DD209192A5 - METHOD OF PREPARING N- (N '- (2-CHLOROETHYL) -N'-NITROSO-CARBAMOYL) -OLIGOPEPTIDE ESTERS AND -AMIDES - Google Patents

Abstract

Compounds having the formula: <IMAGE> wherein: <IMAGE>R4 = -OCH3, -NH2, NH-R4, R4 being an oligopeptidic residue of which the terminal carboxyl group is blocked as a methyl ester or amide group. They are prepared by reacting the corresponding aminoacids or oligopeptides with an ester or an amide of a second aminoacid or of a second oligopeptide, in presence of an appropriate basic peptidic catalyst.

Description

Berlin, the 11th of '83

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Process for the preparation of ltf-jJJ ^f - (2-chloro-3-thyl; L) -iJ'-nitroso-carbamoylj-oligopeptide esters and amides

Field of application of the invention

The invention relates to a process for the preparation of JT-JjP- (2-chloroethyl) -I-nitroso-carbamoylJ-oligopeptide esters and amides.

The compounds according to the invention are used as medicaments, in particular for the treatment of cancer,

Characteristic: the known technical solutions

Among the alkylating agents, a new class of substances has been introduced into cancer chemotherapy in recent years, namely, the uritro-osmotic agents, eg, 1,3-bis (2-chloroethyl) -1-nitrosourea (BGlU), 1- (2 Chloroethyl) -3-cyclohex-1-1-nitrosourea (CCSU) and 1- (2-chloroethyl) -3- (4-methylcyclohexyl) -1-nitrosoharnate (MeGCOT) (Proceeding of the 7th New Drug Seminar on the Nitro3oureas , (Washington, D "C, Dec." 15-16, 1975), Cancer Treat, Rep., Op. Cit., 645-811 (1976).) In order to obtain substances with better therapeutic index, there has recently been a number new nitrosoureas with various sub-substituents, see B., sugar derivatives, synthesized and experimentally tested (Anderson et al., Cancer Res., 3, 761 (1976), Kayat et al., Cancer Chemother.Pharmacol., 3, 217 (1979 ).

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Destination of the ship

The aim of the invention is to provide new compounds with better therapeutic index which are suitable for cancer chemotherapy.

Explanation of the essence of the invention;

It is the object of the invention to find new compounds having the desired properties and processes for their preparation.

In contrast to the previously synthesized substances, in the present Pail are oligopeptide esters which carry on the amino group an iT- (2-Ghloräth ^) T, nitroso carbamoylgruppe. As a starting material for the synthesis of Ii] ks' - (2-chloroethyl) -I' nitrosoj-carbamoyl-oligopeptidestern serve lni ^f - (2-ChloräthyDlJ'-nitroso] carbamoyl or -oligopeptide, in turn, conveniently prepared by reaction of.! - (2-Chloroethyl) -Nn.itrosocarbamoylazide with amino acids or oligopeptides are accessible under the influence of known per se peptide catalysts such as, for example, Dicyclohesylcarbodiimide DCC) react the mentioned lel ^l - (2-chloroethyl) U'-nitrosoJcarbamoylaminosäuren or -oligopeptide amides, so that they with. the ester or amide of a second amino acid or a second oligopeptide form a peptide bond. The reaction can be schematically represented as follows:

i i u

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GOR ₀ COR.

t ^ά ι 4

(ID ff £ i

HO R ₁ R

(D

COR ₄

COHH-CH

CI-CH ₀ -CH ₀ -H-Co-HH-CH E-

HO R ₁

(III) R ₁ = R ₃ = -H, CH ₃ , -CH (CH ₃ ) ₂ , -CH ₂ -CH (CH ₃ ) ₂ , -CH (CH ₃ ) C ₂ H ₅

_s -CH ₂ OH, -CH (OH) CH ₃ , -CH ₂ -CH ₂ -S-CH ₃ , 1 j-CH ₀

R ₂ = OH or · -HH-R ', where RA represents a peptide group with a free carboxyl group which is peptidically bound via the terminal amino group,

Examples of R ':

-Gl ^ cyl-glycine

Glycyl-glycyl and peptides of other amino acids up to the tripeptide, as well as mixed oligopeptides such as:

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Glycyl-Alanine Alanyl-Leacin Leucyl-Alanine Glycyl-Tyrosine

 Leacyl-glycyl-tyrosine Leucyl-glycyl-proline

R ₄ = OCH ₃ , Mi ₂ , -HR », where R | Alkyl or aryl or an oligopeptide residue which is peptide-bonded via the terminal amino group and whose terminal carboxyl group is in turn present as methyl ester or amide.

Examples of Ri .:

-GIycin-Methylester

-GIycy1-Glycy1-MethyIeater

-GIycy1-Glycy1-GlycyImethy1ester

Peptides of other amino acids to tripeptide as well as mixed oligopeptides such as:

Glycyl-alanine Methylester

Leacyl-glycyl-proline

Leacyl-alinin amide

GIycyl tyrosine amide

The reaction of the starting material I with the ester or amide of another amino acid or of an oligopeptide II, which may be the same or different from the first amino acid or the first oligopeptide, is preferably carried out under cooling (about ⁰ ° C.), but also without problems room temperature. For example, ImId ii- (l- ^t - (2-chloroethyl) -LT ^! -NitrosocarbamoyD-amino acid and 1 ΩM amino acid methyl ester are dissolved together in 20 ml of dichloromethane In this solution is added with stirring 1.1 raM (230 mg) dicyclohexylcarbodiimide, The mixture was dissolved in 10 ml of dichloromethane and slowly added dropwise

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The mixture is then stirred for a further hour and left to bubble over at room temperature. The filtrate is filtered off under reduced pressure after filtering off Itfjli'-dicyclohexylurea. The crude product is obtained, which is purified by column chromatography on silica gel using ether / dichloromethane of different mixing ratios. After slution, the main fractions are combined and concentrated under vacuum. The residue is taken up in dichloromethane, if necessary admixed with n-pentane, and crystallized in a refrigerator. »

Particularly valuable for cancer chemotherapy are the following compounds of the invention:

Ιί- £ ϊΡ- (2-chloroethyl) -! ' nitroso) -carbamoyl-glycyl-L-valine methyl ester

IJ- [JET '- (2-chloroethyl) -35T-i-nitroso) -carbamoylj-L-valyl-L-valine methyl ester)

E-lss ¹ - (2-chloroethyl-3Tl) -H '-nitroso) -carbamoylj-L-valvl-L-methionine methyl radicals rs

ÜT-yyj »- (2-chloroethyl) -iI '-nitroso) -carbamoylJ-L-isoleucy 1-L-leucine methyl ester;

SJ] JJ ₁ '- (2-chloroethyl) -BP -nitroso) -carbamoylJ-L-phenylalanyl

L-valine methyl ester; ,

H- | jS »- (2-chloroethyl) -1ϊ'-nitroso) -carbamoy IJ-L-phenylalanyl-1-leucine methyl ester;

H-rsr »- (2-chloroethyl) -Ii-nitroso) -carbamoyl-L-phenylalanyl-L-phenylalanine methyl ester;

Ii-jjp (2-chloroethyl) -li- ^f- aitroso) -carbamoylj-L-phenylalanyl-L-methionine methyl ester;

£ j- [igi- (2-chloroethyl) -I ^! nitroso) carbamoyl Ij-L-methionyl-L-phenylalanine methyl ester.

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Ausfuhrangabeispiel

The following examples illustrate the invention:

example 1

Preparation of U-jjf '- (2-chloroateJayl) -li'-nitroso-carbaioioyl] -glycyl-L-valine methyl ester (general formula III) R ₁ a -H ₅ R ₂ = - CH (CH ₃ ) ₂

Tj-jjft _ (2-chloroethyl) -srn-nitroso-carbamoyl) -glyloine (1mM) and L-valine methyl ester (1mM) are dissolved together in 20mL of dichloromethane. Dicyclohexylcarbodiimide (DCG, 1.1 ml), dissolved in 10 ml of dichloromethane, is added dropwise with stirring to this solution. The reaction mixture is then stirred for a further hour and allowed to stand over hackle at room temperature, filtered off by filtration and '-Dicyclo-hexyl-urea, the filtrate is concentrated under vacuum. The crude product is purified by column chromatography (silica gel, eluted with ether / di-chloroethane 1: 10). From the main fractions, the pure product after removal of solvent, in vacuo and by recrystallization from dichloromethane / n-pentane is obtained * pale yellow JSadel, yield; ca, 70 ^ fI ¹ · '97 to 98 ⁰ G (Zers,); Elemental analysis: Ber.i G 4O, 94; H 5.93; U 17,36 »Gef .: C 41,09; H 6,00; Έ 17j16, The H-HMR spectroscopic study revealed characteristic peaks at if = 0.95 Ppm (m, -G (GH ^) _p );

D = 2.20 ppm (m, -CE =); cf = 3.50 ppm (t, Cl-CH ₂ -);

6 = 3.75 ppm (s, -OCH ₃ ); S = 4.20 ppm (-Ji-CH ₂ -CO- and -CH ₂ -I-UO); cf = 4.60 ppm (m, -N-CH-C-); S = 6.70 ppm. (d, -1Ώϊ-) 5 (5 = 7.70 ppm (t, -NH-), The mass spectrum shows no signal for a molecular ion, but 'characteristic fragments in M-I07 and M-108, the cleavage of Cl (GH ") 2" M) or · Cl (GH ₂ ) ₂ -N = H0H.

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Example 2

Preparation of N, N'-chloroethyl-1H'-nitro-carbamovl-1-valvl-L-valine methyl ester (general formula III)

I_Qj »_ (2-Cliloräthyl) -I \ f ^l -nitroso-carbaiiioyj-L-valine (1 mM) and L-valine methyl ester (1 mM) are treated analogously as described in example. 1 Bas product crystallized from dichloromethane / n-pentane as pale yellow needles, yield 65 %, mp .125 to 126 ⁰ C (dec.) I Slementaranalyse; 3 .; G 46.09; H. 6.91; N 15,365 Found: G 46,31S H 7,10, U 15,23. The ¹ H-HMR spectroscopic analysis revealed the following characteristic peaks: cf = 0.9-1.1 ppm (m, -G (CHo) ₂ ) i & = 2.20 ppm (m, -GH =) ; € = 3.50 ppm (t, Cl-CH ₂ -); 6 = 3.75 ppm (s, -OGH ^); O = 4.20 ppm (t, -GH ₂ -Ui-IJO); 6 = 4.50 ppm (m, -ii-CH-GO); ί = 6.60 ppm (d, -IiH-); cT = 7.50 ppm (d, -JIH-). The mass spectrum shows a molecular peak at 364 and peaks at m / e = 258 and 257 (M-I07 and M-108). The last two peaks are formed by cleavage of Gl-CH ₂ -GH ₂ -IJ-UO or Gl- CHg-CH ^ U-iJOH, either by simple cleavage of the ST-C bond or by a Mclafferty rearrangement,

Example 3

Preparation of I-jjf ¹ - (2-Chloräthvl) -I ^l -nitroso-carbamovlJ-L-valvl-L-methioninmethvlester (general Pormel III) R ₁ = -CH (CH _{3) 2;} R ₂ = S-CH ₂ -GH ₂ -SCH ₃ )

Example ¹ - (2-Chloroethyl J-Ii ¹ -nitroso-carbasioyl) -L-Yalin (1 sM) and L-methionine methyl ester (TiaM) are prepared analogously as described under

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Example 1 treated. Yellowish needles (dichloromethane / n-pentane), yield about 70 % ,? .81 to 82 ⁰ G (Zers.). Elemental Analysis: Calc. G 42,37i H 6,35; Ii 14,12; Found: C 42.65; H 6,50; M 14,14. The H-SMR spectroscopic analysis revealed the following characteristic peaks: S = 1.00 ppm (in, -C (OE.,) ₂ ) i Cf = 2.10 ppm (-GH ₂ -S-GH ₃ J -GH =); 6 = 2.55 ppm (t, -GH ₂ -); α "= 3.50 ppm (t, Cl-GH ₂ -)"<r = 3.80 ppm (s, -OCH ₃ ), 6 = 4.20 ppm (-CH ₂ -H-IiO), 6 = 4.40 ppm (m, -2J-CH-CO); cf = 4.80 ppm (m, -2T-GH-GO) J (f = 6.80 ppm (d, SH-); <f = 7.50 ppm (d, -SH-), Bas mass spectrum shows a molecule peak at 396 and peaks at m / e = 289 and 288 (m-107 and M-108),

Example 4 .

Preparation of H-1LT ^} - (2-chloroethyl) -Ii'-nitroso-carbamoyl-L-isoleuc-1L-leucine methyl ester (general formula III) R ₁ = -GH (CH ₃ ) CH ₂ CH ₃ ; E ₂ = -CE ₂ -GH (CH ₃ ) ₂ )

jj-jjf '- (2-chloroethyl) H ⁱ -nitroso-carbamoyl] -L-iso-leucine (1 mM) and L-leucine methyl ester (1 mM) are treated analogously as in Example 1. Light yellow needles (dichloromethane / n-pentane), yield about 60 %, I ¹ .. 128 to 129 ⁰ C (cerium). Elemental analysis: calc .: G, 48.91; H 7,44; H 14,26; Found .; G 49.03; H 49.03; H 7.70; Ή 13.96 "The ^ H-JSHR spectroscopic examination revealed the following peaks:. O = 1.00 ppm (-GH ₃ ); (S = 1.25-2.00 ppm (-GH =; -GH ₂ -); cf = 3.50 ppm (t, Gl-GH ₂ -); (Γ = 3.80 ppm (s, -OGH ₃₎ CF = 4.20 ppm (t, GH ₂ -S-IiO). 4.40 ppm (m, -N-CH-CO) rCf = tf = 4.70 ppm (m, -F-CH D = 6.30 ppm Cd, -BH-) 5 6 = 7.45 ppm (d, -HH-). The mass spectrum shows a molecule peak at 392 and peaks of ni / e = 286 , 285 (M-107, M-108),

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Example 5

Preparation of LiI ^T - (2-chloroethyl) -! F-nitroso-carbamoyl] -L-phenylalanyl-L-valine methyl ester (III, R ₁ = -CH ₂ -CgHc; R ₂ = -CH (CH ₃ ) ₂ )

N- (IP- (2-chloroethyl) -F'-nitroso) -carbamoyl-L-phenylalanine (1 mM) and L-valine methyl ester (1 mM) are treated analogously as in Example 1. Yellow ladein (dichloromethane / n-pentane), yield about 60 %, I ¹ .107 to 108 ⁰ C. Elemental analysis; Ber. G 52.36; H 6,10; SF 13.57; Gef.:. C 52.54; H 6.21; IT 13.28 ·

The H-iSMS spectroscopic examination gave the following peaks: <5 = 0.85 ppm (m, -C (CH-) ") j (T = 2.10 ppm (m, -CH =); 6 = 3 , 15 ppni (d, -CH ₂ -ph); C = 3.40 ppm (t, Cl-CH ₂ -); S = 3.70 ppm (s, -OCH ₃ ); cf = 4.10 ppm ( t, -CH ₂ -S-UO); cf = 4.45 and 4.80 ppm (m, -I-CH-CO), 6 = 6.35 ppm (d, -SE-), ^ = .7.25 ppm (-CgH ₅ -); cf = 7.55 ppm (d, -MH-). The mass of the supernatant shows a molecular peak at 412 and peaks at m / e = 305, 304 (FIG. M-107, M-108),

Example 6

Preparation of I- [li ¹ - (2-chloroethyl) -j5'-nitroso-carbamoyl] -L-phenylalanyl-L-leacine methyl ester (general formula III) R ₁ A-CH ₂ -CgH ₅ JR ₂ = - CH ₂ -CH (CH ₃ ) ₂ )

U £ ii ^l - (2-chloroethyl) ^-I-t -nitroso carbamoyl3-L-alanine piieny (IMM) and L-leucine methyl ester (1 mM) are treated analogously as described in .Example. 1 Pale yellow saddles (dichloroethane / η-pentane), yield about 60 fi> ₉ ? _D * 96 to 96.5 ⁰ C. Eleraentaranalyse: Ber ": C 53.46; H 6,38; U 13:12; Found .:

L ^ ü j

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C 53,29; Ξ 6,34 ίΤ 12,92.

H-MR spectroscopic analysis revealed characteristic reactions; cf - 0.90ppm (-C (CH ₂ ) ₂ ); S = 1.50 ppm (m, -GH ₂ -CH =); <f = 3.15 ppm (d, -CH ₂ -ph); (f = 3.45ppm (t, Cl-CH ₂ -); 6 = 3.75ppm (s, -OCH ₃ ); <f = 4.10ppm (t, -CH ₂ -H-NO) ;

Cf = 4.55 ppm (m, -IT-CH-CO); d = 4.80 ppm (m, -N-GH-CO); £ = 6.20 ppm (d, -HH-) j cf = 7.30 ppm (-CgH ₅ ); cf = 7.50 ppm (d, -HH-).

The mass spectrum shows molecular peaks at 426 and peaks at m / e = 319, 318 (M-107, M-108).

Example 7

Preparation of H-IJI'-X-C-chloroethyl-N'-nitroso-carbamoyl-1-phenylalanyl-L-phenylalanine methyl ester (general formula III) R ₁ = R ₂ = -CH ₂ -C ₆ H ₅ ) ,

H-Qs "- (2-chloroethyl) -! '- nitroso-carbamoyl] -! - phenylalanine (1 mM) and L-phenylalanine methyl ester (1 eq) are analogous. treated as in Example 1. Pale yellow needles (Dichlornaethan / xL-pentane), yield 60%, P .116 to 117 ⁰ G. Blementaranalyse: Calc .: 57.33 G »H, 5.47 ;: IT 12.16; Gef .: 'G 57,22; H 5,23; U 12,12.

1 - '

H-NMR spectroscopic examination gave the following peaks: = 3.10 ppm (CH ₂ -ph); S = 3.40 ppm (t, Gl-CH ₂ -);

<$ = 3.70 ppm- (s, -OCH ₃ ) TcT = 4.10 ppm (t, -CH ₂ -I-IO);

6 = 4.75 ppm (-U-CH-CO); </ = 6.10 ppm (d, -3H -); (T-7.20 ppm (-CgH ₅ ); (f = 7.40 ppm (d, - HH-).

The mass spectrum shows molecular peaks at 460 and peaks at m / e = 353, 352-. (m-107-, M-108) ·

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Example 8

Herateilung of H- [l ^t - ^(2-t Chloi a-thyl) -lT'-nitroso-carbanio7l] -L-phenylalanyl-L-ciethioninietholester (general Pormel III)

B-jjj '- (2-Chloräth ^ l) -I3' ^f -nitro3o-carbanioylJ -L-phenylalanine (1 mH) and L-methionine (1 mM) are treated analogously as described in example. 1 Pale yellow ladein (Di chlorine than / n-pentane), yield ca, 70%, Ϊ .74 to 75 ⁰ G. Elemental analysis: Calc., C, 48.59; H 5.66; Έ 12.59; Found .:

G 48,69i H 5,67 »U 12,41 ·

The H-HMR spectroscopic examination revealed the following

characteristic peaks at: tf = 2.05 ppm (-CH ₂ -S-CH-); 6 = 2.45 ppm (-CH-, -); 6 '= 3.20 ppm (d, -CH.ph); 6 '= 3.45

C. C.

ppm (t, Cl -CH ₂ -); cf = 3.75 ppm (a, -OCH ₃ ); cT- 4.10 ppm (t, -GH ₂ -IT-IiO); <S = 4.65 ppm (m, -ST-CH-CO); <f = 4.30. ppm (m, -U-CH-CO); if = 6.50 ppm (d, -SiH-); (f * 7.25 ppm UCgH ₅ );

cf = 7 _v 5O ppm (d, -IaH-)

 The mass spectrum shows a molecular peak at 444 and peaks at m / e = 337.336 (M-107, M-108).

Example 9

Preparation of τοη B - [JT ^f - (2-chloroethyl) -ii'-nitroso-carbamoyl-L-aiethioyl-L-phenylalanine, ethyl ester (general formula III) R 1

H-1-yl (2-chloroethyl) -ii'-nitroso-carbamo: / 1-L-inethionine (1 mM) and L-phenylalanine methyl ester (1 mM) are treated analogously as in Example 1 * pale yellow aliquots (dichloromethane / n Pentane), yield ca, 70 %, F, 70 to 71 ⁰ C,

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Elemental analysis: 3rd: C 48.59; H 5.66; N 12.59; Found: C, 48.64; H 5.68; Ή 12.36,

The H-HMR spectroscopic examination gave the following peaks: <Ta 2.05-2.20 ppm (-GH ₂ -S-CH ₃ ); & = 2.65 ppm (t, -GE ₂ -); <f = 3.10 ppm (d, -CHg-pYa); <f = 3.45 ppm (t, Gl-GH ₂ -); Cf = 3.75 ppm (3, "- OGH ₃ ); d = 4.10 ppm (t, -CH ₂ -U-IO); -.rf = 4.70-4.90 ppm (m, -BT (F = 6.80 ppm (d, -SE-); <f = 7.20 ppm (m, -G ₆ H ₅ ); cf = 7.65 ppm (d, -IiH-) The Maasenspek fcrum shows a molecular peak at 444 and peaks at m / e = 337, 336 (M-107, M-198).

Claims (3)

Srfindungsanspruch 1. A process for the preparation of iJ- [jM ^f - (2-chloroethyl) ii'-nitroso-carbanioyl] -oligopeptide esters and amides of the general Pormel 00R ₄ COM-GH CI-GH ₀ -CH ₀ -N-Go-IH-GH L (III) and IiO. R ₁ wherein , = E ₃ = »- H, CH ₃ - -CH (CH ₃ ) ₂ , -CH ₂ -CH (GH ₃ ) ₂ , -G ₃ H ₅ -CH (CH ₃ XO ₂ H ₅ , -CH ₂ - (0 ), -GH ₂ OH, -GH (OH) CH _3, -CH ₂ -CH ₂ -S-GH _3, -CH ₀ -GH ₀ -C -UH ₀ -CH ₀ -π- 0 ^] H s -OCH ₃ , - & S ₂ "M-R ', wherein E» represents an oligopeptyl radical whose; terminal carboxyl group is in turn closed as a methyl ester or ainide group, characterized in that N-1-L '- (2-chloroethyl) -H ¹ -nitro so car bamoylj amino acids or oligopeptides with an ester or amide of a second amino acid or oligopeptide, which may be the same or different, under the action of a suitable basic peptide catalyst in the cold, su ^~ (3-chloro, ethlyI) -I'-nitroso-carbaoyl oligopeptide esters or amides are reacted y y 02/11/1983 AP C 07 D / 243 011/0 (61 384/11) 2. The method according to item 1, characterized in that the used as starting material
Η-pi ¹ (2-chloro-or- ¹ -yl) -IT ¹ -nitroso-carbamoyl] amino acids or oligopeptides of H- (2-chloroethyl) -7-nitroso-carbamoylazide and amino acid or
Oligopeptid be prepared in a conventional manner in a suitable solvent, in particular Dichloride than and this reaction solution is used without isolation for the reaction with the amino acid ester or amide or corresponding oligopeptide 3 · Procedure according to item 1 or 2, marked
in that as a basic Petidkatalvsator
Dicyclohexylcarbodiimide is used,