DE2351724A1 - NEW NITROSO DERIVATIVES - Google Patents

Description

New nitrosourea derivatives

Yamanouchi Pharmaceutical Co ", Ltd.

No. 5-1, Nihonbashi-Honcho 2-chome, Chuo-ku, Tokyo (Japan)

The present invention relates to new nitrosourea derivatives of formula (II)

(ID

wherein R- and R _{2 can be the} same or different and each denotes a hydrogen atom, a lower alkyl group, a cycloalkyl group, a lower alkanoyl group or a lower alkoxy group and η has a value of 1 to 5, where n is 1, 'if R- and Rp each mean a hydrogen atom.

In the compounds of this invention having the formula (II), as stated above, η a value from 1 to 5 and Examples of the rings formed are a 2-oxopiperidine ring (n = i), a 2-oxohexahydroazepine ring (n = 2), a 2-0xooctahydroazocine ring (n = 3), a 2-oxoazacyclononane ring (n = 4) and a 2-oxoazacyclodecane ring (n-5)

R 1 and R ₂ , which may be the same or different, each represent a hydrogen atom, a lower alkyl group, a cycloalkyl group, a lower alkanoyl group or a lower one

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Alkoxy group, as already indicated above. Examples of the lower alkyl group are a methyl group, an ethyl group, an isopropyl group, a tert-butyl group and an amyl group; Examples of the cycloalkyl group are a cyclopentyl group and a cyclohexyl group. As examples of the lower alkanoyl group, there may be mentioned an acetyl group, a propyonyl group, a butyryl group and a valeryl group. Examples of the lower alkoxy group are methoxy group. an ethoxy group, a butoxy group and an amyloxy group. R ₁ and R ₂ can be arranged in any position of the ring formed

The nitrosourea derivatives of formula (II) of this invention are new connections. The nitrosourea derivatives have been used to treat leukemia and other tumors in animal tests proven to be effective and so it can be expected that these will be used as anti-tumor agents for humans and animals can.

It has been known for a long time that nitrosourea derivatives have a strong inhibitory effect against experimental ones have malignant animal neoplasms and in particular 1,3-bis (ßchlorethyl) ~ 1-nitrosourea is effective, but since such compounds are toxic, they have not yet been used or been sold.

In order to eliminate these disadvantages of the 1 ^ -Bisiß-chloräthylJ-i-nitrosourea, modifications to change its structure have been made. For example, 3-cyelohexyl-1- (β-chloroethyl) -1-nitrosourea and 3- (4-methylcyclohexyl) -1 - (β-chloroethyl) -i -nitrosourea have been synthesized, but no satisfactory results have now been obtained using these compounds obtained (see J. of Med. Chem., 2, ^Ö 92-911 (1966)).

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As the results of various researches that-the purpose had nitrosurea derivatives with strong anti-tumor effects, but to obtain low toxicity for use, the inventors have found that the compounds of this invention, Derived from. <· -Amino - ^ - lactam der ort », 60, diamino carboxylic acid have low toxicity considering their strong anti-tumor effects in animal tests.

The compounds of this invention with that already discussed Formula (II) can be obtained by nitrosotation of the urea derivatives with the formula (I)

are obtained, where R-., R ₂ and η have the same meaning as in the general formula (II).

The nitrosis is carried out in the usual way, for example it is carried out by carrying out the compound with the formula (I) "with an equimolar or excess molar amount of nitrous acid with cooling, preferably from 0 to 10 ^° C. As a solvent for the reaction is usually used an organic acid such as formic acid, acetic acid and the like, an inorganic acid such as hydrochloric acid and the like, or their solution diluted with water, The nitrous acid used in the above reaction can usually be prepared in the reaction batch. For example, 'this can be done by implementing

4 0 9 8 19/1190

one of their alkali metal salts such as sodium nitrite or one aqueous solution thereof with an organic acid such as formic acid, acetic acid and the like, or an inorganic one Acid such as hydrochloric acid, sulfuric acid and the like are prepared as a reaction solvent for the reaction will.

In addition, in such a reaction, the compound represented by the following formula

H "
NO

Jr / 1

^ X Il

IR ₂
O = C ²

NH-CH ₂ CH ₂ Cl

which has a nitroso group at the 3-position, usually educated. It is believed that such a connection arises under the influence of the amount of water in the reaction mixture. This means that if there is water in the reaction mixture is present in the reaction of the compound of formula (I) with the nitrous acid, usually a nitroso group is introduced to the nitrogen atom in the 3-position, but when the reaction of the compound of formula (I) with the Nitrous acid is carried out under anhydrous conditions, the nitroso group not in the nitrogen atom in the 3-position introduced, but is introduced to the nitrogen atom in the 1-position.

If the connection explained above with the on stick. Substance atom in the 3-position introduced nitroso group for a long time in the presence of an acid such as formic acid, acetic acid

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and the like is left to stand under anhydrous conditions, the compound is converted into the compound of the formula (II) convicted that the. Nitroso group introduced in the 1-position contains. It is preferred to carry out the reaction with cooling. Furthermore, the acid itself can be used as a reaction solvent can be used, but other suitable solvents can be used.

However, the nitrosization can also be carried out by reacting the compound of the formula (I) with nitrosyl chloride (NOCl) in acetic acid, acetic anhydride, sodium acetate or pyridine. The compound of the formula (I) _{can also be reacted with nitrogen trioxide (N 2} O ₃ ) or dinitrogen tetroxide (NgO,) in acetic acid in the presence of sodium acetate. The compound of the formula (I) can also be reacted with teutipButylnitrite (tert, -BuONO ) or tert-amyl nitrite (tert-AmONO) in hydrochloric acid or by reacting the compound of formula (I) with formyl nitrite in 100% formic acid.

The compounds of these represented by the formula (II) Invention can be isolated and purified by methods known per se, for example by extraction, recrystallization, Column chromatography and the like.

The starting material for this invention having the formula (I) includes the stereoisomers, the dl form and the optically active d-form and 1-form and if these are used as the starting material the established connection will be with the ent-. speaking steric or optical configuration. Likewise, if a mixture of the isomers is used as starting material in the reaction, the reaction mixture can through fractional crystallization, column chromatography and the like can be separated.

409 8 19 /! 190 _{0R1G (NÄL 1NSPBCTE} d

In addition, it should be noted that the starting material of the formula (I) represents a new compound and, for example, by converting the lactam compound to the formula

(CH ₀ )

2'n

into the corresponding 3-halo compound according to the method of Juli et al. (J. Org "Chem., $ 2, 2425 (1964) and the method according to Robert J, Wineman et al. (J. Am. Chem. SOC ₁ OEO 6233 (1953)) by converting the product into a 3- Azalactam compound according to the method of M. Brenner et al. (HeIv. Chira. Acta, / ^ L, 1Ö1 (195Ö)), catalytic reduction of the 3-azalactam compound according to a known method to give the 3-aminolactam compound with the formula

to form, in order to then umzu the product with ß-chloroethyl isocyanate put, can be obtained.

The antitumor activity LD ₁ Q ^^ "^ 50 ^ ⁶¹ * compounds of this invention shall now be demonstrated by the following Studies I and II.

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Examination I (Crocker's Sarcoma 100)

Small pieces of tumor tissue, "1.5 nmr in size, were implanted subcutaneously in the right axillary region of ddn mice (", 10-22g). Ten mice each form a group. 24 hours after implantation, the compounds (dissolved in aqueous solution containing 2% (+1), 3% (+2) or - \ & (+3) in ethanol, except for the compounds marked with (> ^) ') injected intraperitoneally only once. At the end of the second week after implantation, the number of deaths and tumor diameter were calculated using the formula axb, where a and b are the larger and smaller diameters, respectively. the results are shown in Table I.

The tumor index {%} was calculated using the formula T / C χ. C means the mean control tumor size. T means the mean treatment tumor size.

The results of the "treatment in the table have been taken through the following symbols and meanings as specified by K. Sugiura (Cancer-Research, .22, (2), 179 (1967)).

Symbol,

Degree of tumor Tumor index Inhibition • no 76-100 easy 51-75 middle 26 - 50 clear β - 25 Completely 0-7

40981.9 / 1 1 90

1-ifl-chloroethyl) - 13.0 28.6 2 351724 3- (1-methyl-2-oxo- 16.9 37.2 Table 1 3-hexahydroaz epinyl) -22.0 48.4 Result at the end of the second week Compound dose 1-nitrosourea 7.7 Tumor result of the number of (mg / kg) 10.0 index treatment Kill 13.0 56.2 + 0/10 1- (ß-chloroethyl) - 16.9 50.0 + 0/10 3- (5-methyl-2-oxo-3- 22.0 10.9 '++ 0/10 hexahydroazepinyl) - 26.6 3.4 ⁺⁺⁺ 0/10 1-nitrous raw material 7.7 < _t 3 +++ 0/10 +2 10.0 0/10 13.0 59.7 + 1/10 i- (ß-chloroethyl) - 16.9 46.2 + 0/10 3- (5-methyl-2-oxo-3- 22.0 25.9 ++ 0/10 hexahydrο az epinyl) - 28.6 O 1 ι ι ι
JJ, 4 ^ ⁺ + 1/10 1-nitrous urea f 7.7 0/10 +3 10.0 1.0 +++ 0/10 13.0 73.7 ± 0/10 1- (B-chloroethyl ) - 16.9 70.5 + 0/10 3- (2-oxo-3-piperi- 22.0 8.2 4+ 0/10 dinyl) -1-nitroso- 28.6 7.4 +++ 0/10 urea 1.3 +++ 0/10 +1 1 8 ι j ι 1/10 7 O ι ι ι 0/10 0.2 +++ 0/10 \ J m fc ITT 0/10 0.6 +++ 0/10 0.3 +++ 0/10 0.2 +++ 0/10

The compounds (dissolved in aqueous solution containing efo ethanol and % polyethylene (60) -hydrogenated castor oil derivative) were injected.
(According to this, the character () *!) And the preceding markings

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(+1, +2, +3) have the same meaning.

Investigation II

The compounds were administered by a single subcutaneous injection to ddN mice (6, 12-22 g). Each group consisted of ten mice. By determining the number of deaths 60 days after the administration of the compounds, the LDjQ and LD ^ ₀ values were obtained. The results obtained are shown in Table II. ·

Table II. .

Compound LD _1Q (mg / kg) LD ₅₀ (mg / kg)

1- (ß-oleoethyl) -3- (7-methyl-

2-oxp-3-hexahydroazepinyl) -1- 39 _s 5 45.0

nitrosourea Λ "■

1- (ß-chloroethyl) -3-ί 5 ~ methyl-

2 «oxo-3-hexahydroazepinyl] -1-23.3. 32.6 nitrosourea +2

1- (ß-chloroethyl) -3- (5-methyl-

2-oxo-3-hexahydroazepinyl} -1- 24.9 31.7

nitrosourea +3

1- {ß-chloroethyl) -3- (2-0x0-3-

piperidinyD-i-nitroso urine »21.4 27.2

fabric +1

example 1

a) in 30 ml of chloroform was 1.15 g of 3-amino-2-oxopiperidine dissolved. After 10 ml of chloroform solution, which contained 1 ^ 2 g of ß-chloroethyl isocyanate, were added dropwise to the Solution with stirring at 0-5 ° C, the mixture became overnight stirred at room temperature. The crystals thus formed were separated by filtration and removed from methaol

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ORIGINAL INSPECTED

recrystallized. 1.52 g of 1 ~ (β-chloroethyl) -3- (2 ~ oxo-3-piperidinyl) urea were obtained with a melting point of 106 107.5 ° C received.

Elemental analysis for ( Ί Tl , N ₃ O ₂ Cl: H
6th , 44 NC
19th , 13 Calculated: c (i
43 %)
, 73 5 , 93 19th , 22 Found: 43 , 76

_{b) T 1} J g of 1- (ß-chloroethyl) -3- (2-oxo «3-piperidinyl) urea was dissolved in 40 ml of 99 # formic acid and then 0.7 g of sodium nitrite was added in small portions over a period of 30 Added to the solution with stirring at 0-5 ° C. The mixture was stirred at 10-13 ° C. for a further 3 days. To the mixture was added 00 ml of ice water and the product was extracted three times with 40 ml of chloroform each time. The extracts were combined, washed three times with water, dried over anhydrous sodium sulfate, and then chloroform was distilled off under reduced pressure. The residue obtained was dissolved in a small amount of chloroform. A petroleum ether-acetone solvent mixture (5: 1 by volume) was added to the chloroform solution. The crystals formed were separated by filtration. 1- (β-chloroethyl) -3- (2-oxo-3 ~ piperidinyl) -1-nitrosourea was obtained. The filtrate was concentrated and a small amount of acetone and petroleum ether were added to the concentrate. The batch was left to stand to crystallize out. The crystals were filtered off and further 1- (ß-chloroethyl) -3- (2-oxo-3-piperidinyl) -1-nitrosourea with a melting point of 123-126 ° C was obtained.

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Example 2.

6 g of 1- (β-chloroethyl) -3- (2-oxo-3-piperidinyl) urea were added to 160 ml of 99% formic acid dissolved, and after more gradually Adding 2.46 g of sodium nitrite to the solution over the course of 30 minutes with stirring at 7 ° C., the batch was continued for 5 days stirred at 5 - 7 ° C. When the implementation was finished, that was The reaction product was dried by freeze-drying and the residue obtained was washed three times with 70 ml of chloroform extracted. The extracts were combined, three times with sat aqueous sodium chloride solution and washed once with cold water. It was made over anhydrous sodium sulfate dried, and then the solvent was distilled off under reduced pressure. To the residue obtained acetone and a small amount of petroleum ether were added. The crystals formed were separated by filtration. 3 g of 1- (β-chloroethyl) -3- (2-oxo-3-piperidinyl) -1-nitrosourea were obtained with a melting point of 123-127 ° G obtain.

Elemental analysis for CgH.-Ν, O ₃ Cl:

C (fo) H(#) N (#) Calculated: 30.64 5.27 22.53 Found: 3ö, 5ö 5.31 22.50 Example 3

a) In 20 ml of 99% formic acid, 1.1 g of i- (ß-chloroethyl) -3- (2-oxo-3-piperidinyl) urea was added dissolved, and then ml of an aqueous solution of 0.7 g of sodium nitrite dropwise added to the solution with stirring at 0-5 ° C. After further stirring the mixture for 30 minutes was 50 ml of water are added, creating an oily material

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ORIGINAL INSPECTED

was formed. The oily material was extracted twice with 10 ml of chloroform each time. The extracts were combined, Washed twice with 5 ml of water each time and over anhydrous Sodium sulfate dried * The solvent was reduced under reduced Distilled pressure. The obtained oily matter was dissolved in a small amount of methanol and the solution was left to cool, whereby crystals were deposited. The crystals were collected by filtration severed. There were 0.05 g of 1- (ß-chloroethyl) -3- (2-oxo-3-piperidinyl) -3 * -nitrosourea obtained with a melting point of 154-156 ° C.

Elemental analysis for CgHj ~! LO ~ Cl:

. c (f.) Calculated: 30.64 Found: 30.51

N (Ji)

5.27 22.53 4; dÖ 22.75

b) 3.5 g of 1-β-chloro-ethyl) -3- (2-oxo-3-piperidinyl) -3-nitrosourea were dissolved in BO ml of 99-100% formic acid. The solution was left to stand for three days at 10-13 ° C., then 300 ml of ice water was added to the solution and the product was extracted three times with 70 ml of chloroform each time. The chloroform extracts were combined, washed three times with water, dried over anhydrous sodium sulfate, and then chloroform was distilled off under reduced pressure. The residue was dissolved in a small amount of chloroform, a petroleum ether-acetone mixed solvent (5: 1 by volume) was added to the solution, and the crystals formed were separated by filtration. The crystals were dissolved in chloroform, a small amount of petroleum ether was added to the solution, and the crystals formed were filtered off. The filtrate was concentrated and after adding acetone and a small amount

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ORfGINAL INSPECTED

Petroleum ether to the concentrate was the approach calmly. The crystals formed were separated off by filtration and 1- (β-chloroethyl} -3- (2-oxo-3-piperidinyl) -1-nitrosourea was obtained with a melting point of 123 - 126 ° C get%

Element ar analysis for CgH- -N.OoCl:

'; C (%). ■ ü {fo) H (fo)

Calculated: 30.64 5.27 22.53 Found: 30.49 5.18 22.49

Nuclear Magnetic Resonance Spectrum (CDCIoKi

^r d f 7.95 (IH, d, J = 7.1 Hz, H of the nitrogen atom in the 3-position of the urea, '

4 * 45 (1H, m, H in the 3-position of the piperidinyl group,

4jii4 (2H, t, J = 6 Hz, H of the nitrogen atom in the 1-position of the urea bonded methylene group.

3j49 (2H, t, J = 6 Hz, H of the bonded to the chlorine atom Methylene group.

Then 20.5 mg of the product in 10 ml of dry dichloromethane dissolved and added 43 mg of cyclohexylamine. The mixture was refluxed with heating for 3 hours. Then the solvent was distilled off under reduced pressure and ether was added to the residue. The crystals formed were separated by filtration. 25 mg of 1-cyclohexyl - ^ - (2-oxo-3-p: lperidinyl) were urinated off with a melting point of 245-249 ° C.

Elemental analysis for C ₁₉ H ₉₁ NO ₅ :

Calculated: A. 0 60 , 22 1 1 9 8th 0 , 85 17th , 56 Found: 60 , 19th 8th , 79 17th , 50 98 19 /

ORIGINAL INSPECTED

Mass analysis: M = 239

In addition, 1- (ß-chloroethyl) -3- (2-oxo-3-piperidinyl) -3-nitrosourea was obtained by reacting the starting material with cyclohexylamine, 1 ■? (ß-chloroethyl) -3-cyclohexylurea obtained with a melting point of 130 ° C. (J, Med. Chem., £, Ö92 (1966)).

Example 4

a) 3,4 3-Amino-6-methyl-2-oxopiper.idin (a mixture of 1 ί 1 cis-form and trans-form) was dissolved in 40 ml of dry chloroform. Then 10 ml of a dry chloroform solution was added dropwise which 3.1 g of .beta.-chloroethylisocyanate containing added under cooling with ice water and the ^g emic stirred 2 hours. After completion of the reaction, the solvent was distilled off from the reaction product under reduced pressure. Then, ether was added to the residue, whereby crystals were formed. The crystals were separated by filtration and recrystallized from acetone-benzene. It was 5.0 β 1- (ß-chloroethyl) -3- (6-methyl-2-oxo-3-piperidinyl) urea (a mixture of 'lit cis form and trans form) with a melting point of 149 - 151 ° C obtained.

Elemental analysis for C ₉ H- ^ N ₃ O ₂ Cl

c (f ₃ ) H (Ji) 15.41 CKf ₀ ) Calculated: 52.04 7.02 15.39 12.99 found: 52.70 7.00 12.03

b) 1.2 g of 1- (β-chloroethyl) -3- (6-methyl-2-oxo-3-piperidinyl) urea were added to 40 ml of 99% formic acid added and then 0.7 g of sodium nitrite was slowly added to the solution

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Added over 30 minutes with stirring at 0-5 ° C. After further stirring the mixture for 5 days at 10-13 ° C, 10-0. ml of ice water was added to the mixture. The product was then extracted 3 times with 50 ml of chloroform each time. The extracts were combined, three times with Washed water, dried over anhydrous sodium sulfate and the chloroform was distilled off under reduced pressure.

Acetone and a small amount would be added to the residue obtained • Add the amount of petroleum ether and the crystals that have formed separated by filtration and recirculated from chloroform. It was 1- (ß-chloroethyl} -3- (6-methyl-2-oxo ~ 3 ^ piperidinyl) -1-nitrousourea obtained with a melting point of 157-159 ° C.

Example 5

a) In 25 ml of dehydrated chloroform was 3, Ö S 3-amino-5-methyl-2-oxohexahydroazepine dissolved. Then a solution of 3.1 g of ß-chloroethyl isocyanate in 10 ml was dehydrated Chloroform added dropwise to the solution with stirring 5 - 10 ° C added. After stirring the mixture for two hours at room temperature, the crystals formed became separated by filtration. 2.1 g of 1- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) urea were obtained obtain. After washing the filtrate with water, the filtrate was dried over anhydrous sodium sulfate, and that Solvent was reduced from the residue under reduced pressure Distilled pressure. Treatment of the oily residue obtained with acetone and petroleum ether gave a further 1.4 g 1 - (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) urea obtain. The overall yield of the product was 3.5 g with a melting point of 1Ö3 - 104 ° C (with decomposition

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INSPECTED

Settlement.

Elementary analysis for

'C (56) H (SO Where) Calculated: 40.49 7.32 16.96 Found: 40.42 7.27 16.59

b) · In 40 ml of 99% formic acid, 2.9 g of i- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroaz_epinyl) urea and then 3.4 g of sodium nitrite were slowly added with stirring 5 - 7 ° ^{C added} to the solution. After stirring the mixture further for 2 hours at the same temperature as indicated above, 250 ml of water was added to the mixture, whereby a viscous oily product was formed. The oily product was extracted three times with 50 ml of chloroform each time. The extracts are combined, dried over anhydrous magnesium sulfate, and then chloroform was distilled off under reduced pressure. The viscous oily residue thus obtained was dissolved in ethyl acetate and then petroleum ether was added to the solution. The. Crystals formed in this way were separated by filtration. 1.1 β .1- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea with a melting point of 110-119 ° C. was obtained (with decomposition) obtained.

Elemental analysis for C ₁₀ H ₁₇ N ₄ 0 ₃ Cl: H( 19th NC %): c ($ ß) 6, 30th 20th , 25 Calculated: 43.40 6, 20th , 20 Found: 43.67

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- 17 - Example 6

a) In 50 ml of chloroform 1.0 g of 3-amino-7-methyl-2-oxo-. hexahydroazepine dissolved. Then a solution of 1.5 g

ß-chloroethyl isocyanate in 20 ml of chloroform is added dropwise added to the solution with stirring at 0-5 ° C. To further stirring the mixture for one hour at room temperature, the product was washed twice with 20 ml of water each time, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The crystals thus formed were separated and recrystallized from acetone. It became 1.8 g 1 - (β-chloroethyl) -3- (7-methyl-2-oxo-3-hexahydroazepinyl} urea obtained with a melting point of 200-209 ° C.

Elemental analysis for C. Jf

Calculated: .40.49 7.32 16.96. Found: 40.20 7.21 17.10

b) 1.7 g of i- (β-chloroethyl) -3- (7-methyl-2-oxo-3-hexahydroazepinyl) urea were added to 20 ml of 99% formic acid dissolved and then 2 g of sodium nitrite was slowly added to the solution in small portions' within 30 minutes with stirring 0 - 5 ° C added. After further stirring the mixture for 2 hours at the same rate as indicated above Temperature - 200 ml of water was added, whereby light yellow crystals were formed. The crystals were separated by filtration and washed with water. There was 1.4 δ 1- (ß-chloroethyl) -3- (7-methyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea obtained with a melting point of 133-134 ° C.

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Elementary analysis for C ^ HjgN.OoCIi

Calculated: 43.40 7th 6th , 19th 20th , 25 Found: 43.51 6th , 25 20th , 12 example

a) 0.76 g of 3-amino-O-methyl-2-oxooctahydroazosin was added to O ml of chloroform dissolved. Then a solution of 0.6 g of β-chloroethyl isocyanate in 2 ml of chloroform was added dropwise added under cooling. The mixture was further stirred for 1.5 hours at room temperature. After the implementation ended, the reaction product was concentrated under reduced pressure. Ether became the concentrate obtained was added, and the crystals formed were separated by filtration. 0.92 g of 1- (ß-chloroethyl) -3- (ß-methyl-2-oxooetahydroazosin-3-ylurea were obtained with a melting point of 207 - 210 ° C ..

Elemental analysis for C ₁ -

Calculated: 50.40 7.70 16.05

Found: 50.39 7.03 16.12

b) To Ö ml of 99% formic acid, cooled to 10 ° C, was added 0.250 g 1- (ß-chloroethyl) -3- (Ö-methyl-2-oxooctahydroazosin-3-yl) urea added. Then, drop by drop, became a Solution of 0.13 g of sodium nitrite in 1.2 ml of water was added to the mixture while cooling to 10 ° C. The mixture was Stirred for 10 minutes at the same temperature and then cooled to 0 ° C. Then 56 ml of ice water was added to the mixture was added and the product was washed with chloroform

409819/1190

extracted. The extract was washed over with water dried anhydrous sodium sulfate and concentrated under reduced pressure at room temperature. To the received Acetone and ether were added to the residue, and the mixture was allowed to stand. The resulting crystals were separated by filtration. There was 0.17 g of 1 - (ß-chloroethyl) -3- (Ö-methyl-2-oxooctahydroazosin-3-yl) -1-nitrosourea with a melting point of 129 - 130 ° C

obtain. C ₁₁ H ₁₉ N ₄ C ö L: H (St) 19th , 27 Elemental analysis for C (St) 6.59, 19th , 30 45.44 6.61 Calculated: 45.49 Found: example

a) 2.7 g of 3-amino-7-butyl-2-oxohexahydroazepine were dissolved in 70 ml of anhydrous chloroform. A solution of 2.4 g of β-chloroethyl isocyanate in 30 ml of chloroform was then added dropwise to the solution at 0 ° -5 ° C. with stirring. The mixture was stirred for 2 hours. The reaction product was then washed twice with 30 ml of water each time and dried over anhydrous sodium sulfate. The solvent was then distilled off under reduced pressure, and the obtained liquid product was subjected to silica gel column chromatography. By developing the product using a 19 x 1 volume ratio of a solvent mixture of ethyl acetate and methanol as the eluate, 2 g of crystalline 1- (β-chloroethylM - ^ - butyl ^ -oxo ^ -hexahydroazepinyl urine and 0.9 β paste were obtained.
The melting point of the crystals was 153-154 ° C and the

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The Rf value of the product purified in this way was after Silica gel thin layer chromatography using a solution of ethyl alcohol mixed in a volume ratio of 9: 1 and methanol 0.5.

Elemental analysis for C. "H ₂ , N ~ O ₂ C1:

C (%) H (ji) N (Ji)

Calculated: 53, BB 3.35 14.50 Found: 53.76 δ, 44 14.71

On the other hand, the Rf value of the paste was found by silica gel thin layer chromatography using a 1: 9 volume ratio of a mixed solvent Chloroform and ethyl acetate 0.45

Elemental analysis for C

C {fo) n (%) N (#). Calculated: 53, BB 8.35 14.50 Found: 53.69 0.21 14.31

b-1) In 7 ml of 99% formic acid, 0.46 g became more crystalline 1- (β-chloroethyl) -3- (7-butyl ~ 2 ~ oxo-3-hexahydroazepinyl) urea dissolved. Then 0.43 g of sodium nitrite-f slowly added to the solution with stirring at 5-10 ° C. After further stirring the mixture for 2 hours at the same temperature already mentioned above 30 ml of cold water was added, causing white turbidity to appear and a semi-solid material to be formed. The product was extracted three times with 20 ml of chloroform each time. The extracts were combined, twice with 20 ml of water each time washed and then dried over anhydrous sodium sulfate. Then chloroform was distilled off under reduced pressure and the solid product obtained became off

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Recrystallized acetone petroleum ether to give 0.4 g of crystalline 1 - (β-chloroethyl) -3- (7-butyl-2-oxo-3-hexahydroazepinyl) -1 nitrosourea with a melting point of 117 - 119 ° C to obtain. ""

The Rf value of the product on silica gel thin layer chromatography using a 1: 9 volume ratio of a solvent mixture from'Chloroform and ethyl acetate was 0.75.

Elemental analysis for C ₁ ^ H ₂ ^ N.0 ^ Cl:

H (#) M (SC) Calculated: 4 [beta], 9 & 7.27 17.57 Found: 40.79 7.07 17.35

b-2) In 7 ml of 99% formic acid, 0.5 g became more pasty 1 - (β-chloroethyl) -3- (7-butyl-2-oxo-3-hexahydroazepinyl) urea dissolved. Then 0.43 g of sodium nitrite was slowly added to the solution with stirring at 5-10 ° C. The product was then, as indicated in step b-1, further treated. 0.4 g of oily 1- (β-chloroethyl) ~ 3- (7-butyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea was obtained obtain.

The Rf value of the product was found by silica gel thin layer chromatography using a 1: 9 volume ratio of a mixed solvent of chloroform and ethyl acetate 0.62.

Elemental analysis for C ₁ -Hp-JN, 0.Χ1

Calculated: 4Ö, 9Ö 7.27 17.57 Found: 40.73 7.52 17.61

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Example 9

a) In 20 ml of anhydrous chloroform was 3.5 S $ -amino-5-methyl-2-oxohexahydroazepine (The picrate of the compound showed the melting point at 221 ° C) dissolved. To the solution a solution of 2.9 g of ß-chloroethyl isocyanate was added dropwise in 10 ml of anhydrous chloroform with stirring at 5 - 10 ° C added. The mixture was left for 4 hours at room temperature stirred, whereby crystals were formed. The crystals were separated by filtration. It was 3.7 g of 1- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroaze-. pinyl) urea obtained. The filtrate was washed with water and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, whereby crystals were formed. The crystals were separated by filtration. The crystals turned out Recrystallized acetone. 0.0 g of i- (β-chloroethyl) -3- (5-raethyl-2-oxo-3-hexahydroazepinyl) urea was obtained with a melting point of 1Ö2 - 103 ° C (with decomposition). The total amount of the product obtained was 4.5 S

Elemental analysis for C-q

C {%) H(%) N (JS) Calculated: 40.49 7.32 16.96 Found: 40.45 7.30 16.7-5

b) In 75 ml of 99% formic acid, 4.3 g of i- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) urea was added (produced dissolved according to step a) above »Then 3.46 g of sodium nitrite was slowly added to the solution with stirring at 5-6 ° C added. After further stirring the mixture for 4 hours at 3-5 ° C., 150 ml of water was added to the mixture was added, thereby forming an oily material.

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The oily material was washed three times with 50 ml of chloroform each time extracted, the extracts were combined, dried over anhydrous Magnesium sulfate, and then chloroform was distilled off under reduced pressure. The so-obtained oily product was dissolved in a small amount of ethyl acetate. Then petroleum ether was added to the solution and the mixture was frozen in a cryogenic freezer

, cooled to form light yellow crystals. By separating the crystals by filtration, 2.6 g of 1- (β-chloroethyl) -3- (5-methyl-2-Qxo-3-hexahydroazepinyl) -1-nitrosourea were obtained with a melting point of 11 β - 119 ° C (with decomposition) obtained. .

Elemental analysis for Cj ₀ H ₁ J * 1.Q-Cl;

Calculated: 43.40 6.19 20.25 Found: 43.5S 6.29 20.21 Example 10

a) Araino-5-methyl-2-oxohexahydroazepine was dissolved in 15 ml of anhydrous chloroform (the compound had a melting point of 165 - 166 ° C). A solution of 1.0 g ß-chloroethyl isocyanate in 10 ml of anhydrous chloroform was added dropwise to the solution with stirring at 5-10 ° C. The mixture was 4 ^s HOURS at RäumtempeJratuir stirred, whereby crystals were formed. The crystals were separated by filtration. 3.2 g of 1- (β-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) urea with a melting point of 147-1.49 ° C. were obtained.

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Elemental analysis for

Calculated: 40.49 7.32 16.96 (Findings: 4 ^, 51 7> 35 16.70

b) 3.0 ζ 1- (ß-chloroethyl) -3 - (5-methyl-2-oxo-3-hexahydroazepinyl) urea (prepared in step a above) was dissolved in 50 ml of 99% formic acid. Then slowly 2.42 g of sodium nitrite was added to the solution with stirring at 5 - 6 ⁰ C added. The product was treated further as indicated in example 9b. 1.4 £ 1- (B-chloroethyl) -3- (5-methyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea with a melting point of 111-112 ° C. (with decomposition) was obtained.

Elemental analysis for C-qH- "N, OqCI:

Calculated: 43.40 6, 19th found: 43.49 6, 25th Example 11

21;

a) In 45 ml of anhydrous chloroform, 4 g of 3-amino-5 * " cyclohexyl-2-oxohexahydroazepine dissolved. Then became the Solution with stirring and cooling a solution of 2.5 g of ß-chloroethyl isocyanate in 10 ml of anhydrous chloroform added dropwise to the solution. After that, that became

Mixture stirred for 3 hours at room temperature. The crystals formed were separated by filtration. It 1.5 g of 1- (β-chloroethyl) -3- (5-cyclohexyl-2-oxo-3-hexahydroazepinyl) urea was added obtain. The filtrate was washed with water, dried over anhydrous magnesium sulfate , and then the solvent was reduced under

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Pressure distilled off * By recrystallization of the residue from acetone, 1.7 g of 1- (β-chloroethyl) -3 ^ (^ cyclohexyl-2-oxo-3-hexahydroazepinyl} urea became obtained · The total yield of the product was 3.2 g. Melting point 195 ° C (below Decomposition).

Elemental analysis for C-cHg ^ NoOgGLi

C (SS) -H (Si) N (Si) Calculated: 57.04 8.30 , 13.30 Found ι 57.12 8.27 13.32

b) 1.55 β 1- (β-chloroethyl) -3- (5'-cyclohexyl-2-oxo-3-hexahydr'oazepinyl) urea was dissolved in 30 ml of 99% formic acid. Then 1.02 g of sodium nitrite was slowly added to the solution with stirring at 5-6 ° C. After stirring the mixture further for 4 hours at 0-5 ° C, 60 ml of water was added, thereby forming a viscous, resinous material. The product was extracted three times with 30 ml of chloroform each time. The extracts were combined, washed with water and dried over anhydrous magnesium sulfate. By distilling off the solvent under reduced pressure and treating the syrupy residue with a small amount of ether, 1.12 g .1- (β-chloroethyl) -3- (5-cyclohexyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea was obtained obtained with the melting point 134 ° C (with decomposition).

Elemental analysis e for ^G 15 U T *, O-Cl: H( ^) N (Ji) c (: 7, 31 16.25 Calculated: 52 & 7, 33 16.23 Found: 52 , 21

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Example 12

a) In 70 ml of anhydrous chloroform was-2,7 β 3-amino-5 * - Dissolved methoxy-2-oxohexahydroazepine .. Then a solution of 2.4 g of ß-chloroethyl isocyanate was added dropwise to the solution added in 30 ml of anhydrous chloroform. The mixture was stirred for 30 minutes, giving the solution became cloudy and crystals were formed. After stirring the batch for a further 2 hours, the Separated crystals by filtration and washed with 10 ml of chloroform and 10 ml of ether. It was 3.3 g of 1- (ß-chloroethyl) -3- ( 5 ~ methoxy-2-oxo-3-hexahydroazepinyl) urea with a melting point of 10 ° -109 ° C.

Elemental analysis for C. qH

Calculated: 45.54 6, ÖÖ 15.93 Found: 45.72 6.71 15.79

b) In 50 ml of 99% formic acid, 3.3 g of i- (β-chloroethyl) -3- {5-1 ^ 110X3 ^ 2-0X0-3-116X3113 ^ 0azepinyl) urea dissolved. While stirring the solution at 4- # ° C, 3.5 3 sodium nitrite slowly added to the solution over the course of an hour Further stirring of the mixture for one hour, 260 ml of cold water were added to the reaction product, and the product was extracted three times with 70 ml of chloroform each time. The extracts were combined, three times with each Washed 100 ml of water, then saturated with an aqueous Washed sodium chloride solution and then dried over anhydrous sodium sulfate. By distilling off Of the chloroform from the product under reduced pressure, 3.5 g of a viscous residue was obtained. The viscous one The residue that was obtained was subjected to silica gel column chromatography, and it was washed with a solution

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Medium mixture of ethyl acetate and chloroform in volume ratio Developed 1: 1 as eluate. 2 g of crystalline 1- (β-chloroethyl) -3- (5-methoxy-2-oxo-3-hexahydroazepinyl) -1-nitrosourea were obtained with a melting point of 131 - 132 ° C ..

Elemental analysis e for C ₁₀ H ₁₇ N 4 ° 4 ^C1 13th •
• H (JC) "" N (Ji ) 5th Ö5 19 14th Calculated: 41. - 5.59 19 20th Found: 40, I) »03 .. , 92 example

a) In 30 ml of 99% formic acid, 1.74 g of 1- (β-chloroethyl) -3- (6-methyl-2-oxo-3-piperidinyl) urea was added (1: 1 mixture of the cis form and the trans form) dissolved »After the solution has cooled down a solution of 1.05 g of sodium nitrite in 12 ml of water was added dropwise over 5 minutes to 10 ° C added. The mixture was stirred at 0-7 ° C. for a further 25 minutes. 125 ml of ice water were added to the batch and then it was extracted three times with 30 ml of chloroform each time. Then, after drying the chloroform extract over anhydrous sodium sulfate, chloroform was added distilled off under reduced pressure. The residue was dissolved in acetone. Then the solution became a small one Amount of petroleum ether added. The resulting solution was left to stand. The crystals then deposited were separated by filtration. 1.40 g! - (ß-chloroethyl ) -3- (6-methyl-2-oxo-3-piperidinyl) -3-nitrousourea obtained with a melting point of 156 ° C.

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Elemental analysis for CqH ^ N.O ^ GI:

G (fo) H(#) N (ji). Calculated: 41.15 5.75 21.34 Found: 41.20 · 5, Ö3 21.30

b) In 40 ml of 99% formic acid, 1.40 g of 1- (β-chloroethyl) -3- (6-methyl-2-oxo-3-piperidinyl) -3-nitrosourea was added dissolved. Then the solution was left to stand for 5 days at 10-13 ° C. permit. 200 ml of ice water was added to the solution. and then the reaction product was three times with 70 ml Chloroform extracted. Then, after drying the chloroform extract over anhydrous sodium sulfate, chloroform distilled off under reduced pressure. Thereafter, acetone and a small amount of petroleum ether were added to the residue added. When the mixture was left to stand, crystals were precipitated, and these were separated by filtration. · Recrystallization of the crystals obtained from chloroform gave I- (β-chloroethyl) -3- (6-methyl-2-oxo-3-piperidinyl) 1-nitrosourea with a melting point of 157-159 ° C obtain.

Element ar analysis for CgH ₁ , -Ni 0.X1:

G (fo) H (%) N (*).

Calculated: 41.15 5.75 21, '34

Found: 41.09 5.01 21.22

Nuclear Magnetic Resonance Spectrum (Dg-DMMSO):

1s>: S, 3Ö (1H, d, j = ÖH _z , H of nitrogen in the 3 ~ position of urea),

4.3 (1H, m, H in the 3-position of the piperidinyl group) 4.10 (2H, t, J = 6H _z , H in the methylene group connected to the nitrogen atom in the 1-position of urea) 3.67 ( 2H, t, J = 6h, H in the methylene group attached to the chlorine atom).

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Example 14

a) 93 mg of N-acetyl-3-amino-2-oxohexahydroazepine were added to 2 ml of anhydrous chloroform and given 41 mg of triethylamine while cooling with ice. Then 1 ml of chloroform, which Containing 47 mg of B-chloroethyl isocyanate, to the mixture below Stir and ice cooling added. Then, after stirring for an additional hour, the mixture was cooled to room temperature. After the reaction had ended, the solvent was distilled off from the reaction mixture under reduced pressure. The residue obtained was recrystallized from a mixture of acetone and ether. 90 mg of 1- (ß-chloroethyl) -3- (N-ac etyl-2-oxo-3-hexahydroaz epinyl) urea with a melting point of 177-179 ° C.

Elemental analysis for C ^^ H

47.92 H(*) H (Jl) Cl (^) Calculated: 47, ÖÖ 6.5 * 15.24 - 12.06 Found: 6.39 15.09 12.77

b) In 3.06 ml of 99% formic acid, ÖÖ mg of 1- (ß-chloroethyl) -3- {N-acetyl-2-oxo-3-hexahydroazepinyl) urea was added dissolved and then 23 mg of sodium nitrite was slowly added, with stirring, to the solution at 7-10 ° C. Then became after Stir the mixture for 20 minutes at the same temperature, adding 100 ml of ice water to the reaction mixture. The product was mixed three times with 10 ml, "20 ml and then extracted 20 ml of chloroform. The extracts were combined and then the combined chloroform extracts were washed with water, "dried, .and chloroform was under distilled off under reduced pressure. The oily obtained Material was obtained from a silica gel column chromatography-lower threw. It was developed with chloroform. It became 60 mg

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oily 1- {ß-chloroethyl) -3- (N-acethyl-2-oxo-3-hexahydroazepinyl) -1-nitrosourea obtain.

The Rf value of the product on silica gel column chromatography using a mixed solvent of chloroform and η-hexane in a volume ratio of 5: 1

Elemental analysis for C. j H. "N, 0, Cl:

cm H(%) ν m Cl (Jt) Calculated: 43.36 5.63 10.39 11.63 Found: 43.39 5.64 10.43 11.60

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Claims (6)

~ 31 - Claims
1, nitrosourea derivatives with the formula NH-I OMC. "2
I. N-CH ₂ CH ₂ Cl NO wherein R- and R _{2 are the} same or different and each represents a hydrogen atom, a lower alkyl group, a cycloalkyl group, a lower alkanoyl group or a lower alkoxy group and η is 1-5, where η is 1 has, R- 'and R ₂ each represent a hydrogen atom. 2. 1- (β-chloroethyl) ^ 3- (2-oxo-3-piperidinyl) -1-nitrosourea according to claim 1. 3. Process for the preparation of the nitrosourea derivatives mentioned in claim 1, characterized in that a urea derivative with the formula I Jy O = C ^(CH 2> n NHCH ₂ CH ₂ Cj 409819/1190 where R., Rp and η have the meaning already mentioned, nitro si er t * <becomes. 4. The method according to claim 1 for the preparation of 1- {ß-chloroethyl) -3- (2-oxo-3-piperidinyl) -1-nitrosourea, characterized in that 1- (ß-chloroethyl) -3- (2-oxo-3-piperidinyl) urea is reacted with nitrous acid under anhydrous conditions. 5. Use of the nitrosourea derivatives mentioned in claim 1 or 2 as an active ingredient with anti-tumor and / or anti-leukemia effects and low toxicity for the manufacture of medicaments for the treatment of humans and animals. 6. Medicament which, in addition to carriers and customary additives, is a compound according to claim 1 or as active ingredient 2 contains. 409819/1190