Abstract
Full Text
Chemistry
N. E. GOLUBEVA, O. V. KIL’DISHEVA, and Academician I. L. KNUYANTS
CANCEROLYTIC PEPTIDES
By condensation of the N-formyl derivative of p-di-(β-chloroethyl)-amino-DL-phenylalanine (sarcolysine) with esters of various amino acids, cancerolytic dipeptides containing sarcolysine were obtained (Table 1).
\[ \ce{ \underset{\ce{ClCH2CH2}}{\overset{\ce{ClCH2CH2}}{>}N} - C6H4 - CH2CH(NHCHO)COOH + H2N-CH(R)-COOR' ->[\ce{C6H11NCNC6H11}] } \]
\[ \ce{ \underset{\ce{ClCH2CH2}}{\overset{\ce{ClCH2CH2}}{>}N} - C6H4 - CH2CH(NHCHO)CONH-CH(R)-COOR' + C6H11NH-CO-NHC6H11 } \]
Table 1
\[ \ce{ \underset{\ce{ClCH2CH2}}{\overset{\ce{ClCH2CH2}}{>}N} - C6H4 - CH2CH(NHCHO)-CONH-CH(R)-COOH } \qquad R' \]
| No. | R | R′ | Found, % C | Found, % H | Found, % Cl | Found, % N | Calculated, % C | Calculated, % H | Calculated, % Cl | Calculated, % N | m.p., °C | Yield, % | Recrystallization |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | \(\ce{-CH2-SCH2CH2Cl}\) | \(\ce{-COOC2H5}\) | 48.02 | 5.90 | — | 7.93 | 47.84 | 5.66 | — | 7.96 | 121–123 | 40 | From ethanol |
| 2 | \(\ce{-CH2CH2SCH3}\) | \(\ce{-COOC2H5}\) | 51.27 | 6.45 | 14.46 | 8.73 | 51.22 | 6.35 | 14.40 | 8.54 | 109–110 | 70 | From ethyl acetate–petroleum ether |
| 3 | \(\ce{-CH2CH2-SCH2C6H5}\) | \(\ce{-COOC2H5}\) | 57.03 | 6.15 | 12.29 | 7.30 | 57.03 | 6.21 | 12.47 | 7.39 | 95–96 | Quant. | From ethanol |
| 4 | \(\ce{H}\) | \(\ce{-COOC2H5}\) | 50.58 | 5.77 | 17.33 | 10.58 | 50.5 | 5.73 | 17.54 | 10.4 | 118–120 | » | From methanol |
On interaction of the ethyl ester of sarcolysine with p-di-(β-chloroethyl)-aminophenylacetic acid, the ethyl ester of p-di-(β-chloroethyl)-aminophenacetylsarcolysine was obtained:
\[ \ce{ \underset{\ce{ClCH2CH2}}{\overset{\ce{ClCH2CH2}}{>}N} - C6H4 - CH2-CH(NH-COCH2-C6H4-N(CH2CH2Cl)2)-COOC2H5 } \]
Next, p-di-(β-chloroethyl)-aminophenacetyl and γ-\[*p*-di-(β-chloroethyl)-amino\]-phenylbutyryl derivatives of various amino acids of the general formula were obtained:
\[ \ce{ \underset{\ce{ClCH2CH2}}{\overset{\ce{ClCH2CH2}}{>}N} - C6H4 - (CH2)_n CONH-CH(R')-COOH } \qquad \begin{gathered} R\\[-2pt] \end{gathered} \qquad n = 1,3 \quad \text{(Table 2)} \]
\[ \begin{gathered} (\mathrm{ClCH_2CH_2})_2\mathrm{N{-}C_6H_4{-}(CH_2)}_n\mathrm{CONH{-}CH(R'){-}COOH}\\ \hspace{8em} R \end{gathered} \]
Table 2
| \(n\) | R | R′ | Found, % C | Found, % H | Found, % Cl | Found, % N | Calculated, % C | Calculated, % H | Calculated, % Cl | Calculated, % N | M.p., °C | Yield, % | Recrystallization |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | —COOC₂H₅ | —CH₂C₆H₅ | 61.25 | 6.19 | 15.60 | 6.37 | 61.19 | 6.25 | 15.71 | 6.21 | 86—87 | Quant. | From ethanol |
| 1 | —COOC₂H₅ | —CH₂CH(CH₃)₂ | 57.42 | 7.27 | 16.37 | 6.9 | 57.54 | 7.24 | 16.98 | 6.71 | 81—82 | 47 | From ethanol |
| 1 | —COOCH₃ | H | 51.87 | 5.97 | 20.4 | — | 51.88 | 5.81 | 20.42 | — | 85—86 | 68 | From methanol |
| 1 | —COOCH₃ | —CH₂Cl | 48.71 | 5.58 | 27.20 | — | 48.60 | 5.32 | 26.92 | — | 113—114 | 79 | From methanol |
| 1 | —CH₂COOC₂H₅ | —C₆H₅ | 61.32 | 6.27 | 15.68 | 6.44 | 61.19 | 6.25 | 15.71 | 6.21 | 84—86 | 81 | From ethanol |
| 1 | —COOC₂H₅ | —CH₂CH₂—S—CH₃ | 52.18 | 6.39 | 15.90 | 6.37 | 52.41 | 6.48 | 16.29 | 6.44 | 46—49 | 91 | From ethanol |
| 1 | —COOC₂H₅ | —CH₂CH₂SCH₂C₆H₅ | 58.91 | 6.27 | 13.59 | — | 58.87 | 6.31 | 13.86 | — | 73—74 | 65 | From ethanol |
| 1 | —COOC₂H₅ | —CH(CH₃)₂ | 56.60 | 7.06 | 17.88 | 7.44 | 56.56 | 6.99 | 17.58 | 6.94 | 61—62 | 50 | From petroleum ether ethyl acetate |
| 1 | —COOC₂H₅ | —CH₂—S—CH₂CH₂Cl | 49.09 | 5.76 | — | 6.13 | 48.60 | 5.96 | — | 5.96 | 77—79 | 72 | From ethanol |
| 3 | —COOC₂H₅ | —CH₂C₆H₅ | 62.68 | 6.89 | 15.26 | 5.70 | 62.65 | 6.68 | 14.79 | 5.85 | 69—71 | 78 | From ethanol |
| 3 | —COOC₂H₅ | —CH—(CH₃)₂ | 58.47 | 7.70 | 15.75 | 6.37 | 58.46 | 7.48 | 16.44 | 6.5 | 62—63 | 92 | From ethanol |
| 3 | —CH₂COOC₂H₅ | —C₆H₅ | 62.53 | 6.73 | 14.75 | — | 62.65 | 6.68 | 14.79 | — | 84—86 | Quant. | From ethanol |
| 3 | —COOCH₃ | H | 54.31 | 6.45 | 19.09 | — | 54.40 | 6.40 | 18.92 | — | 84—85 | ” | From methanol |
By reacting \(p\)-di-(β-chloroethyl)aminophenylacetic and γ-\([p\)-di-(β-chloroethyl)amino]phenylbutyric acids with aniline in the presence of 1,3-dicyclohexylcarbodiimide, the corresponding anilides were obtained; however, in the reaction of these acids with 2-methyl-5-ethoxymethylene-6-aminopyrimidine, or of \(p\)-di-(β-chloroethyl)aminophenylacetic acid with cyclohexylamine, only the N-acyl derivatives of 1,3-dicyclohexylurea were isolated.
\[ (\mathrm{ClCH_2CH_2})_2\mathrm{N{-}C_6H_4{-}(CH_2)}_n\mathrm{CO{-}N(C_6H_{11}){-}CONH{-}C_6H_{11}}, \qquad n=1,3 \quad \text{(Table 3)} \]
\[ (\mathrm{ClCH_2CH_2})_2\mathrm{N{-}C_6H_4{-}(CH_2)}_n\mathrm{-CO-R} \]
Table 3
| \(n\) | R | Found, % C | Found, % H | Found, % Cl | Found, % N | Calculated, % C | Calculated, % H | Calculated, % Cl | Calculated, % N | M.p., °C | Yield, % | Recrystallization |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | —NH—C₆H₅ | 61.17 | 5.57 | — | 8.27 | 61.54 | 5.74 | — | 7.98 | 128—129 | 83 | From octane |
| 3 | —NH—C₆H₅ | 62.89 | 6.31 | 18.50 | 6.92 | 63.32 | 6.38 | 18.7 | 7.39 | 112—113 | Quant. | From octane |
| 1 | —N—CONH; substituents at N and NH: C₆H₁₁, C₆H₁₁ | 61.68 | 7.60 | 15.00 | — | 62.21 | 7.73 | 14.69 | — | 130—131 | 89 | From ethanol |
| 3 | —N—CONH; substituents at N and NH: C₆H₁₁, C₆H₁₁ | 63.36 | 8.08 | 13.42 | 8.16 | 63.51 | 8.09 | 13.89 | 8.23 | 128 | 51 | From ethanol |
Preliminary tests of the cancerolytic action of peptides containing sarcolysin were carried out at the Institute of Experimental Pathology and Cancer Therapy of the Academy of Medical Sciences of the USSR. It was found that sarcolysin peptides possess pronounced antitumor properties, close to those of sarcolysin, while at the same time they are nontoxic and have a high selectivity of action on certain tumors \((^{1})\).
General method for obtaining peptides containing sarcolysin
To a suspension of 1 mole of N-formylsarcolysin in chloroform, equimolecular amounts of 1,3-dicyclohexylcarbodi-
imide and the corresponding amino acid ester. The mixture was shaken vigorously and left at room temperature overnight. The precipitated 1,3-dicyclohexylurea was filtered off; after distillation of the chloroform, the residue was mixed with a small amount of alcohol* and left to crystallize in a refrigerator. The precipitated dipeptide was filtered off (see Table 1).
Ethyl ester of n-di-(β-chloroethyl)-aminophenacetylsarcolysine
Equimolecular amounts of ethyl sarcolysinate, n-di-(β-chloroethyl)-aminophenylacetic acid, and 1,3-dicyclohexylcarbodiimide were left in chloroform solution overnight. The precipitated 1,3-dicyclohexylurea was filtered off, the solvent was distilled off in vacuo, the residue was dissolved in ethyl acetate, petroleum ether was added until turbidity appeared, and the mixture was left to crystallize in a refrigerator.
Ethyl ester of n-di-(β-chloroethyl)-aminophenacetylsarcolysine: colorless crystals, m.p. 92–94° (from ethanol). Yield 95%.
\[ \begin{aligned} &\text{Found, \%: } \mathrm{C}\ 54.81;\quad \mathrm{H}\ 5.93;\quad \mathrm{Cl}\ 24.09\\ &\mathrm{C}_{27}\mathrm{H}_{35}\mathrm{Cl}_{4}\mathrm{N}_{3}\mathrm{O}_{3}.\ \text{Calculated, \%: } \mathrm{C}\ 54.82;\quad \mathrm{H}\ 5.92;\quad \mathrm{Cl}\ 24.03. \end{aligned} \]
General method for preparing esters of N-n-di-(β-chloroethyl)-aminophenacetyl- and N-γ-[n-di-(β-chloroethyl)-amino]-phenylbutyrylamino acids
A mixture of equimolecular amounts of the amino acid ester of n-di-(β-chloroethyl)-aminophenylacetic (or γ-[n-di-(β-chloroethyl)-amino]-phenylbutyric) acid and 1,3-dicyclohexylcarbodiimide in chloroform was left at room temperature overnight. The precipitated 1,3-dicyclohexylurea was filtered off, the solvent was distilled off, and the residue was mixed with a small amount of alcohol* and left to crystallize in a refrigerator (see Table 2).
Anilides of n-di-(β-chloroethyl)-aminophenylacetic and γ-[n-di-(β-chloroethyl)-amino]-phenylbutyric acids
A mixture of equimolecular amounts of aniline, the above-mentioned acids, and 1,3-dicyclohexylcarbodiimide was left in chloroform overnight. The precipitated 1,3-dicyclohexylurea was filtered off and thoroughly washed with chloroform. The solvent was distilled off in vacuo. The anilides obtained were recrystallized from alcohol (see Table 3).
Ethyl ester of s-β-chloroethylcysteine
3 g of chloroethylcysteine hydrochloride was boiled in 50 ml of \(\mathrm{C}_{2}\mathrm{H}_{5}\mathrm{OH}\), saturated with HCl, passing a dry stream of HCl for 2 h. The solvent was distilled off to dryness. The hydrochloride of ethyl ester of chloroethylcysteine was obtained as colorless needles, m.p. 108–110°. Yield 81%.
\[ \begin{aligned} &\text{Found, \%: } \mathrm{C}\ 33.83;\quad \mathrm{H}\ 6.01\\ &\mathrm{C}_{7}\mathrm{H}_{15}\mathrm{Cl}_{2}\mathrm{NO}_{2}\mathrm{S}.\ \text{Calculated, \%: } \mathrm{C}\ 33.88;\quad \mathrm{H}\ 6.09 \end{aligned} \]
* In preparing ethyl ester of N-formylsarcolysylmethionine, the residue was dissolved in ethyl acetate and petroleum ether was added until turbidity appeared.
* Ethyl ester of s-β-chloroethylcysteine was used in the form of an oil, which was isolated from the hydrochloride of ethyl ester of s*-β-chloroethylcysteine with sodium bicarbonate.
* In the case of condensation of γ-[n-di-(β-chloroethyl)-amino]-phenylbutyric acid with esters of phenylalanine or valine, the residue was dissolved in ethyl acetate and petroleum ether was added until turbidity appeared.
N-\(n\)-di-(β-chloroethyl)-aminophenacetyl and N-γ-\([(p\)-di-(β-chloroethyl)-amino]-phenylbutyryldicyclohexylurea
A mixture of equimolecular amounts of \(n\)-di-(β-chloroethyl)-aminophenylacetic or γ-\([n\)-di-(β-chloroethyl)-amino]-phenylbutyric acid, 1,3-dicyclohexylcarbodiimide, and 2-methyl-5-ethoxymethylene-6-aminopyrimidine was left in chloroform solution overnight. The solvent was distilled off; the residue was mixed with ethyl alcohol and allowed to crystallize (see Table 3).
Institute of Organoelement Compounds
Academy of Sciences of the USSR
Received
10 XII 1957
REFERENCES CITED
- L. F. Larionov, Z. P. Sof’ina, DAN, 114, 1070 (1957).