nitrate (VIII6) 2,2,6,6-tetramethyl-A- 2 -13-nitroso-3- (2-chlorostil) ureido-ethyl -1-oxopiperidium chloride (1Xa) mixed with isomeric 2,2,6,6-tetramethyl-4- (2- 1-nitroso-3- (2-chloroethyl ) ureido ethyl -1-oxopiperidinium chloride (1X6) H 4-kggrboxy-2, 2,6,6-tetramethyl-A-13-nitroso-3- (2-chloroethyl) ureido -1-oxopiperidinium nitrate (X). 10 mmol of the corresponding chlorides (XVIa-e) (see Ex. 2) immediately after preparation are suspended in 5 ml of chloroform and at a temperature from 0 to TSO C and stirring with argon to the suspension is added dropwise a solution of 10 mmol of P in 7 cpm; ororoform for the preparation of nitrates (1, 111, Vt, V111, X) a solution of 15 mmol NOC1 in 10 ml of chloroform to obtain the chlorides (11, IVi IX) of oxoammonium nitrourea. The cooling bath is then allowed to warm to room temperature. After 0.5-3 hours, the products or precipitated with ether are removed from the reaction mixture, filtered, washed with ether and sugate in a vacuum of 1 Pa. According to the method given in (1), it was found that the derivatives (1-1V, VI, X) are formed - Clues in the form of 3-nitroso 13oHeroz, and (VI11, IX) are mixtures of 3- and 1- nitroso isomers; the ratio of isomers of the mixtures is: Villa (VniG 3: U1Xa) 1X6 1: 1. The yields, melting points, solvent for recrystallization, and data on the elementytome for the analysis of nitrates and oxides of ammonium ammonium hydroxylureas are given in Table. m., and with spectral properties in table. 5. Example A: Preparation of 2,2, 5b-tetramethyl-4- 3-1 nitroso-3- (2-coretm) ureido-β-oiiperidio di perchlorate (V) and 2,2,6,6 -tetraetyl-4- 1-methyl-3-nitroso-3- (2-jpiopsTKn) ureado --1 goxashyo “9th shsorate (yU)” To pacTBbpy 10 shshp 1sh “grvda (fV) or Nitrate (VI) in micron m8Ly oh me A drop of t, 4 from 7: 2Z-Hoft IWfOj is added dropwise to the body. Fine white perchlorate crystals of nutfshit are heated with cold & ether and dried in a vacuum at t over aV -; ---- -:. -. : ,, Conclusions, temperatures p evuy is the data of elemental aialdase (tVv VII are given in tab. -4j and the spectrum spectrum is higher in table. 5. See the fuzzy details of the data that jumped to 4 x e0dggey (-X) and 6Lyi) aiIe analogue of TsGYM, | fivedesh a table 6, Yojiziz "tto connections - (t-IV, yf-, Vttf, IX) obya arg high dissolution; th water and were investigated in the form of waters ah pac-TBopoSj, the rest - eleven; K) I. TSGSh, c. in the form of emul1S in 0%. An aqueous solution of twin 80a. / The toxicity of the compounds described is determined by a single-time test on whites. Without using the Wörns method. As a hawk, there was a dose of gosh, a dose of 50 shots taken in opsh- (). ; The test results showed that the price at Jie Oxoammonievka prokznynye (1-K) t, 5-5 times less. tdksichy than TsGNM,: The determination of the protomoolous AK TRSHpcTH was performed on tumors - - nepeBHaaeMJJJx intraperitoneally leukemia x La L t2tO and Р-388 and pereoed s moe leukemia ascetic Ehrlich cancer (ERE); and kpucoli KPIMC - transplantable subcutaneous Walker sarcoma (KSU), La leukemia, was transferred by intraperitoneal suspension of the клеток 1 chesky cells in physiological saline. Ino kulum contained 10® uykemic cells. The test compounds were evolved also, internally, in bruine; moreover, the first administration was carried out 3 hours after inoculation of the tumor, and then daily. Leukemia P-388 was inoculated intra-riblinically, to cells in 0.2 m ascites fluid. Compounds (Ill, 1V VII, IX) and iiniM were also administered intraperitoneally daily, starting J 3 through 1 cjTTjKH after P1ARFvivki tumor., - ;. The leukemia of the L-1210 was transferred to an anibryushiahio, to cells of its adatomy fluid into a moag, 10 cells to 6.06 W1. Physiological ecology b. Experienced - system “Internal” bus) every day ;, netsha epes t day after crossing. ; / ./, .. ,, . : Ascst cancer Zrmiha ARZ) transferred to the brain, 210 | cells in 0.06 ni of physiological saline, Fatty compounds were administered daily, starting the Circassian f Day after transplanted tumors Solid tumor - CSU, priestI, Bssec tK 6 saline solution (dissolving 1st by weight); and administered subcutaneously but ml to each animal of the Compound (tV, VI, VIt9 X) and CGPM was administered intracrically after 3. and t About days after tumor inoculation. Treatment regimens for dp of each experiment are given in Table. 6 In the trial, the doses of the newly synthesized compounds and the area of 15 mg / kg of the comparator preparation (CGNM) were equitable and amounted to 0.3 from Lfijg; relatively smaller doses were also used; doses of 20 mg / kg of compounds (tlt-V) and 40 mg / kg of compound - (X) were about 0.25 from LDg | j5, dose of 40 mg / kg of compound, (VI) was 0.2 from LD It is significant that iff in these ethics from the rays (11t-V) preparations on some of the countries were more active. The dose of the comparison drug (TSGNM) 20 kg / kg was 0.42 of LDj, and the doses of TSGNM 38 mg / kg and compounds (tV) —50 mg / kg are equivitoxic and correspond to the LD values, o of these preparations (, —the dose causing death of 10% of animals taken into experience). The criterion for antitumor cancer in leukemia and, as well as ERA, is an increase in the average life expectancy (ALE) of the experimental animals compared to the control ones, expressed as a percentage and calculated by. formula. 7. where L is the average lifetime of the living in the experimental group L 7 the average lifetime in the control group without the study day, connect. neny. in case of leukemia Lj, a “ischemic criterion for antitumor activity, which characterizes the degree of inhibition of the rate of tumor growth under the action of the test compound, was used. H and h, are the specific rates of Otl h of tumor growth in experiment and control, respectively. As an indicator of anti-tumor activity on solid tumor - CSU, inhibition (T) of an increase in tumor volume compared to control is used, expressed as a percentage and calculated by form 100 (- n where V is the average tumor volume in the control; V is the average volume tumors in the experiment. The criterion for antitumor activity is also the number or percentage of experimental, fatal, life span of which exceeded the duration of the experiment - 60 days. As can be seen from Table B, seven of the ten of the claimed compounds g (III-VI1I X ), have pronounced anti-leukemia their action: So compound (VI., V111, X) in the case of La leukemia with comparable with CGNM treatment regimens by the kinetic criterion is 3 times more effective than CGSH. Compound (VII) in its therapeutic effect on leukemia 538. P-388 is close to CGNM, and Compounds (III, IV) on this strain are more effective than TsGNM, and for (III, IV) lower relative doses were used than for TsGNM. The most effective of the proposed compounds is relative to transplanted intraperitoneally leukemia L-1210 is compound (III), Compound - (HI) at a dose of 20 mg / kg (0.26 o t LDj and identical with TsGNN treatment regimens are 1.7 times more effective than TsGNM at a dose of 15 mg / kg (0.32 from LEt). With the optimal treatment regimen by administering npeifapaTa for 2-6 days after inoculation of the tumor, compound (III) provides 100% cure for the experimental animals for the observation period - 60 days. Compounds (X) exhibit high activity against tumors grafted into the brain. In leukemia L-1210, the effectiveness of compounds (III-V) is comparable with the effect of CGNM. In the Ehrlich ascites cancer of compound (IV, V) with the same treatment regimens as in TG1III, in terms of the parameter of AFM, it is twice as effective. It should be noted that nitrosureas are one of the non-porous classes of compounds effective in the treatment of brain tumors, and among the nitrosomchevins. CGMM has the highest activity. Compound (IV) and TSGNM in equitox doses equal to (respectively 0.7 and 0.8 of LD j with similar treatment regimens) show a comparable effect on a solid tumor - CSU. Compound (VI) with a significantly lower repatriation dose-- 0.2% of LDjg inhibits tumor growth by 65% of the LCP and cures 50% of animals, and compound (VII) also at a relatively low dose — 0.3% of LD completely inhibits tumor growth and cures 70% of the experience. , UV,: EPR and mass spectra Note: For XVfl the spectrum of the liquid is given, for the rest of the substances the spectra are in Vaseline vom oil; dissolve tel - alcohol; solution in benzene, C, concentration 5..tO mol / l Chlorides of oxoamonone urea (XVIa-e) T a b and c a 2 nitroxyurea (XV a-e) T a b l and c a 3

13; ,,, / :; JESIBS.; ; ...;.; :.

-. : XnopKfiis OxoammoinYmochs (XVIft-e)

Code, i

o | shnv °

ne, V

floating

-. , : l;

ftb-3 4g «fi« ea hedgehog B3 cfe e ltca-tfiK «i 1 F« sun А xiMa Jta in .-, 7 -. - ..

XV16

90

XVI

99

91

XV1r

HU1d

92

XVI e

From MeCH-ether

Note.

Spectra in vaseline oil; The air spreads rapidly,

continuation table 3.

 V

group

eat

1617

KzgO 1563

SIR

3253, 3290

to Mr

1668

1616

1553

C-N-0

3290

N-H

1635

1617

3,280

1655

1619

1560

3280

1655 CsO

Lo

1618

1560

C-N-H

168-173

3377.3390 NH

2600 COOH

1717, 1650

& rO

1616

1560 C-N-H

"S R Yu" H

I-

 s

m and

(WITH

W, x

n t

-V

-. ::;,:. ;.,; ; ; ....,. - ..., T a in l and. C IR and UV spectra, oxoammonium and nitrourea (t-K)

a 5

59

- °

1261253 Continuation of tabl.51

“L o o o o o o o o o t 00 5; I f o (

“-Emgch .. tsK II I ill I I t about I. I i is i

 -.f,

о p- «- Ы pj w - Ч-О о n -t.

vD OxOVO iX3vC evOsOONO "CvOOvpO

tn tO.tn SOinvfi tn "L 4" in

I J. I I I I T V I T I "111

H- O -64J "-4- - 7" (((r),

o o o 1L o o o o o o o o o o o o o o o i- «t4« Mf4CM «N Me jir c JM®5TO

o o o o m o t t o o tfj No ee em o

. "IMC.0

  Cf f Pts sfs sf o cho o “F o Id yu. s $ yu 2 to ffi n m m p m y m m o o o“ “ech. (N gmm "H from 4. FJ rJ nJ P 4 bJ ft"

fe fe gj K yy fe fe y tir frT in H sgg gggg g;

about OS. - about G.

NJ: And H

and and

0)

oh oh

o - N rg N h 5;

J "1.1 II I about I I

Ltd.; $ g. to the m x. X. “s. -v,. And P P to «4 -% uv QQ 3 and I t t 7

about och

sh with

o sh cm

w

But m ..