NL7905098A - NEW DAUNORUBICIN DERIVATIVES, PROCESS FOR THEIR PREPARATION AND PHARMACEUTICAL PREPARATIONS CONTAINING THEM. - Google Patents

Description

. «; for 7859 Rhone-Poulenc Industries I Paris, France.

: Novel daunorubicin derivatives, process for their preparation and pharmaceutical preparations containing them.

The invention relates to new daunorubitin derivatives of the general formula T of the formula sheet, optionally their addition salts with acids, their preparation and preparations containing them.

5,. In general formula 1, the symbol Rj represents a radical of the general formula 2 of the formula sheet, "in which"! the symbols X ,, representing an atom of oxygen or sulfur ,, iden-! tics. jrr and the symbols are alicyl- * phenyl or phenyl substituted in the p-position by a methyl, methoxyl or methylthio radical or represent together an alkylene radical with 2-b carbon atoms. or the symbols X are different and the symbols R 4 together form an alkylene radical with 2 - b carbon atoms, and it; symbol R ^. represents a radical of the general formula 3 of. the formula sheet, wherein R 1 represents a hydrogen atom or an alkyl, phenyl or benzyl radical.

Furthermore, it is to be understood that the above-mentioned alkyl radicals and parts, straight or branched, contain 1 to 1 carbon atoms and the compounds of the general formula 1 are optionally derived from the forms D, L and DL of the amino acid of the general formula Ij, wherein is as described above.

According to the invention, the compounds of the general formula 1 can be prepared by the action of a reactive derivative of an amino acid of the general formula k, the amine functions of which are pre-protected on a daunorubicin derivative of the general formula 5, wherein R 1 is as described above or on one of its salts.

The amine function of the amic acid of the general formula 11 can be protected by any method known per se for blocking the amino radical of an amino acid and. '; 3-. ; "-". ·., * -. - '·' '' ~ 2 —Λ '* * .- ’r *; of which, the lock and unlock conditions. tolerate, with the stability of the molecule.

The acid function of the amino acid of the general formula k may be activated either in the form of mixed arthydric acid or in the form of reactive ester.

1) The acid function of the amino acid of the general. formula 4 may be activated by forming a mixed anhydride 1 by reacting the amino acid with e.g. an ethyl isobutyl or sec-butyl chloroformate.

10 'If one. the amino acid of the formula in the form.

| of a mixed anhydride (which may have been prepared in situ), the amine function is protected for all by a radical [(biphenylyl -: - ^) - 2. propyl-2] oxycarbonyl which can be introduced according to the method described by Sieber and B. Iselin, Helv. Chim. Acta. 51, Ji 15, 622: (1968), and which is easily removed in dilute acid medium, | especially in the presence of hydrochloric acid j

De-condensation of the mixed anhydride of the product of formula k on the daunorubicin derivative. of the general formula 5 then takes place: in an anhydrous medium in an organic solvent, such as methylene chloride or tetrahydrofuran, in the presence of a nitrogen base, such as, triethyl amine or U-methyl morpholine at a temperature between -20 and 0 ° C. Preferably one works under nitrogen.

The removal of the. protecting group of the amine function. takes place in the presence of. chlorine what there is hydrochloric acid in - an organic solvent, such as methylene chloride at a temperature between b and 25 ° C.

2) The acid function of the amino acid of the general formula U may also be activated by esterification with hydroxy-3 xyl compounds, such as N-hydroxy succinimide. This activated ester can be prepared in situ.

When operating according to this variant, the amine function of the amino acid of the general formula (U) can be protected by a group, such as the trityl radical, which can be removed in dilute acid medium.

\ 7 90 5 0 98 i. Among these conditions, the condensation reaction of the activated ester with faet produces product of the general formula 5 in an organic solvent such as ethyl acetate or dimethyl formamide in the presence of a carbodiimide such as dicyclohexyl -5 carbodiimide at a temperature between -15 and + 25 ° C, optionally at. presence of an organic base, such as trietfaylamine.

The products of. the general formula 5 can be obtained according to the method described in Faet Belgian Patent Fee 8½. 219.

The new compounds of the general formula 1 can optionally be purified, by physical methods (such as by crystallization, or chromatography) or chemically (such as by salting, crystallization and subsequently decomposition).

The new compounds according to the invention can optionally be converted into acid addition salts with acids.

The salts can be obtained by the action of the new compounds on acids in suitable solvents. As organic solvents used, e.g. alcohols, ethers, ketones or chlorinated solvents, the salt formed precipitates after optionally 20. concentrate the solution down and separate by filtration or decanting.

The new derivatives of the daunorubicin of the general formula 1 as well as their salts have interesting anti-tumor properties combined with a low toxicity.

They have been shown to be particularly active on mouse-grafted tumors, especially at doses of 0.20-20 mg / kg intraperitoneally on leukemia L 1210.

Their maximum tolerated dose is determined, in the mouse it is 0.5 - 30 mg / kg intraperitoneal.

Of particular interest are the compounds of the general formula 1, wherein a radical is of the general formula 2, in which the symbols X represent oxygen and the symbols R 1 alkyl radicals are either the symbols X, which may or may not be the same, represent oxygen or sulfur and the symbols Rg together form an alkylene-35 radical with 2-carbon atoms and Rg is a leucyl-radical.

...... .... .. ..

90 Ι 7905098 Q ..

- 4 - / In particular - as special active compounds; Named #: (K (L-leucyl) -diethoxya.cetoxy-t4-daunorubicin and 11- (L-leucyl) - (ditbiolan-T, 3-yl-2) carbonyloxy-1,4-daunorubicin.

For therapeutic application, the. novel daunorubicin derivatives of the invention are used either in free or in the. form. of pharmaceutically acceptable salts, i.e. non-toxic at the use doses i

As examples of pharmaceutically acceptable salts, mention may be made of salts of mineral acids (such as chlorohydrates, sulfates, nitrates and phosphates) or organic acids (such as acetates, propionates, succinates, benzoates, maleates, maleates). ,.

tartrates ,. theophylline acetates, salicylates ,. phenol phosphates, - | . . : methylene bis-B-oxynaphthoates). "; ; The invention is further elucidated by means of the examples below. ; Example Γ!

In a pre-dried three-necked round-bottom flask, it is dissolved under nitrogen atmosphere-T, 66 g of LNi [(bif enylyl-4) -2-propyl-2-] 3 3. oxycarbonylleucine in 100 cm of tetrahydrofuran and adds 2.9 cm of triethylamine, stored on potassium carbonate lozenges. It is cooled to -20 ° C and. once added 2.48 cm 2 of isobutyl chloroformate. It is stirred at: -20 ° C for 30 minutes. It is added all at once while maintaining the temperature of. the reaction mixture at -20 ° C a solution of. 12.3 g of diethoxyacetoxy-1,4-daunorubicin chlorohydrate 25 in 200 ml of dry methylene chloride, containing 2.43 cm of triethylamine, stored on potassium carbonate lozenges. It is stirred at -20 ° C for 15 minutes.

3 ..

1000 cm of ice water is added, decanted and the organic phase is washed three times with 200 cm of water. The organic phase is dried over sodium sulfate. It is filtered and concentrated to dryness under reduced pressure 300 (20 mm Hg) at 30 ° C. The residue is dissolved in the minimal amount of methylene chloride and this solution is poured onto a column (diameter 5 cm, height 60 cm) containing 450 g of silica gel in chloroform

O

contains. Elution is carried out successively with 4000 ml of methylene chloride,

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containing 10% ethyl acetate, then with 5,100 cm3 of methylene chloride, containing 25% by volume ethyl acetate to recover an eluate with fractions of Tan 300 cm @ 790 5 0 9 8 3 .

The fractions 1 ^ -30 are combined, concentrated to dryness under reduced pressure (20 mm mercury) at 30 ° C and then dried under reduced pressure (1 mm mercury) at 20 ° C.

11.3 g of H- [N - {[(biphenylyl- * 0-2 propyl-2] oxyearbonyl} L leucyl] diethoxyacetoxy-1-daunorubicin in the form of an amorphous red solid are thus obtained dust ;.

[α] 25 - +126 - 16 ° (c = 0.182, chloroform).

11.3 g of [(biphenylyl- * 0-2 propyl-2] - are dissolved

O

Oxycarbonyl} L. leucyl] diethoxyac etoxy-1 * (- daunorubicin on 130 ml of methylene chloride. The mixture is cooled to + k ° Q in a .3 bath, of ice and water. It is stirred once 25.7 cm: of a Q 77H dry hydrogen chloride solution in dioxane Ha 10 minutes.

3: 300 ml of ethyl acetate are added and. 500 cm3 ethyl ether. The red precipitate obtained is filtered, washed with ethyl ether and dried under reduced pressure (1 mm mercury) at 20 ° C. It. red powder is suspended in 100 cm ethyl acetate. It is stirred for 15 minutes and the mixture is filtered. 3 mourns. The red solid obtained is washed three times with 100 cm ethyl acetate and dried under reduced pressure (1 mn mercury) at 0 ° C. 6.62 g of IT- (L-leucyl) diethoxyacetoxy-1-b daunorubicin are obtained in the form of the chlorohydraph, which is a red. powder is: [a] ^ ° = +255 - 18 ° (c = 0.216, methanol)

Hf = 0.66 [silica gel; methylene chloride - methanol - formic acid - water (88 - 15 - 2 - 1 in vol.)] ..

25 Analysis% calculated: C 56.90 H 6.2b Cl U, 31 H 3 .0% found C 56, ¼ H 6.5 Cl N 3.5 30 Example ΙΓ

In a dry 3-neck round-bottom flask, 1.33 g of L-1 [[biphenylyl-1-propyl-2] oxy-3-carbonyl-leucine are dissolved in 120 cm of anhydrous tetrahydrofuran under an argon atmosphere and then added 0.505 cm 00 1raliriles stored triethylamine

O

7905098; v.: ___ _. · · · · "-: · ·. - 6 -.: · • i

..: - · I

: please. I

t ϊ Λ,? It is cooled to -20 ° C and 0.426 cur is added quickly; isobutyl chloroformate. Shake at -20 ° C for 30 minutes,> 3

240 cm are then added to the white suspension. methylene chloride, 0.42 cm dried triethyl amine over potassium carbonate and 2.14 g; (dithiolan-1,3 yl-2) carhonyloxy-14 daunoruhicin chlorohydrate. in powder form, | It is stirred at -20 ° C for 45 minutes.

_. 3 ..

The reaction mixture is poured from 300 ml of ice-10 water; it is decanted and the organic phase is washed with two times 250! * 3 3; cnr 0.0 5N chlorine water or acid and then with three times 250 cm of water ; The organic solution is dried: over sodium sulphate, 1-yphate, filter and concentrate to dryness under reduced pressure. (20 mm. | 15 ί mercury) without coming to an oven at 30 ° C "- j

The solid residue is dissolved in a mixture of

. * I

methylene chloride that is 25% ethyl acetate. (by volume) and chromatographed on 140 g of silica Merck 0.04-0.063 mm, under Q, 5 nitrogen, The elution is followed by thin layer chromatography. it. Corresponding eluate of the pure product is concentrated to dryness under reduced pressure (20 mm. mercury) and a rent temperature of 300 ° C.

After drying at 20 ° C under reduced pressure (1 mm of mercury), 2.44 g of N— [N - {(hiphenylyl-4) -2-propyl-2] oxycarhonyl} L-leucyl] (dithiolan-1, 3 yl-2] carhonyloxy-14 daunoruhicin in the form of a red powder.

2.44 g of N- [N - {[(diphenylyl-4) -2-propyl-2] -oxycarhonyll L leucyl] (dithiolan-1,3 yl-2) carbonyloxy-14 daunorufai- are dissolved

3Q cine in 150 ml of anhydrous methylene chloride. It is cooled to 5 ° C

3 and then rapidly add 6.88 cm. Of a 0.77 µl dry hydrogen chloride solution in dioxane.

The mixture is stirred for 10 minutes and then the precipitate formed is filtered off. The solid was washed with 100 ml of methylene chloride. The insoluble material is dissolved with 50 cm 7905098 3 anhydrous methanol and subsequently reprecipitated by adding 150 cm. ethyl ether.

After filtration, washing with ethyl ether and drying under low pressure (1 mm of mercury), 1.3 g of N (L-1sucyl) - (di-5 thiolan-1,3-yl-S)} carbonyloxy-1'-daunorone is obtained. c in e-chloro orhydrate.

Rf: 0.57 [silica gel; methylene chloride-methanol-formic acid-water (88-15-2-1 per volume)] *

Analysis.

% her. C 53 * 81! H 5.50 Cl k, 30 E 3.39 S T »7T 10 * Sev * 53.9 5Λ. K9 3.2 7.6

The invention also includes the pharmaceutical preparations consisting of a compound of the general formula 1 in free, or in. the form of the salt, as such or in the form of a medicinal mixture, containing hair, combined with one or more diluents or compatible and pharmaceutically acceptable additives, which may be inert or physiologically active.

These mixtures can be presented in any form suitable for the intended administration. The parenteral route is preferred, especially the intravenous route.

The mixtures of the invention for parenteral administration may or may not be aqueous sterile solutions, suspensions or emulsions.

As the solvent or carrier, mention may be made of propylene glycol, polyethylene glycol, vegetable oils, in particular olive oil and the injectable organic esters, such as ethyl oleate.

These mixtures can also contain additives, in particular wetting, emulsifying and dispersing agents.

Sterilization can be done in various ways, e.g. with the aid of a bacteriological filter, by incorporation into the mixture of sterilizing agents or by irradiation.

They can also be prepared in the form of sterile solid mixtures, which can be dissolved or dispersed in sterile water or any other sterile medium at the time of use.

The compounds of the general formula I and their salts have been used in particular for the treatment of acute lymphoblastic and myeloblastic leukemias, lymph osarcoma, Hodgkin's disease and solid tumors (lung cancer, breast cancer, cancer); 8 -..

ί; '; of the digestive tract and metastases) in daily doses:. generally between 1 and 5 mg / kg intravenously for an adult. ] j The following example illustrates such a mixture.

i 5 Example 3

A solution containing 27.9 mg / cm is prepared; : N— (E. leucyl) diethoxyacet0x7-1 daunorubicin chlorohydrate by pressing—! ! dissolving g of this compound in an apyrogenic physiological solvent in an amount sufficient to obtain

3 ... I

1Q i of 150 cm. The solution obtained is aseptically introduced with ampoules. a quantity of 3 cm / ampoule. The ampoules are sealed and each contain 80 mg 1T- (Leucyi): diethoxy acetoxy-1-dauno-j.

I | ; ruhioine (base) ..

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l 7905098

Claims (4)

1. Renew daunorubic in derivative, from the general formula 1 of the formula sheet, in which the symbol represents a formula which is all of the general formula 2 of the formula sheet, in which the symbols X, which represent oxygen or sulfur atoms, are identical and the symbols R ^ radicals represent alkyl, phenyl or p-position (substituted by a methyl, methoxy or methylthio group) phenyl, or together form an alkylene radical with 2 to 4 carbon atoms, or the symbols T different and the symbols R, together a. J i alkylene radical. with 2 to 4 carbon atoms and the symbol Rg '10 represents a radical of the general formula 3 of the formula sheet, wherein S 1 represents an atom, hydrogen or an alkyl, phenyl, or j benzyl radical, where. the above-mentioned alkyl radicals or parts! straight or branched and containing -1 to 4 carbon atoms are diastereo-; isomeric forms and bun mixtures, as well as the addition salts with acids. ; 2. Method for preparing. a compound according to claim 1, characterized in that a reactive amino acid derivative of the general formula 4, wherein. is as defined above and the amine function of which has been previously protected, reacting with a daunorubive derivative of the general formula 5, wherein 2Q Rj is as described above, after which the protective group is removed and the resulting compound optionally converted into an acid addition salt . 3. Process according to claim 2, characterized in that as reactive amino acid derivative of the general formula 4, wherein R 2 is as described above, a mixed anhydride or a reactive ester, the amine function of which is protected, is reacted with a daunorubicin derivative of the general formula 5, wherein R 2 is as described above, after which the protecting group is removed and the resulting compound optionally converted into an acid addition salt. 4. Process according to claim 2, characterized in that a reactive amino acid derivative of the general formula 4 of the formula sheet, wherein RT is as defined above and of which the amine-4? V905098 V. * ·., - ...... ft - -. -10- / K: function is pre-protected, by a ^ biphenylyl - ^) - 2-propy 1-2] -. oxycarbonyl or -trytyl react with a daunorubic in derivative of. the general formula, in which. is as described above, after which the protecting group is removed and the resulting compound is optionally converted into an acid addition salt. 5 * Medicament »consisting of a compound according to. Claim 1 in pure form or, in the form of. a hair-containing pharmaceutical mixture in combination with one or more tolerant and pharmaceutically acceptable diluents or additives. i | i i | ? »*’ - i \. | T \ © V 7905098. .,. ... 1 OH 1 -. II CO CH, OCOR, -. ^ / Yy ^ Uh Λ W * *: \ vw r * CH30 0 .OH 0 - CH - CH2 - CH - CHOH - CH - CH3 I— _i 2 x-r3 - CH <Y ^ x-r3 3 R-GH - CO - 4 I nh2 - V "------------------------- - - = 1 R, _CH_COOH. 4 I NH2 0 OH CO CH2 OCOR1 5 -0H YyVy r =: -..... CH, 0 0 OH 0 - OH. — CH, - CB - OHOH - CH - CH, I-2-o _-_ l 3 7905098