SU1085204A1 - Preparation having cytostatic antitumoral and antireductase effect - Google Patents

Description

The invention relates to new chemical compounds belonging to the class of dipyrimidothiazines, namely derivatives of dipyrimido-5H- (4,5-b)

(4 ', 5 ^; e) (.1,4) thiazine of the general formula I

10

derivatives of dipyrimidothiazine of the formula I.

The method for producing the claimed compounds is based on the known substitution reaction of halogen in halogen derivatives of heteroaromatic compounds for residues of various amines.

Compounds of formula I are prepared by reacting a compound of formula III

where K. is β.-diethylaminoethylamino group -YN-CH ^ CH ^ CSS ^ Well) ^ ^or Ν— (Ν * - diethylcarbamoyl) .- piperazino

fifteen

N

Group

-lZk- (W (! _g 1U _g

C1

or Ν- (Ν'-methyl) piperazino group

-H ^ -CH3

20

with an amine of the general formula IV Ηί # _ζ ,

or Ν-0,4-diazabicyclo (3.3.0) nonanyl-43 group, -,

-K N-1

Derivatives of the series of dipyrimidothiazine of the general formula II are known

where K <and K ₂ is a methoxy group (11a) or K. ^ is a dimethylamino, and K ₂ is a methoxy group, or K ₍ is a methoxy group, and K-2 is a dimethylamino group.

The closest in structure and action analogue of the claimed compounds was compound On.

It was found that it has a weakly expressed cytostatic and antitumor effect. The antireductase activity of Compound Na could not be detected due to its poor solubility in water.

The purpose of the invention is the expansion of the arsenal of funds in the series of dipyrimidothiazine derivatives having cytostatic, antitumor and antireductase effects.

These properties are determined by the chemical structure of the claimed

where K ₂ is a diethylaminoethylamino group or together with a nitrogen atom Ν ~ (Ιί'25 diethylcarbamoyl) piperazine or / или'methyl) piperazine, or

Ν-0,4-diazabicyclo (3.3.0) nonanyl-43. radical in an inert organic solvent at 7C-80 ° C.

30 The starting compound of formula III is new and is a yellow crystalline substance soluble in hot lower alcohol, benzene, toluene, acetone. Mp 202-203 ⁱⁿ S.

35 A method for its preparation is based on the substitution reaction of 4,9-dioxidipyrimidothiazine with phosphorus oxychloride in the presence of dimethylaniline.

As an organic solvent, benzene or toluene is usually used.

The following examples illustrate the process for preparing the claimed compounds of formula I and the starting compound of formula III.

EXAMPLES EXAMPLE 1 Preparation of 4,9-bis (p-diethylaminoethyl) -aminodipirimido5N- (4,5-c) (4 ^g, 5'-e) (1,4) tiazyna (1a)

₅ θ To a suspension of 3 g (11.5 mmol) of 4.9 dichlorodipyrimidothiazine in 90 ml of benzene, a solution of 10.2 g (δ7 mmol) of β-diethylaminoethylamine in 30 ml of benzene is added, heated to boiling (78-80 ° С) and boiled 3 h, after which

⁵⁵ evaporated in vacuo to dryness. Triturated with water (20 ml), extracted with chloroform (3x25 ml). Chloroform is distilled off in vacuo, the residue is triturated

5

1085204

6

with petroleum ether (20 ml), the solidified substance is filtered off.

Obtain 3., 6 g (76%) of the compound '1A in the form of yellow crystals

with mp 17δ — 180 ° C (from benzene).

Found,%: C 55.46; H 7.71;

N, 30.10.

C ₂₀ H ₃₅ S _d 5 (M.M. 431.6).

Calculated,% ·. C 55.65; H, 7.70; 'ιθ N 30.30.

IR spectrum 0), cm: 3100, 3220 (4-, 5-, 9-ΝΗ).

PRI me R 2. Obtaining 4,9-bisΝ- (Ν'-diethylcarbamoyl) -piperazino-15 DIPIRIMIDO-5H- (4,5-b) (4 ', 5'-e) (1,4) thiazine (1b).

Under the conditions of Example 1, from 1 g (3.7 mmol) of 4,9-dichlorodipyrimidothiazine and 2.7 g (14.8 mmol) of Ν-diethylcar-20 bamoylpiperazine, 2 g (95%) of compound 16 were obtained as yellow crystals, with so pl. 136-142 C (from ethyl acetate).

Found,%: C 54.52; H 7, Yu; 25

N, 26.79.

С ₂₂ Н „Н ₄₁ О ₂ 5 (14 m. 569.7).

Calculated,% ·. C 54.81; H, 6.90;

N, 27.04.

IR spectrum O), cm ~ ': 1630-1640 30

(CO amide), 3200 (5-ΝΗ).

Example Z. Preparation of 4,9-bisΝ- (Ν'-methyl) -piperazinodipyrimido-5H (4,5-c) (4 ’, 5'-e) (1,4) thiazine (1c).

In the conditions of example 1 of 1 g of ₃ (3.7 mmol) of 4,9-dichlorodipyrimido-·. ^'40

thiazine and 3 g (29.6 mmol) of Ν-methylpiperazine receive 1.3 g (89.2%) of compound 1c in the form of yellow crystals with so pl. 173-174 ° C (from cyclohexane).

Found,%: C 54.06; And 6.37;

N, 31.67; 5 8.16.

C, ₈ H ₂₅ S ₉ 5 (M.M. 399.5).

Calculated,%: C 54.10; H, 6.30; hp

N, 31.5; 5 8.02.

IR spectrum 0), cm ~ ⁴ : 3200 (5-ΝΗ).

Example 4. Preparation of 4.9-bisN- ^ 1,4-diazabicyclo (3.3.0) nonanyl '4] dipyrimido-5H- (4,5-c) (4', 5 * -e) (1, 4) 50 'thiazine (1g).

4 g (29.6 mmol) are added to a suspension of 1 g (3.7 mmol) of 4,9-dichlorodipyrimidothiazine in 45 ml of benzene

1,4-diazaspirobicyclononan in 10 ml of 55 ethanol and boiled for 8 hours. The mixture is evaporated, triturated with 15 ml of water, extracted with chloroform (3x25 ml). Chloroform is distilled off in vacuo, the residue

triturated with a mixture of ethyl and petroleum ether (1: 2), left to crystallize for 72 hours. The precipitated solidified precipitate is filtered off.

Obtain 0.9 g (54.5%) of compound 1 g in the form of yellow crystals with so pl. 135-137 ° C (from ethyl acetate).

Found.%: C 58.72; H, 6.25;

N, 27.89; 5 7.4.

0 ₂₂ Η ₂₉ ν ₉ 5 (M.M. 451.6).

Calculated.%: C 58.50; H 6.40;

N, 27.90; 5 7.10.

IR spectrum O), cm: 3200 (5-ΝΗ).

PRI me R 5. Obtaining 4,9-di. chlorodipyrimido-5H- (4.5 "c) (4 *, 5’ ~ e) (1.4) thiazine (III).

a) Preparation of 4,9-dioxidipyrimidothiazine.

A mixture of 8 g (30.4 mmol) of compound Na with 400 ml of 36% HC1 is heated in a boiling water bath (95-100 C) for 2.5 hours. The precipitated substance is filtered off, washed with water (50 ml).

6.5 g (91%) of 4,9-dioxidipyrimidothiazine are obtained, which, after drying, is directed to obtain the dichloro derivative.

b) Preparation of 4,9-dichlorodipyrimidothiazine.

A mixture of 1 g (4.3 mmol) of 4,9-dioxipyrimidothiazine with 20 ml of chloro phosphorus and 1.1 g (4.3 mmol) _of dimethylaniline is boiled at 105-108 C for 3 hours, distilled off in vacuum _ about 15 ml of phosphorus oxychloride, the residue is mixed carefully while cooling with ice with 30 g of ice.

After decomposition of the residues of chloroxide, the substance is filtered off, washed with water from acid, then with alcohol.

Obtain 0.9 g (78%) of compound III in the form of yellow crystals with a so pl. 202-203 ° C (from benzene).

Found,%: C 35.53; H 1.11;

N, 26.2; C1 26.08; 5 11.15.

C ₈ H ₃ H ₅ Cl ₂ 5 (M.M. 272).

Calculated,%: C 35.30; H 1.10;

N, 25.8; C1 26.10; δ 11.80.

IR spectrum ('^), cm "^<; 3250 (5-ΝΗ).

All of the claimed compounds of formula I exhibit a cytostatic and antitumor effect. One compound 1B possesses, in addition to the above, antireductase activity.

1085204. The quantitative characteristic of the biological activity of the claimed compounds.

a) Cytostatic effect.

The cytostatic effect of the claimed compounds was studied by ϊη νίίΓο in primary cultures of normal (H) (heart of a chicken embryo) and tumor (O) (sarcoma 45) tissue in comparison with the known compound Pa.

The action of the compounds was evaluated after 72- or 96-hour in- * cubation with the test substance at '37 ° С _О The inhibition index (1 _t ) was calculated by a known method.

The results of the study are presented in table 1.

Table!

Comparative studies of the cytostatic effect of the claimed and known compounds

Connected The cult 1u (in%) at <oncent- nation pa tka- walkie-talkie mg / ml No 0.1 | 0.01 1a n 100 40 0 0 100 60 0 1b n 100 fifty 0 0 100 - 54 0 1c n 100 48 0 0 100 100 56 1g n 100 60 0 0 100 62 0 On (from - n 20 0 0 known) θ 20 0 0

As can be seen from the experimental data presented in table 1, the claimed compounds 1a-g at a concentration of 1 and 0.1 mg / ml inhibit the growth of cultures of normal and tumor tissues by 40-100%, and at a concentration of 0.1 mg / ml of the compound 1a-1g, and compound 1c and in concentration

10 ° " ⁰¹ mg / ml show selectivity of action on the tumor tissue, inhibiting its growth by 54-100 and 56%, respectively.

The known compound Na in a relatively high concentration (1 mg / ml) shows only a slight cytostatic effect on normal and tumor tissue, inhibiting its growth by 20%.

Thus, we can say that the cytostatic effect of the claimed

25 compounds are significantly higher than the well-known structural analogue of Na, and in addition, compounds 1a and 1b have a selective effect on tumor tissue.

thirty

b) Antitumor activity. The study of the activity of compounds

of formula I was carried out in experiments ϊη νϊνο in 316 rats with transplantable tumors of Jensen's sarcoma and M-1 rats in comparison with the known compound Na. For a comparative assessment of the change in tumor weight (T,%) of animals of the treated group compared with the control, the claimed compounds were administered reg. in the form of a suspension in salted., crude oil, or 'intraperitoneally (i.p.) in the form of a solution in an isotonic solution of sodium chloride once

45 per day for 8 days. The results of Ig-testing are given in table. 2.

Table 2

The results of a comparative study of the antitumor activity and toxicity of the claimed compounds

Connected Way to- Number of One-time (te Brakes Pado ^{P &} R nation denia denia rzpevtiches- growth rate one-time kaya) dose tumors Mr. Introduction mg / kg (T) nii mice mg / kg

1a

reg s

1b reg oz

8

8

20

100

47

41

150

500

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10

Compound Route of administration The number of introductions Single (therapeutic) dose, mg / kg Growth inhibition tumors (t), g at single administration to mice, mg / kg 1c in / b 8 20 58 fifty 1g reg oz 8 100 29th 500 On the famous reg oz 8 100 15-17 500

"

As can be seen from table 2, compounds 1A-g inhibit the growth of rat tumors by 29-55%. The known compound Na under the same conditions inhibits the growth of tumors by only 15-17%.

c) Assessment of the toxic effects of the compounds of formula I.

As can be seen from the data in table 2, the claimed compounds 1b and 1g are not toxic. Their lethal dose, which causes the death of 100% of the mice (bO _(O o)> with a single administration, is 500 mg / kg. ΙΛ), (οο of compounds 1a and 1c is 50–50 mg / kg.

c) Antireductase activity (inhibitory activity against dihydrofolate reductase.

Dihydrofolate reductase (DGF) was isolated from the liver of healthy animals (rats). The activity of DFR was determined by the colorimetric method according to the known method N ·

According to the technique of [Y], when studying antireductase activity, the studied compounds are used in the form of aqueous solutions with the addition of a buffer mixture.

Since the claimed compounds 1A, 1B, 1G and the analogue On are practically insoluble in water, it was not possible to detect antireductase activity in them. Of all the claimed compounds, only compound 1c was soluble in

I *

water, and therefore, only he was found antireductase activity. Compound 1c at a concentration of 1-10 ' ⁶ M inhibits the activity of DGF by 34%, and at a concentration of 1 · 1 M - by "3%.

Comparative data on the antireductase activity of the claimed compound 1c and the known Na are not given, due to the poor solubility of Pa.

As can be seen from the data of this study, the claimed compound 1B shows a pronounced antireductase activity in concentration

30 1-10 '^ M.

In conclusion, we can say that the above studies confirm the identity of the claimed properties of the compounds of formula I, namely, the presence of cytostatic and antitumor activity of all compounds in one of the compounds - 1c of an additional new property - antireductase activity.

40 So we can say

that the claimed compounds may be of interest in terms of the search for potential drugs in a number of _n-5H dipirimido "(4,5-c) F'L'-e)

45 (1,4) thiazine possessing cytostatic, antitumor and antireductase action.

Claims (3)

1.4,9-Bis-amine derivatives of dipyrimido-5H- (4,5-b) (4 ', 5' -e) (1,4) thiazine of the general formula I Κι. δ 2 where K. is β-diethylaminoethylamino, or Ν- (Ν * -diethylcarbamoyl) -piperazino-, or Ν- (ν'-methyl) -piperazino-, or N - [/ 1,4-diazabicyclo (3,3,0 ) nonanyl-42 group with cytostatic and antitumor effects. 2. 4,9-Bis-amino derivative of dipyrimido-5H- (4,5-b) (4 ', 5'-e) (1.4) thiaz.in 0 c possessing anti-reductase action, cytostatic and antitumor action. 1085204 A1 ζ 3 10 & 5204