FR2774989A1 - New dolastatin conjugates useful as anticancer agents, especially against carcinomas with high levels of steroid hormone receptor expression, e.g. ovarian and breast cancer - Google Patents

Abstract

Estradiol-dolastatin conjugates (I) are new. Estradiol-dolastatin conjugates of formula (I) are new: A = an estradiol group of formula (i): R1 = H, 1-6C alkyl, 2-6C alkenyl, 1-6C acyl, arylcarbonyl or aryl(1-6C)alkylcarbonyl; aryl = phenyl or naphthyl optionally substituted by halogen, 1-6C alkyl, 1-6C alkoxy, 1-6C acyl, CN, NO2, NH2, 1-6C alkylamino or di(1-6C alkyl)amino; R2 = H, 1-6C alkyl, 1-6C acyl, arylcarbonyl, aryl(1-6C)alkylcarbonyl or a bond to D; R3 = H when R2 is a bond to D, or a bond to D when R2 is H, 1-6C alkyl, 1-6C acyl, arylcarbonyl or aryl(1-6C)alkylcarbonyl; D = -X-Y-Z-, with X bonded to E and Z bonded to A; X = 1-6C alkylene, -CO-Ra- or -P(O)(Rb)-O-Ra-; Ra = 1-6C alkylene; Rb = 1-6C alkyl; Y = CO or NHCO with the carbonyl group bonded to Z; Z = a single bond, -NH-Ra- or -Ra-CO-; E = a dolastatin-15 group of formula (ii) or a dolastatin-10 group of formula (iii): R4 = isopropyl or CH(Me)O with the O atom bonded to X: R5 = a group of formula E1 or E2 when R4 is isopropyl or a group of formula E1' or E2' when R4 is CH(Me)O: R6 = H, 1-6C alkoxy or an O atom bonded to X; R7 = an O atom bonded to X when R6 is H or alkoxy, or 1-6C alkoxy when R6 is an O atom bonded to X: R8 = an O atom bonded to X; n = 1-4; R'6 = H or 1-6C alkoxy; R'7 = 1-6C alkoxy; R'8 = H or 1-6C alkoxy; R9 = a group of formula E3 or E4 when R4 is isopropyl, or a group of formula E3' or E4' when R4 is CH(Me)O: R10 = -CO-O- with the O atom bonded to X, or -CO-NH- with the N atom bonded to X; R11 = an O atom bonded to X; R'10 = H; R'11 = H or 1-6C alkoxy. An Independent claim is included for methods of preparing (I).

Description

The present invention relates to novel cytotoxic compounds derived

estradiol.

  The targeting of cancer cells by cytotoxic molecules aims to reduce the side effects caused by agents used in cancer therapy. Various strategies have

  have been studied and implemented for the targeting of drugs against tumor cells.

  In the context of the invention described by the Applicant, the cytotoxic compound is bound in it being of steroid type, the corresponding receptor is expressed in rapidly developing cells. The compounds of the invention are thus steroid-cytotoxic hybrid molecules, the latter part consisting of derivatives of various

  dolastatins. These dolastatin are pseudopeptides with strong antiproliferative activity.

  The compounds of the invention, in addition to being novel, surprisingly exhibit a significant and encouraging in-vitro activity. in terms of their specificity of action. The new compounds, presented by the Applicant, thus have antitumor properties which make them particularly suitable for use in the treatment of cancers and in particular carcinomas characterized by high levels of expression of steroid-type hormone receptors, such as tumors. ovarian or

breast tumors.

  The present invention relates to the compounds of formula (I):

(E) - (D) - (A) (I)

  in which: (A) represents an oestradiol derivative, of formula: OR Me 2 R 3 R 10 in which R 1 represents a hydrogen atom, a linear or branched (C 1 -C 4) alkyl group, an alkenyl group (C2- C6) linear or branched, a linear or branched (C1-C6) acyl group, an arylcarbonyl group or an arylalkylcarbonyl group in which the aryl part represents a phenyl group or a naphthyl group (these groups being optionally substituted by at least one atom of halogen, a linear or branched (C1-C6) alkyl group, linear or branched (C1-C6) alkoxy, linear or branched (C1-C6) acyl, cyano, nitro, or an amino group, itself optionally substituted by one or two linear or branched alkyl (C1-C6) groups) and the linear or branched alkyl part comprises

from 1 to 6 carbon atoms.

  R2 represents a hydrogen atom, a linear or branched (C1-C6) alkyl group.

  a linear or branched acyl (Cl-C6) group, an arylcarbonyl group or an arylalkylcarbonyl group having the same definition as in R1, or consists of a single bond (said bond corresponding to that existing between the part (A) and the part ( D) compounds of formula (I)), R3 represents a hydrogen atom when R2 represents a single bond or R3 represents a single bond (said bond corresponding to that existing between part (A) and part (D) of compounds of formula (I)) when R2 has one of the other

  definitions previously described.

  (D) represents a linear or branched, polysubstituted chain of formula:

-X Y Z-

  in which X is connected at its free end to the part (E) of formula (I) and Z is connected at its free end to part (A) of formula (I) and where: X represents: - a group linear or branched (C 1 -C 6) alkylene, O il - a carbonylalkylene group -CR- in which Ra represents a linear or branched (C 1 -C 6) alkylene group, O - or a group - POR - in which Ra represents a grouping R a is linear or branched (C1-C6) alkylene and Rb represents a linear or branched (C1-C6) alkyl group, Y represents a carbonyl group or an amido-NH-C- group in which the carbonyl group is bonded; in part Z of group (D) of formula (I), Z represents: - a single bond - an aminoalkylene -NH-Ra- group in which Ra represents a linear or branched (C1-C6) alkylene group, - or an alkylene carbonyl group -R / -C- in which Ra has the same garlic

O

  definition as before, (E) represents: * a derivative of Dolastatin-15 of formula: R4 Me

NN N

  In which: R4 represents: * is an isopropyl group and in this case: R5 represents: - a group of formula (E1): R6 -O N R7 (E1)

O

  in which: R6 represents a hydrogen atom, or a linear or branched (C1-C6) alkoxy group, and in this case R7 represents an oxygen atom, said oxygen atom being connected to the group X of the part ( D) of the formula (I): 5. or R6 represents an oxygen atom, said oxygen atom being connected to the group X of the part (D) of the formula (I), and in this case R7 represents a linear or branched (C1-C6) alkoxy group, or a group of formula (E2):

-NH- (CH2) R-- (ER)

R (E2)

  in which: R 8 represents an oxygen atom, said oxygen atom being connected to the X group of the part (D) of the formula (I), n represents an integer between 1 and 4 inclusive, * is a group of CH-CH-O- formula in which the oxygen atom is

1

  connected to the group X of part (D) of formula (I), and in this case: Rs represents: a group of formula (E '): R'6 -ON R7 (E') OO in which: R6 represents a hydrogen atom, or a linear or branched (C1-C6) alkoxy group, R'7 represents a linear or branched (C1-C6) alkoxy group, or a group of formula (E2 '): or in wherein R6 'represents a hydrogen atom, or a linear or branched alkoxy (C1-C4) group, R'7 represents a linear or branched alkoxy (C1-C4) group, or a group of formula (ED' ) -NH- (CH 2) n -R'8 (E2) in which: R'8 represents a hydrogen atom, or a linear or branched (C1-C6) alkoxy group, n represents an integer between 1 and 4 inclusive, * or a derivative of Dolastafine-10 of the formula: R4 H o Me HsI

N R

N - N9

  In which: R4 represents: * is an isopropyl group and in this case: R9 represents: a group of formula (E3): ## EQU1 ##

-N X), (E3)

\ Born

  In which: RWo represents a group -C-O- (in which the oxygen atom is connected

O

  to the group X of part (D) of formula (I)) or a group -CNH-

  it (in which the nitrogen atom is connected to the group X of the part (D) of the formula (I)), or a group of formula (E4): -NH- (CH,) - Ril ( E4) in which: Rn represents an oxygen atom. said oxygen atom being connected to the group X of the part (D) of the formula (I).

  n represents an integer between 1 and 4 inclusive.

  or a group of formula CH - HO - in which the oxygen atom is connected to the group X of part (D) of formula (I), and in this case: R 9 represents: a group of formula (E'3):

-NS S_ (E'3)

  Wherein R'10o represents a hydrogen atom, - or a group of formula (E'4): - NH- (CH2) n --- R'11 (E'a) in which R Represents a hydrogen atom or a linear or branched (C1-C6) alkoxy group n represents an integer between 1 and 4 inclusive, their isomers, as well as their addition salts with an acid or a pharmaceutically acceptable base

acceptable.

  Among the pharmaceutically acceptable acids, mention may be made, without limitation, of hydrochloric, hydrobromic, sulfuric and phosphonic acids. acetic, trifluoroacetic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, tartaric, maleic, citric,

  ascorbic, oxalic, methanesulfonic, camphoric, etc ...

  Among the pharmaceutically acceptable bases, mention may be made, without limitation

  sodium hydroxide, potassium hydroxide, triethylamine, tertbutylamine, etc.

  The preferred substituent R4 according to the invention is the isopropyl substituent. Advantageously, the preferred compounds of the invention are those of formula (I) in which: - (E) represents a derivative of Dolastatin-15 in which the group R5 represents a group of formula (E1) such that defined above, or (E) represents a derivative of Dolastatin-10 in which the R9 group represents a group of formula (E3) as defined above, their isomers and their addition salts with an acid or with a base pharmaceutically

acceptable.

  According to a particularly advantageous variant of the invention, the preferred compounds of the invention are those of formula (IA), in particular the compounds of formula (I):

/ 7 / O-X-Y-Z-O

  R4 me SC H0 0f R7 R | Embedded image in which R 1 is as defined in formula (I) and preferably represents a hydrogen atom, a linear or branched (C 2 -C 6) alkenyl group, or an arylcarbonyl group. said group being advantageously a benzoyl group, Rs represents a hydrogen atom, R4 represents an isopropyl group, R7 represents a linear or branched (C1-C6) alkoxy group, and preferably a methoxy group, OX represents a carbonylalkylene group; -C- R-, the carbonyl part being at O connected to the neighboring oxygen atom, or a group of formula -POR-, to S Rb the phosphorus atom being connected to the neighboring oxygen atom, groups in which Ra represents a linear or branched (C1-C6) alkylene group and preferably of linear structure, and Rb represents a linear or branched (C1-C6) alkyl group, and particularly preferably, a methyl group, O Y represents a carbonyl group -C-, and Z represents a single bond, their isomers and their addition salts with an acid or a pharmaceutically acceptable base;

acceptable.

  According to another advantageous variant of the invention, the compounds of the invention are those of formula (IB), in particular the compounds of formula (I):

R4 H R6

H O 0OX ---

  Embedded image in which: R 1 is as defined in formula (I), wherein R 1 is as defined in formula (I) ) and preferably represents a hydrogen atom, a linear or branched (C 2 -C 6) alkenyl group, or an arylcarbonyl group, said group being advantageously a benzoyl group, R 3 represents a hydrogen atom, R 4 represents an isopropyl group , R6 represents a hydrogen atom, X represents a linear or branched (C1-C6) alkylene group and preferably a linear (C1-C6) alkylene group, Y represents an amido -NH-C- group in which the part -NH is connected to the part (X), Z represents an alkylene carbonyl group -RC- in which the O carbonyl part is connected to the neighboring oxygen atom, and Ra represents an alkylene group (C1- C6) linear or branched, and preferably Ra is linear, their isomers and their addition salts to an acid e or a pharmaceutically based

acceptable.

  According to a third advantageous variant of the invention, the preferred compounds are those of formula (IC), in particular the compounds of formula (I): ## STR2 ##

/ \ O (IC)

  in which: R 1 is as defined in formula (1) and preferably represents a hydrogen atom, a linear or branched (C 2 -C 6) alkenyl group, or an arylcarbonyl group, said group being advantageously a benzoyl group, R 2 is as defined in formula (I) and preferably represents a hydrogen atom or an arylcarbonyl group, said group advantageously being a benzoyl group, R 4 represents an isopropyl group, R 7 represents an alkoxy group (C 1 -C 6) linear or branched and preferably a methoxy group, OX represents a carbonylalkylene -CR- group. the carbonyl part being C-Ra O

  connected to the neighboring oxygen atom, or a group of formula -P-O-R-

  a Rb the phosphorus atom being connected to the neighboring oxygen atom, groups in which Ra represents a linear or branched (C1-C6) alkylene group and preferably Ra is linear, and Rb represents an alkyl group (C1 -C6) linear or branched, and particularly preferably Rb represents a methyl group, Y represents a carbonyl group, Z represents an aminoalkylene -NH-Ra- group, the amine part being connected to Y and Ra representing an alkylene group (CI -C6) linear or branched, their isomers and their addition salts with an acid or a pharmaceutically acceptable base

acceptable.

  The present invention also extends to the process for preparing the compounds of formula (I), characterized in that the starting material used is an oestradiol derivative of formula (II): ## STR1 ## (II) R10 wherein R1 and R2 have the same definition as in formula (I) with the proviso that R2 is different from a single bond. and R'3 represents a hydrogen atom or a linear or branched (C1-C6) aminoalkyl group, compounds of formula (II) which are reacted, in the ca.so R'3 and R2 represent simultaneously a hydrogen atom, under basic conditions, in the presence of a coupling agent, with a compound of formula (III):

0 0

  Wherein R a is as defined in formula (I) and R c is a protecting group of carboxylic acid functions, to give compounds of formula (IIa),

O O

  (Ila) RiO in which RI, Ra and R1 have the same definition as above, compounds of formula (IIa) whose terminal carboxylic acid function is deprotected according to standard deprotection conditions of these functions to yield compounds of formula (IIb) :

O 0

  Wherein R1 and Ra have the same definition as above, compounds of formula (IIb) which are coupled: either with a derivative of Dolastatin-15 of formula (IV / a) ## STR3 ## wherein R 5 represents a group of formula (E ') or (E), wherein R 5 represents a group of formula (E') or (E 2) as defined in the general definition of formula (I), to give compounds of formula (I / a), a particular case of compounds of formula (I):

O O

  Embedded image in which R 1 and R a are as defined above and R 5 is as defined in formula (IV / a); or with a derivative of Dolastatin-15 of formula (IV / b): ## STR2 ## (IV / b)

N N N '

/ II: I, III - '

  Wherein R7 represents a linear or branched (C1-C6) alkoxy group to give compounds of formula (I / b), a particular case of compounds of formula (I) ## STR2 ## Wherein R1 and Ra are as defined above and R7 is as defined in formula (IV / b) above: either with a derivative of Dolastatin-15 of formula (IV / c): Me NNXfJk N NH- (CH2) OH

N - N

  In which n is an integer between 1 and 4 inclusive, to give compounds of formula (I / c), a particular case of compounds of formula (I):

/ A0 00

  Wherein n, R 1 and Ra are as previously defined, with either a derivative of the formula ## STR1 ## wherein Dolastatin-10 of formula (V / a):

142774989

  Embedded image in which R 9 represents a group of formula (E '3) or (E' 4) as defined in the general definition of the formula ## STR3 ## wherein R 9 represents a group of formula (E '3) or (E' 4) as defined in the general definition of the formula (I), to give compounds of formula (I / d), a particular case of compounds of formula (I):

O O

Oe ito H O M Me Me

/ N R

N-N 9

  Embedded image in which R 1 and R a are as defined above and R 9 is as defined in formula (V / a), or with a derivative of Dolastatin-10 of formula (V) / b): Me / H OMe Me

/ N X N XNIX A <NH- (CH,) OH

Me O / \ Me Me OMe OMe (V / b)

  in which n is an integer between 1 and 4 inclusive.

  to give compounds of formula (I / e), a particular case of the compounds of formula (I): Me, Ho MN, HXpUN NH- (CH) N Me O Me O Me O Me (I / e) ORI in which n. R1 and Ra are as defined previously, * or with a compound of formula (VI): R6 Me 'N0 I "/ R-NH

N N

N N N

  O / Me OO (VI) in which Ra is as defined in formula (I) and R6 represents a hydrogen atom or a linear or branched (C1-C6) alkoxy group, said compounds of formula (VI) being obtained by reaction. in the presence of triphenylphosphine and diethylazodicarboxylate, between a compound of formula (VI / a): ## STR2 ##

a 2--

  wherein Ra is as defined above, and a compound of formula (IV / d), derived from Dolastatin-R6 MeY H O OH I II II N ,, k, N 0

N O N>

 N N

  Wherein R6 represents a hydrogen atom or a linear or branched (C1-C6) alkoxy group, to yield compounds of formula (VIIb): R6 o

  ## STR2 ##

  ## STR5 ## wherein Ra and R6 are such that defined above, compounds of formula (VI / b) whose carbamrnate function is reduced by hydrogenolysis in the presence of Pd / C, to give the compounds of formula (VI) defined above, said compounds of formula (VI), put into the presence of said compounds of formula (IIb) previously described, in basic medium and in the presence of a coupling agent such as tris (dimethylamino) bromophosphonium hexafluorophosphate, lead to compounds of formula (If), a particular case of compounds of formula (I): No.

O / \ MeO o o.

/ \ O NHO (I / f) - Me OR1

  wherein R1, R6 and Ra are as previously defined.

  or compounds of formula (II) which are reacted, in the case where R 'and R2 simultaneously represent a hydrogen atom, X is with a compound of formula (VII): ## STR2 ## Rb in which Ra and Rb are as defined in formula (I) and W represents a derivative of Dolastatin-15 or Dolastatin-10, as defined below, said compounds of formula (VII) being obtained by addition in basic condition of a compound of formula (VI / a): O o

0 1

  HO-PO-Ra OR (VII / a) Rb in which Ra and Rb are as defined above and RK represents a protecting group of the carboxylic acid functions, such as a benzyl group, with one of the compounds of formula (IV /at). (IV / b), (IV / c), (V / a) and (V / b) at their free alcohol function, to yield compounds of formula (VIIb). in which Ra, Rb and RK are as defined above and W represents one of the residues of the compounds of formula (IV / a), (IV / b), (IV / c) ), (V / a) or (V / b) in which the hydrogen atom of the free alcohol function is substituted by the phosphorus atom of said compounds (VIIb), compounds of formula (VIIb) whose acid function terminal carboxylic acid is deprotected to yield compounds of formula (VII), said compounds of formula (VII). brought into contact with said compounds of formula (I) under basic conditions and in the presence of isopropenyl chlorocarbonate, give compounds of formula (I / g), a particular case of compounds of formula (I):

W-P-O-R O

  where RI, Ra and Rh are as defined above and W represents one of the residues of formula (IV / a), (IV / b), (IV / c). (V / a) and (V / b) in which the hydrogen atom of the alcohol function is substituted by the phosphorus atom of the compounds (I / g), X or with a derivative of Dolastatin-10 of the formula (Vc): Me HO \ H) Me I

K NH

N Pl Me I IMe OMe O OMe S

O OH

  wherein Ra is as defined above and M represents an oxygen atom or a group -NH-. said compound of formula (Vc) being obtained by addition reaction, under basic conditions and in the presence of a coupling reagent, of a compound of formula (VIII): ## STR2 ## which Ra and Rc are as defined above and M 'represents a hydroxyl group or an amino group, with a derivative of dolastatin 10 of formula (Vd): ## STR1 ## O0 to give the compounds of formula (Ve): ## STR2 ## in which Ra, R and M are as defined above, compounds of formula (Ve), the terminal carboxylic acid function of which is deprotected under conventional conditions to yield the compounds of formula (Vc) previously described, said compounds of formula (Vc), brought into contact with said compounds of formula (II) under condition in the presence of isopropenyl chlorocarbonate, for example, lead to compounds of formula (1 / h), a particular case of compounds of formula (I): ## STR1 ## Me O - Me OMe O OMe O

aminoalkyl group NH 2 -Ra

M1 M-R11 (1) h)

0- M-R "'

  ORI dan is with a derivative of formula nt such as challengeinii previously, in the presence of a base such as triethylamine and a coupling agent, to lead to the compounds of formula (that one makes ragir, in the case om R'3 represents a ## STR2 ## in which Riv R2, Ra and R are (V as defined above and W represents one of the residues of formula (Vta), (IV / b), (V / c), (V / a) e and (V / b) including the oxygen atom of oupling,

  For the compol is bound to the phosphorule atoms of the compounds of formula (I).

O O OR2

W-P - O-R - H

  Wherein R 1, R 2, R a and R b are as previously defined and W represents one of the residues of formula (IV / a), (IV / b), (IV / c), (V / a) and (V / b) in which the oxygen atom of the alcohol function is bonded to the phosphorus atom of the compounds (I / i),

2774989

  * with a derivative of formula (IX).

O

W-C-R. OH (IX)

  in which Ra and W are as defined above and said compounds of formula (IX) being obtained by addition reaction under basic conditions, and in the presence of a coupling agent, of a compound of formula (E) such that defined above with one of the compounds of formula (IV / a). (IV / b). (IV / c), (V / a) and (V / b) in their free alcohol function. to give the compounds of formula (IXa): wherein W represents one of the residues of the compounds of formula (IV / a), (IV / b). (IV / c),

  (V / a) and (V / b) and Ra and R1 are as previously defined.

  compounds of formula (IXa), the carboxylic acid function of which is deprotected under standard deprotection conditions in order to obtain the compounds of formula (IXa) as described, said compounds of formula (IX), brought into contact with said compounds of formula (II) , in

  basic condition in the presence of triethylamine for example, and a coupling reagent.

  lead to compounds of formula (I / j), a particular case of compounds of formula (1)

O O OR2

  in which R 1, R 2, Ra and W are as defined above, or with a derivative of formula (Vc) as defined above, to give compounds of formula (I) I / k), a particular case of the compounds of formula (I): Me-, Me

M - H N N

e Me M () Me o OMe o

> NH-R

  (I / k) wherein R1, R2, Ra and M are as defined above, the compounds (I / a) to (I / k) forming all of the compounds of the invention, which are purified, if appropriate, according to a conventional purification technique, which can, if desired, be separated into their different isomers according to a conventional separation technique, and which, if appropriate, are converted into their salts; addition to an acid or a base

pharmaceutically acceptable.

  The compounds of formulas (I), (III), (Va) and (VIII) are either compounds

  are obtained according to conventional methods of organic synthesis.

  Compounds of formula (IV / a), (IV / b), (IV / c), (IV / d), (V / a), (V / b) and (V / d), derived from dolastatin -15 or dolastatin-10, are obtained according to conventional methods of amino acid coupling, by successive reaction between an amino acid whose terminal carboxylic acid function is protected and the primary amine function is free and another amino acid whose amine function terminal is protected and the carboxylic acid function is free. The C-terminal aromatic part is also introduced according to this same coupling strategy. This synthesis strategy is notably described in the following articles: Tetrahedron 1992, 48, 4115-4122 and J. Am. Chem. Soc. 1991, 113, 6692-6693, as well as in

patent application WO 93/23424.

  The compounds of formula (I) have particularly advantageous antitumor properties. They have excellent cytotoxicity in vitro on cell lines, and an action on the cell cycle. In addition, these compounds of formula (I) have greater activity specificity than Dolastatin-15 on cell lines whose growth is estrogen-regulated. The characteristic properties of these compounds allow their

  therapeutic use as antitumor agents.

  The subject of the present invention is also the pharmaceutical compositions containing the products of formula (I), their optical isomers or one of their addition salts with a base or a pharmaceutically acceptable acid, alone or in combination with one or more

  Inert or non-toxic excipients or vehicles.

  Among the pharmaceutical compositions according to the invention, mention will be made more particularly of those which are suitable for oral, parenteral (intravenous, intramuscular or subcutaneous), per-transcutaneous, nasal, rectal administration. perlingual, ocular or respiratory. and especially single or coated tablets, sublingual tablets. sachets, capsules, glossettes, lozenges, suppositories, creams, ointments, dermal gels, injectable or drinkable preparations, aerosols, eye drops or

nasal, etc.

  The dosage varies with the age and weight of the patient, the route of administration, the nature and severity of the condition, and any associated treatments and ranges from

  0.1 mg / m2 to 50 mg / m2 in one or more doses per day.

  The following examples illustrate the invention but do not limit it in any way.

  The starting materials used are known products or prepared according to

known operating procedures.

  The structures of the compounds described in the examples and the preparations were determined according to the usual spectrophotometric techniques (infrared, magnetic resonance,

  nuclear, mass spectrometry, ...).

  Example 1 (5S) -I - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-

  prolyl-L-prolyl] -2-hydroxyisovaleryl] -2-oxo-4-methoxy-5 - [[4- (17 [(malonyloxy) -3-benzoyl-estradiol) phenyl] methyl] -3-pyrroline = Dol -PhO-MAy-OES17Bz

0 0

    Step A: Synthesis of Pentapeptide Bn-OC-Val-Val-MeVal-Pro-Pro-OH Step 1: General Method of Coupling to a Cooled Solution at A 0 C of the D-Salt ammonium of the amino acid (1 equivalent) in dichloromethane. are successively added the amino-acid-N-protected by a group (C (O) OBn) (1.2 equivalents), 2.5 equivalents of N.N-

diisopropyl-N-

  ethylamine and 1.2 equivalents of the coupling agent, namely PyBroP (tripyrrolidinobromophosphonium hexafluorophosphate). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure. The residue obtained is taken up with ethyl acetate. then washed with aqueous solutions of KHSO4, NaHCO3 and saturated with NaCl to neutral pH. The organic phase is then dried over Na 2 SO 4, filtered and concentrated under pressure.

  scaled down. The crude product obtained is chromatographed on silica gel.

  Step 2: general method of hydrogenolysis The N-protected amino acid is solubilized in a mixture of methanol and 3N HCl. This solution is hydrogenolysed on Pd / C (20% by weight). The solution is then filtered

  and concentrated under reduced pressure.

  By successive combination of stage I and stage 2, are thus coupled 5, 92 g of L-

  Proline-OtBu (whose terminal carboxylic acid function is protected by a t-butyl group) with 4.12 g of Bn-O-Proline whose N-terminal amine function is protected by a benzyloxycarbonyl group, this dipeptide then being hydrogenolysed to be coupled with 4.7 g of LN-Methyl-Valine whose amine function is protected by a benzyloxycarbonyl group. then after hydrogenolysis, the tripeptide is coupled with 3.4 g of L-Valine whose terminal amine function is protected by a benzyloxycarbonyl group. then after hydrogenolysis, the tetrapeptide is coupled with 0.9 g of L-Valine whose terminal amine function is protected by a benzyloxycarbonyl group. 1.7 g of pentapeptide of formula Bn-O-C-Val-Val-MeVal-Pro-Pro-OtBu are thus obtained. Step 3: Deprotection of the C-terminal Part The previous pentapeptide is dissolved in 3 ml of dichloromethane to which 3 ml of trifluoroacetic acid is added. After 2 hours, the solution is concentrated under pressure

  reduced and the residue is triturated in ether to isolate the expected product.

Melting point: 95-102 C.

  Stage B '(5S) -1 - [(2S) -2-hydroxyisovaleryl] -2-oxo-4-methoxy-5 - [(4-benzyloxy) -4-

  phenyl) methyl-3-pyrroline HO "OBn N / N OMe

O /

  Step 1: Methyl N - [(2S) -2-hydroxy-isovaleryl- (O-benzyl) -L-tyrosinate To a solution of 14.5 g of methyl (O-benzyl) -L-tyrosinate in 100 ml of

  dichloromethane are added 3.7 g of hydroxyisovaleric acid, 18.8 ml of N, N-

  diisopropyl-N-ethylamine and 16.3 g of PyBOP (1H-hexafluorophosphate

  benzotriazol-1-yl-1-oxy) -tripyrrolidinophosphonium). After 18 hours. the solution is concentrated, diluted with ethyl acetate, washed to neutral pH, dried over sodium sulphate and then concentrated under reduced pressure. Chromatography on silica gel

allows to isolate the expected product.

Melting point: 135-137 ° C.

  Step 2: N - [(2S) -2-tert-Butyldimethylsilyl-2-oxy-isovaleryl] - (O-benzyl) L-tyrosine A 10.1 g of the product of step 1 in 30 ml of DMF are added 5.9 g of chloride of

  t-butyldimethylsilyl and 4.5 g of imidazole. After 12 hours, the solution is concentrated.

  s 2774989 diluted in ethyl acetate. washed with an aqueous KHSO4 solution, dried over Na2SO4 then filtered and concentrated under reduced pressure. The residue is then diluted with 1 ml of dioxane and 52 ml of 1N sodium hydroxide are added. After 4 hours, the reaction is neutralized with KHSO4 and then diluted with ethyl acetate. After washing, drying and filtering the organic phase, it is concentrated under reduced pressure. The residue

  crystallizes in ethyl ether to isolate the expected product.

Melting point: 127-129 C.

  Step 3: (5S) -I - [(2S) -2-tert-Butyldimethylsilyl-2-oxy-isovaleryl-2-oxo-4-hydroxy-5-

  [(4-benzyloxyphenyl) methyl] -3-pyrroline To a solution of 5.6 g of the product of step 2 in 25 ml of dichloromethane, 3.5 g of dimethylaminopyridine 3.3 is added at -10 ° C. g of Meldrum acid and then dropwise 1.5 ml of isopropenyl chlorocarbonate diluted in 2 ml of dichloromethane. After 4 hours of reaction, the organic phase is washed, dried over Na 2 SO 4, filtered and concentrated under reduced pressure to obtain an oil which is dissolved in 20 ml of acetonitrile and refluxed for 20 minutes. After

  evaporation of the solvent, the expected product is obtained in the form of a gum.

  Step 4: (5S) -1- [(2S) -O-tert-Butyldimethylsilyl-2-hydroxy-isovaleryl-2-oxo-4-

  Methoxy-5-I (4-benzyloxy-phenyl) methyl-3-pyrroline A 2.3 g of the compound of step 3 dissolved in 10 ml of THF are added 1.4 g of PPh 3, 0.22 ml of methanol and then 0 85 ml of diethyl azodicarboxylate (DEAD). After minutes, the solvent is evaporated. The residue is diluted in ethyl acetate and the organic phase is washed, dried, filtered and concentrated under reduced pressure. A

  Chromatography on silica gel makes it possible to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z = 524.

  Step 5: (5S) - I - [(2S) -2-hydroxy-isovaleryl-2-oxo-4-methoxy-5 '[(4-benzyloxy)] -

  phenyl) methyl-3-pyrroline To 1.3 g of the compound obtained in step 4 are added 5 ml of trifluoroacetic acid. After 30 minutes of reaction, the reaction is concentrated under reduced pressure and chromatography on silica gel (eluent: ethyl acetate / cyclohexane

  35/65) makes it possible to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z = 410.

  Stage C: (5S) -1 - [(2S) -O - [[(N-benzyloxycarbonyl) -L-valyl] -L-valyl- (N-methyl-L-valyl) -L

  prolyl-L-prolyl] -2-hydroxy-isoval.érvIl] -2-oxo-4-methoxy-5 - [(4-benzvloxv-

  phenyl) methyl] -3-pyrroline To a solution of 0.5 g of the compound of step 3 of step A and 0.31 g of the compound of step 5 of step B are added to OC 0.2 ml of triethylamine 3.7 mg of DMAP diluted in 10 ml of dichloromethane and 0.16 ml of isopropenyl chlorocarbonate diluted in 1 ml of dichloromethane. After 24 hours at room temperature, the solution is

  diluted with ethyl acetate, washed, dried over sodium sulfate, filtered and concentrated.

  Chromatography on silica gel (ethyl acetate / cyclohexane: 30/70) allows

to isolate the expected product.

Melting point: 107-111 C.

  Stage D: (5S) -1 - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-prolyl-L-

  prolyl-2-hydroxy-isovaleryl-2-oxo-4-methoxy-5 - [(4-hydroxyphenyl) methyl] -3-pyrroline: (Dol-Ph-OH) OH N / N-NX Ok NN Me O 0.4 g of the compound of Stage C dissolved in 10 ml of methanol is hydrogenolyzed on Pd / C in the presence of 1.47 ml of HCHO in aqueous solution. After 18 hours, the reaction is filtered and concentrated under reduced pressure. Chromatography on silica gel (dichloromethane / methanol / Et 3 N 95/5 / 1) followed by lyophilization makes it possible to isolate the

expected product.

  Mass Spectrum: FAB +: [M + H] m / z = 853.

  Stage E: 3-benzoyl-173-malonyl estradiol

277-74989

0 0

MO / LOH

BzO

  To 0.2 g of 3-benzoyl-17β-estradiol at -5 ° C. is added 0.1 g of monobenzylmalonate.

  0.15 ml of triethylamine, 2.6 mg of DMAP dissolved in 5 ml of dichloromethane and 0.11 ml of isopropenyl chlorocarbonate. After 1 hour of reaction with return to ambient temperature, the reaction is diluted with ethyl acetate. The organic phase is washed until neutral, dried over Na 2 SO 4, filtered and concentrated under reduced pressure. The compound thus obtained is then dissolved in a MeOH / THF mixture and 4 equivalents of ammonium formate and 56 mg of Pd / C are added. After 30 minutes of reflux, the reaction mixture is filtered and concentrated under reduced pressure. After dilution of the residue in dichloromethane, washing, drying and concentration, chromatography on silica gel (cyclohexane / ethyl acetate: 60/40) makes it possible to isolate the

expected product.

Melting point: 152-159 C.

  Stage F: Dol-Ph-O-Mal-OES17Bz To 5.3 mg of the compound obtained in stage E dissolved in 1 ml of dichloromethane is added 1 equivalent of the compound obtained in stage D, 2 equivalents of triethylamine and 1 equivalent the PyBOP coupling agent. After 20 hours of reaction at ambient temperature, the solution is diluted with ethyl acetate and washed to neutral pH, then dried, filtered and concentrated under reduced pressure. Chromatography on silica gel (95/5 dichloromethane / methanol) followed by lyophilization makes it possible to isolate

the expected product.

  Mass spectrum: FAB +: [M + H]: m / z: 1297.

  High resolution: ([M + H +]): Found: 1310, 7179

Calculated: 1310, 7328.

  Example 2: (5S) -I - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-

  prolyl-L-prolyl] -2-hydroxyisovaleryl] -2-oxo-4-methoxy-5 - [[4-175B (malonyloxy) -3-allyl-estradiol) phenyl] methyl] -3-pyrroline = Dol-Ph -OMal-OES17-AII oo () O. ## STR2 ## Step A: 3-allyl-1 75-malonyl estradiol

  The procedure is as in Step E of Example 1, using as substrate the 3-

allyl-17 [-estradiol

Melting point: 134-135C.

  Step B: Dol-Ph-O-Mal-OES] 7-All The procedure is as in Step F of Example 1 using as substrate the compound obtained in Step A of Example 2 and the compound obtained in stage D

of Example 1.

  Mass Spectrum: FAB +: [M + H]: m / z: 1341.

  Example 3: (5S) -I - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-

  prolyl-L-prolyl] -2-hydroxyisovaleryl] -2-oxo-4-methoxy-5 - [[4-17, B (malonyloxy) -oestradiol) phenyl] methyl] -3-pyrroline = Dol-Ph- ## STR5 # tetrahydrofuran are added 1.2 mg

  of Pd (PPh3) 4, 11 equivalents of tri-n-butyltin hydride and 11 equivalents of

  nitrophenol. After disappearance of the initial product, the solution is concentrated under reduced pressure and chromatography on silica gel (dichloromethane / methanol) followed by lyophilization makes it possible to obtain the expected product.

  Mass Spectrum: FAB +: [M + H] m / z: 1193.

  Example 4: Dol-Ph-O-Adi-OES17-OBz o Me O o OBz N k0 0, J HN (sJu XOO | M Me, OMe Me O <Me O o, AA Stage A 3-benzoyl-17] - Adipyl estradiol The procedure is as in Step E of Example 1 using as monobenzyl succinate reagent.

  Mass Spectrum: FAB +: [M + H]: m / z: 505.

  Stage B: Dol-Ph-O-Adi-OES / 7-OBz The procedure is as in Stage F of Example 1, using as substrate the compound obtained in Stage A of Example 4 and the compound obtained in stage D

of Example 1.

  Mass Spectrum: FAB +: [M + H]: m / z: 1339.

  Example 5: (5S) -1 - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-prolyl]

  L-prolyl] -2-hydroxy-isovaleryl] -2-oxo-4-methoxy-5 - [[4-173 (3-benzoyl-

  oestradiol) carbonylpropyloxy (methylphosphoryl) -oxy) -phenyl] methyl] -

  3-pyrroline = Dol-Ph-O-Phos-But-OES17-OBz 0o It Me 0 '| 0

0 & t Me -

* OBz Me t OMe

N N 0 N

Me O / AMe o O / \ A

  Step A: (55) -] - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valvl- (N-methyl-Lvalyl) -L-prolyl)

  L-Prolyl-2-hydroxy-isovaleryl-2-oxo-4-methoxy-5 - [[(benzyloxycarbonylpropyloxy (methylphosphoryl) oxy) -phenyl] methyl] -3-pyrroline A solution of 64 mg of benzyl 4-methylphosphoryloxy-butanoate in 5 ml of dichloromethane are added 3 equivalents of triethylamine, 1 equivalent of the compound obtained in Step D of Example 1 and 3 equivalents of BOP (1H-benzotriazole-1 Hexafluorophosphate). yl-oxy) tris (dimethylamino) phosphonium). After 6 hours at 25 ° C., the solution is concentrated and then diluted in ethyl acetate, washed with aqueous solutions, dried, filtered and concentrated under reduced pressure. Chromatography on silica gel (dichloromethane / methanol

  / 5) to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z: 1107.

  Step B: (1S) -I - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-prolyl)

  L-Prolyl] -2-hydroxyisovaleryl-2-oxo-4-methoxy-S - [[(hydroxycarbonylpropyloxy (methylphosphoryl) -ox-yl] phenyl] methyl] -3-pyrroline A 90 mg of the compound of Stage A dissolved in 5 ml of tetrahydrofuran at 0 ° C is added 20% by weight of Pd / C and a flow of H2. After 1 hour, the solution is filtered

  and concentrated. Chromatography on silica gel makes it possible to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z: 1017.

  Stage C: Dol-Ph-O-Phos-But-OESi 7-OBz at -10 ° C., 8 mg of the compound of Stage B diluted in 0.5 ml of

  dichloromethane, 2.5 equivalents of N, N-diisopropyl-N-ethylamine. 1 equivalent of 3-

  benzoate-1-estradiol, 1 equivalent of DMAP and 2 equivalents of isopropenyl chlorocarbonate. After 4 hours at 25 ° C., the reaction is concentrated and chromatography on silica gel makes it possible to isolate the expected product.

(dichloromethane / methanol 95/5).

  Mass Spectrum: FAB +: [M + H]: m / z: 1375.

  High resolution: [M + Na +]: Calculated = 1397, 7014

found = 1398, 6329.

  o Example 6: Dol-Ph-O-Phos-But-OES17-OAII oM: o / -XMe oMe) o Me Me 0Me Me O / MO / 0 The procedure is as in Step C of Example 5 using as a substrate the compound

  obtained in Step A of Example 2 and the compound obtained in Step B of Example 5.

  Mass Spectrum: FAB +: [M + H + thioglycerol]: m / z: 1420.

  Example 7: Dol-Ph-O-Phos-But-OES17-OH, O, IO, Me

Me -

  The procedure is as in Example 3 using as substrate the compound obtained in

example 6.

  Mass Spectrum: FAB +: [M + H]: m / z: 1271.

  Example 8 Dol-Ph-O-Phos-But-NH-Et-OES11-OH

32 2774989

  ## STR2 ## to a solution of 3.5 mg of 11- (2'-aminoethyl) - ## STR2 ## Estradiol in 0.2 ml of dichloromethane is added 1 equivalent of triethylamine, then at 0 C 1.2 equivalents of the compound obtained in Step B of Example 5 and one equivalent of dicyclohexylcarbodiimide dissolved beforehand in dichloromethane. 25 C, the solution is concentrated under reduced pressure and chromatography on silica gel (90/10 dichloromethane / methanol)

allows to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z: 1314.

  EXAMPLE 9 DoI-Ph-O-Phos-But-NH-But-OESI1-OH OH O

/ 0-P-0 N X (C

  Me H0 0 Me, MeO N) s lf N. x) X <p N Hey Me O / \ Me O O ÈO

  The procedure is as in Example 8 using as substrate 11 - (4'aminobutyl) -

  estradiol and the compound obtained in Step B of Example 5.

  Mass Spectrum: FAB +: [M + H]: m / z: 1342.

  Example 10: Dol-Ph-O-Adi-NH-Et-OES1I-OH

O OH

O O

  Me H 0 Me NH (N-HO o t Me O / M MeO HA

O ,, y) O. ,, y. oi.

  Step A: (5S) -I- [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalvl) -L-prolyl)

  L-prolyl] -2-hydroxyisovaleryl] -2-oxo-4-methoxy-5 - [[4- (hydroxycarbonylbutylcarbonyloxy) -phenyl] methyl-3-pyrroline The procedure is as in Step A of Example 5 using as substrate the compound obtained in Step D of Example 1 which is reacted with monobenzyladipate, then the terminal carboxylic acid function is deprotected according to

  the same process described in Step B of Example 5.

  Mass Spectrum: FAB +: [M + H]: m / z: 981.

  Stage B: DoI-Ph-O-Adi-NH-Et-OES, OH To a solution of 4 mg of 11 (2- (2'-aminoethyl) estradiol hydrochloride in 0.5 ml of dichloromethane are added at 0.degree. C, 1 equivalent of triethylamine, 1 equivalent of the compound obtained in Step A and 1 equivalent of dicyclohexylcarbodiimide. After 4 hours at 25 ° C., the solution is concentrated under reduced pressure and chromatography on silica gel (dichloromethane / methanol

  95/5) makes it possible to isolate the expected product.

  Mass Spectrum: FAB +: [M + H]: m / z: 1278.

  Example 11: (5S) -1 - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-Lvalyl) -L-

  prolyl-L-prolyl] -2-hydroxyisovaleryl] -2-oxo-4- [2-amino- [17B] malonyloxy)

  (3-benzoyl-estradiol)] ethoxy] -5-benzyl-3-pyrroline = Do-O-Et-NH-MaI-OES17-Bz Me 0

N N N NO01

Me O Me O NH 0 0 Me OBz

  Stage A (5S) -i - [(2S) -O - [(N, N-dimethyl-L-valyl) -L-valyl- (N-methyl-L)

  valyl) -L-prolyl-L-prolvl] -2-hydroxy-isovaleryl] -2-oxo-4-aminoethoxy-5-

  benzyl-3-pyrroline The procedure is as in Step C of Example 1, using as substrate the compound obtained in Step 3 of Step A of Example 1, which is reacted with

  (5S) - 1 - [(2S) -2-Hydroxyisovaleryl] -2-oxo-4-benzyloxycarbonylaminoethoxy-5-benzyl-

  3-pyrroline, then hydrogenolysis according to the conventional method in methanol in the presence of Pd / C the product thus obtained.

Melting point: 138-142C.

  Mass Spectrum: FAB: [M + H]: m / z: 866.

  Stage B: Dol-O-Et-NH-Mal-OES] 7-Bz The procedure is as in Stage F of Example 1 using as substrate the

  compound of Step A and the compound of Step E of Example 1.

  Mass Spectrum: FAB +: [M + H]: m / z: 1310.

  High resolution: [M + H +]: theoretical = 1297, 7069

found = 1297, 7012.

  Example 12: Dol-O-Et-NH-Mal-OES17-AII "/ Hf o% / f _" NF Me O o A O

NN) N0

em, 0H NH O o.X'o o u

I

  The procedure is as in Step C of Example 5, using as substrate the compound

  obtained in Step A of Example 11 and the compound obtained in Step A of Example 2.

  Mass Spectrum: FAB +: [M + H]: m / z: 1246.

  EXAMPLE 13 ## STR6 ##

  The procedure is as in Example 3 using as substrate the compound of Example 12.

  Mass Spectrum: FAB +: [M + H] m / z: 1204.

  s Example 14: Bz-OES17-Mal-OThréo-Dol d H

O O

BMe M o0 Me 0OMe

N N

Me / \ Me o Bz

  Step A (55) -J - [(25) -O - [(N, N-dimethyl-L-threolyl) -L-valvl- (N-methyl-valvl) -L-

  prolyl-L-prolyl] -2-hydroxy-isovalérvl] -2-oxo-4-methoxy-5-Benzyl-3-

o pvrroline.

  The procedure is as in Step C of Example 1, using as substrate the compound obtained according to the same strategy as in Step A of Example 1, but replacing the last amino acid valine with a threonine amino acid. that is reacted with (5S) -1 - [(2S) -2-hydroxy-isovaleryl] -2-oxo-4-benzyloxycarbonyl 1-methoxy-5-benzyl-3-pyrroline, and then hydrogenolysis according to the conventional method, in

  methanol in the presence of Pd / C, the product thus obtained.

  Stage B: Bz-OES17-Mal-OThréo-Dol The process is carried out as in Stage F of Example 1 using the product obtained in

  Stage A which is reacted with the compound obtained in Step E of Example 1.

36 2774989

Example 15

  Me 1N. / VoMeO Me O O MeO S OMe O N // S

0 \ I

   The process is carried out as in Step C of Example 5 using as substrate Dolastatin 10 in which the carboxylic acid function on the thiazole ring is introduced according to the process described in the article J. Org. Chem. 1987. 52, 1252-1255, the compo

  intermediate obtained being reacted with oxalic acid.

  PHARMACOLOGICAL STUDY OF THE COMPOUNDS OF THE INVENTION

  Example 16: In Vitro Activity Murine leukemia L 1210 and human mammary adenocarcinoma MCF-7 were used in vitro. The cells are cultured in complete RPMI 1640 culture medium containing% fetal calf serum, 2 mM glutamine, 50 U / ml penicillin, 50 μg / ml streptomycin and 10 mM Hepes, pH 7.4 . The cells are distributed in microplates and exposed to the cytotoxic compounds for 4 doubling times, ie 48 hours for L1210 and 168 hours for MCF-7. The number of viable cells is then quantified by a colorimetric assay, the Microculture Tetrazolium Assay (Carmichael J. et al., Cancer Res., 47, 936-942, (1987).) The results are expressed as IC 50, cytotoxic concentration. who

  inhibits the proliferation of the treated cells by 50%.

  The compound of Example 4 thus has an IC50 of 24.1nM in the case of L 1210 and 1.5nM in the case of MCF-7, and the compound of Example 3 has an IC 50 of 6.8 nM in

37 2774989

  the case of L 1210 and 1.2 nM in the case of MCF-7. Dolastatin-15 presents

  respectively for these two lines of ICs0 of 0.24 nM and 0.19 nM.

  Example 17: Pharmaceutical composition: tablets Preparation formula for 1000 tablets dosed at 1 mg Composed of Example 3 .............

  ................................................ lg Lactose .............. DTD: .................................. ....................................... DTD: ..40 g Magnesium Stearate .. ........................................... DTD: ..... ............... 10 g Wheat starch .............................. .... DTD: ......................................... 15 g Starch of corn ............ DTD: .................................. ................... 15 g i0 Silica ........................... ......................................... .DTD: ...... .................. 3 g Hydroxypropylcellulose ...................... DTD: ..... ..................................... 5 g..DTD:

Claims (10)

1) Compounds of formula (I): (E) - (D) - (A) (I)   in which: (A) represents an oestradiol derivative, of formula: ## STR1 ## in which: R 1 represents a hydrogen atom, a linear or branched (C 1 -C 6) alkyl group, an alkenyl group (C 1 -C C6) linear or branched, a linear or branched (C1-C6) acyl group, an arylcarbonyl group or an arylalkylcarbonyl group in which the aryl part represents a phenyl group or a naphthyl group (these groups being optionally substituted by at least one atom of halogen, a linear or branched (C1-C6) alkyl group, linear or branched (C1-C6) alkoxy, linear or branched (C1-C6) acyl, cyano, nitro, or an amino group, itself optionally substituted with one or two linear or branched alkyl groups (C1-C6) and the linear or branched alkyl part contains from 1 to 6 carbon atoms, R2 represents a hydrogen atom, a linear or branched (C1-C6) alkyl group. an acyl group (C1-C6) lin or branched, an arylcarbonyl group or an arylalkylcarbonyl group having the same definition as in R1. or consists of a single bond (said bond corresponding to that existing between part (A) and part (D) of the compounds of formula (I)), R3 represents a hydrogen atom when R2 represents a single bond or R3 represents a single bond (said bond corresponding to that existing between the part (A) and the part (D) of the compounds of formula (1)) when R2 has one of the other definitions previously described, (D) represents a linear chain or branched, polysubstituted with formula -X Y Z-   in which X is connected at its free end to the part (E) of formula (I) and Z is connected at its free end to part (A) of formula (I) and where: X represents: - a group linear or branched (C1-C6) alkylene, O il - a carbonylalkylene group -CR- in which Ra represents a linear or branched (C1-C6) alkylene group, O il - or a group -POR- in which Ra represents a grouping linear or branched (C1-C6) alkylene and Rh represents a linear or branched (C1-C6) alkyl group, Y represents a carbonyl group or an amido group -NH-C- in which the carbonyl group is connected to the part Z of the group (D) of the formula (I), Z represents: - a single bond - an aminoalkylene -NH-Ra- group in which Ra represents a linear or branched (C1-C6) alkylene group, or an alkylene group carbonyl -Ra-C- in which Ra, has the same O definition as before, (E) feel: * a derivative of Dolastatine- 15 of the formula) R4 H O   Wherein: R4 represents: * is an isopropyl group and in this case: s Rs represents: - a group of formula (E1): [, v R6 -O N R 7 (E1)   in which: R 6 represents a hydrogen atom, or a linear or branched (C 1 -C 6) alkoxy group, and in this case R 7 represents an oxygen atom, said oxygen atom being connected to the group X of the part (D) of formula (I): where R6 represents an oxygen atom, said oxygen atom being connected to the group X of part (D) of formula (I), and in this case R7 represents a grouping linear or branched (C1-C6) alkoxy, or a group of formula (Ez): -NH- (CH2) -Rs (E2) in which Rs represents an oxygen atom, said oxygen atom being linked to the group X of the part (D) of the formula (I), n represents an integer between 1 and 4 inclusive, * is a group of formula CH 3 -CH-O- in which the oxygen atom is connected to the group X of the part (D) of the formula (I), and in this case: Rs represents: a group of formula (E '): R'6 -ON R'7 (E') or O   in which R6 'represents a hydrogen atom, or a linear or branched (C1-C6) alkoxy group, R'7 represents a linear or branched (C1-C6) alkoxy group, or a group of formula (E2') -NH- (CH 2) n -R '(E',) in which: R'g represents a hydrogen atom. or a linear or branched (C1-C6) alkoxy group, n represents an integer between 1 and 4 inclusive, * or a derivative of Dolastatin-10 of formula: M R4 H Me N N 9   Me O MeMe e 0 OMe O wherein: R4 represents: ; 422774989   * is an isopropyl group and in this case: R9 represents: - a group of formula (E.3): -N <> (E3)   Wherein R10 represents a group -C-O- (in which the oxygen atom is connected to the   to the group X of part (D) of formula (1)) or a group -CNH-   II O (in which the nitrogen atom is connected to the X group of part (D) of formula (I)) 1 ", - or a group of formula (E4): -NH- (CH2) n - R11 (E4) in which: R1 represents an oxygen atom, said oxygen atom being connected to the group X of the part (D) of the formula (I), n represents an integer between 1 and 4 inclusive, or a group of formula H CH-HO- in which the oxygen atom is connected to the group X of part (D) of formula (I) and in this case: R 9 represents: a group of formula (E'3): -N <(E'3) H R'N H R'10   in which: R'10 represents a hydrogen atom, - or a group of formula (E'4): -NH- (CHf), - R'l (E'4) in which: R'll represents a hydrogen atom or a linear or branched (C 1 -C 6) alkoxy group n represents an integer between 1 and 4 inclusive, their isomers, as well as their addition salts with an acid or a pharmaceutically acceptable base acceptable.   2) Compounds of formula (I) according to claim 1 characterized in that part (E) of formula (I) represents a derivative of Dolastatin-15 of formula as defined in claim 1, their isomers and their addition salts with an acid or a base pharmaceutically acceptable.   3) Compounds of formula (I) according to claim 1 characterized in that part (E) of formula (I) represents a derivative of dolastatin-10 of formula as defined in claim 1. their isomers and their addition salts with an acid or a base pharmaceutically acceptable.   4) Compounds of formula (I) according to any one of claims 1 to 3 characterized in that   that R4 represents an isopropyl substituent, their isomers as well as their addition salts to a   acid or a pharmaceutically acceptable base.   ) Compounds of formula (I) according to one of claims I or 2 characterized in that the   part (E) represents a derivative of Dolastatin-15 in which the group R 1 represents a group of formula (E 1) as defined above, their isomers as well as their   addition salts with a pharmaceutically acceptable acid or base.   6) Compounds of formula (I) according to one of claims 1 or 3 characterized in that the   part (E) represents a derivative of Dolastatin-10 in which the group R9 represents a group of formula (E3) as defined above. their isomers as well as their   addition salts with a pharmaceutically acceptable acid or base.   7) Compounds of formula (I) according to any one of claims 1, 2, 4 or 5 characterized   in that said compounds correspond to the formula (LA): \, - O-X-Y-Z-0 R4 0 0 me 7 - R3   N% N kNY (ÄI = II: _ N F JSN AO lN i M R O / \ Me O O f \ (lA) OR1   in which R 1 is as defined in formula (I) and preferably represents a hydrogen atom, a linear or branched (C 2 -C 6) alkenyl group, or an arylcarbonyl group, said group advantageously being a benzoyl group, R 3 represents a hydrogen atom, R4 represents an isopropyl group, R7 represents a linear or branched (C1-C6) alkoxy group, and preferably a methoxy group, OX represents a carbonylalkylene -CR- group. the carbonyl part being aa connected to the neighboring oxygen atom, or a group of formula - pO-Ra, Rb the phosphorus atom being connected to the neighboring oxygen atom, groups in which Ra represents an alkylene group (C1-C6) linear or branched and preferably of linear structure, and Rh represents a linear or branched (C1-C6) alkyl group, and particularly preferably, a methyl group, Y Y represents a carbonyl group -C -, and Z represents a single bond, their isomers and their addition salts with an acid or a pharmaceutically acceptable base acceptable.   8) Compounds of formula (I) according to any one of claims 1, 2, 4 or 5 characterized   in that said compounds correspond to the formula (IB): ## STR1 ## wherein R 1 is a compound of formula (II): ## STR2 ## in which: RI is as defined in formula (I) and preferably represents a hydrogen atom, a linear or branched (C2-C6) alkenyl group, or an arylcarbonyl group, said group being advantageously a benzoyl group, R3 represents a hydrogen atom, R4 represents an isopropyl group, R6 represents a hydrogen atom, X represents a linear or branched (C1-C6) alkylene group and preferably a linear (C1-C6) alkylene group, Y represents an amido -NH-C- group in which the -NH moiety is connected to the (X) moiety; Z represents an alkylene carbonyl -Ra-C- moiety in which the carbonyl moiety is attached to the atom neighbor oxygen, and Ra represents a grouping   linear or branched (C1-C6) alkylene, and preferably Ra is linear.   their isomers and their addition salts with an acid or a pharmaceutically acceptable base acceptable.   9) Compounds of formula (I) according to any one of claims 1, 2, 4 or 5 characterized   in that said compounds correspond to the formula (IC) OR \ ', O-X-Y-Z R R- H O O -   Embedded image in which R 1 is as defined in formula (I) and preferably represents a hydrogen atom, a (C 2 -C 6) alkenyl group; ) linear or branched, or an arylcarbonyl group. said group being advantageously a benzoyl group, R 2 is as defined in formula (I) and preferably represents a hydrogen atom or an arylcarbonyl group, said group advantageously being a benzoyl group, R 4 represents an isopropyl group, R 7 represents a linear or branched (C1-C6) alkoxy group and   preferential methoxy group.   X represents a carbonylalkylene group -CR--, the carbonyl part being a no longer connected to the neighboring oxygen atom, or a group of formula -pOR-, I a Rb the phosphorus atom being connected to the atom of adjacent oxygen, groups in which Ra represents a linear or branched (C1-C6) alkylene group and preferably Ra is linear, and Rb represents a linear or branched (C1-C6) alkyl group, and particularly preferably Rb represents a methyl group, Y represents a carbonyl group, Z represents an aminoalkylene -NH-Ra- group, the amine part being connected to Y and Ra representing a linear or branched (C1-C6) alkylene group, their isomers and their salts; addition to a pharmaceutically acceptable acid or base acceptable.   Process for the preparation of the compounds of formula (I) according to claim 1, characterized in that a compound of formula (II) is used as starting material: OR2 R'3 D ei) Ri in which R1 and R2 have the same definition as in formula (I) with the proviso that R2 is different from a single bond, and R'3 represents a hydrogen atom or a linear or branched (C1-C6) aminoalkyl group, * compounds of formula ( II) that, in the case where R 'and R2 simultaneously represent a hydrogen atom, in the presence of a coupling agent, a reaction is carried out in a basic condition with a compound of formula (III): 0 0   Wherein Ra is as defined in formula (I) and Rc is a protecting group of carboxylic acid functions,   to yield compounds of formula (IIa). O O Me ORc n J v (Ia) 48 2774989   in which R1. Ra and RK have the same definition as above, compounds of formula (ila), the terminal carboxylic acid function of which is deprotected under conventional conditions of deprotection of these functions to give compounds of formula (lib): O O O R, - 4OH   M '(IIb) R wherein R1 and Ra have the same definition as above, compounds of formula (IIb) which are coupled: * with a derivative of Dolastatin-15 of formula (IV / a): OHH O   I N 'x.J N N R5 NN N N R5 (IV / a) MeN / O o o--   Wherein R5 represents a group of formula (E'1) or (E'2) as defined in the general definition of formula (I), to yield compounds of formula (I / a), particular compounds of formula (I): O O Met_H O / M e N, N N- -I Rs N - N   Wherein R1 and Ra are as previously defined and R5 is as defined in formula (IV / a) or with a derivative of Dolastatin-15 of formula (IV / b). Embedded image in which R 7 represents a linear or branched (C 1 -C 6) alkoxy group, to give the compounds of formula (I) and (II). of formula (J / b), a particular case of the compounds of formula (1): O R O   Wherein R1 and Ra are as defined above and R7 is as defined in formula (IV / b). ## STR2 ## wherein R1 and Ra are as defined above and R7 is as defined in formula (IV / b). above, or with a derivative of Dolastatin-15 of formula (IV / c): Me NN, A%. N N NI- (CH) 1 OH N - N   In which n is an integer between 1 and 4 inclusive, to give compounds of formula (1 / c), a particular case of compounds of formula (I) ## STR2 ## (CH2). Embedded image in which n, R 1 and Ra are as defined above, or with a derivative of Dolastatin-10 of formula (V / a) OH Me y wherein R9 represents a group of formula (E'3) or (E'4) as defined in the general definition of the formula (## STR1 ## wherein I), to give compounds of formula (I / d), a particular case of compounds of formula (I): OO Me N N N N R9   Wherein R 1 and R 1 are as previously defined and R 9 is as defined in the formula (I). formula (V / a).   * with a derivative of Dolastatin-10 of formula (V / b): Me 0 M NN 'N N FNH- (CH2) OH   Wherein N is an integer from 1 to 4 inclusive, to yield compounds of formula (I / e). particular case of the compounds of formula (I): ## STR2 ## Ak À À:: OM Me Me Me (dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans dans; R1 and Ra are as defined above, * or with a compound of formula (VI): R6 Me III OR -NH, NN JNIs NW Me O \ me 0 o 0o (VI) in which Ra is as defined in the formula (I) and R6 represents a hydrogen atom or a linear or branched (C1-C6) alkoxy group, said compounds of formula (VI) being obtained by reaction, in the presence of triphenylphosphine and diethylazodicarboxylate, between a compound of formula (VI / a ): HO-R - NH-C-O-CH (VI / a)   in which Ra is as defined above.   and a compound of formula (IV / d) derived from Dolastatin-15: R6% H o / o Me OM N N N N O   Embedded image in which R6 represents a hydrogen atom or a linear or branched (C1-C6) alkoxy group, to give compounds of formula (VI / b) R6 Me NCA..N MN - Wherein Ra and R6 are as defined above, compounds of formula (VI / b), the function of which is as follows: ## STR5 ## wherein carbamate is reduced by hydrogenolysis in the presence of Pd / C, to give compounds of formula (VI) defined above, said compounds of formula (VI), brought into contact with said compounds of formula (IIb) previously described, in basic medium and in the presence of a coupling agent such as tris (dimethylamino) bromophosphonium hexatluorophosphate, lead to the compounds of formula (I / f), a particular case of the compounds of formula (I): yo /, / ". , X--, N o / R6 simultaneously with a hydrogen atom, X with a compound of formula (VII): O o o1 W-P-O-R OH (V)   Ra -Me, ORh in which Ra R6 and Rb are as defined in formula (I) and W is derived from a derivative of Dolastatin-15 or Dolastatin-10, telse (I that is reacted, in said cases where Rm and R2 simultaneously represent a hydrogen atom, X or with a compound of formula (VII): formula (VII) being obtained by addition in basic condition of a compound of formula (VIi / a): wherein R a and R b are as defined above and R represents a protecting group of carboxylic acid functional groups, such as a benzyl group, with one of the following groups: ## STR5 ## compounds of formula (IV / a), (IV / b), (IV / c), (V / a) and (V / b) at their free alcohol function, to give compounds of formula (VIIb), in which Ra, Rb and Rc are as defined above and W represents one of the residues of the compounds of formula (IV / a), (IV / b), (IV / c) ), (V / a) or (V / b) in which the atom e of hydrogen of the free alcohol function is substituted by the phosphorus atom of said compounds (VIlb), compounds of formula (Vllb) whose terminal carboxylic acid function is deprotected to give compounds of formula (VII), said compounds of formula (VII), brought into contact with said compounds of formula (I) under basic conditions and in the presence of isopropenyl chlorocarbonate, give compounds of formula (I / g), a particular case of compounds of formula (I): he '11 W-P-O - R O I _   Rh e (J / g) in which R 1, Ra and Rb are as defined above and W represents one of the residues of formula (IV / a), (IV / b), (IV / c), (V / a) and (V / b) in which the hydrogen atom of the alcohol function is substituted by the phosphorus atom of the compounds (I / g), X or with a derivative of Dolastatin-10 of formula (Vc) : Me H Me, to K NH   Me O Me OMeO OMe S NX o (Vc) N [M-Rummy O OH   in which Ra is as defined above and M represents an oxygen atom or a group -NH-, said compound of formula (Vc) being obtained by addition reaction, in basic condition and in the presence of a coupling reagent of a compound of formula (VIII): ## STR1 ## in which Ra and Re are as defined above and M 'represents a hydroxyl group or an amino group, with a derivative of dolastatin 10 of formula (Vd) Me 0 Me NMeO / M OMeO \ MeOMe O, SN Nu (Vd) OH to give compounds of formula (Ve) 1prcdMen td HOc Me N '-> - N N NHN   Embedded image in which Ra, R, and M are as defined above, compounds of formula (Ve), the terminal carboxylic acid function of which is deprotected under conventional conditions to give the compounds of formula (Vc) 1 5 previously described, said compounds of formula (Vc), brought into contact with said compounds of formula (II) under basic conditions and in the presence of isopropenyl chlorocarbonate, for example, lead to the compounds of formula (1 / h), a particular case of the compounds of formula (I): ## STR1 ## in which R1, Ra and M are as defined previously, s or compounds of formula (II) which are reacted, in the case where R 'represents a aminoalkyl group NH2Ra-   OR2 * or with a dRiv, of formula (VI are as defined above, in presence of a base such as triethylammon and a coupling agent, to give compounds of formula (I / i), particular case of Compounds of formula (I): O O OR2   Wherein R1, R2, Rm and Rb are as defined above and W represents one of the residues of formula (IV / a), (IV / b), (IV / c), (IV / c), V / a) and (V / b), the oxygen atom of the alcohol moiety is bonded to the phosphorus atom of the compounds (i / i),   or with a derivative of formula (IX). O o W-C-R OH (IX)   wherein Ra and W are as previously defined and said compounds of formula (IX) being obtained by addition reaction under basic conditions. and in the presence of a coupling agent, a compound of formula (IIl) as defined above with one of the compounds of formula (IV / a). (IV / b). (IV / c), (V / a) and (V / b) in their free alcohol function. to give compounds of formula (IXa): ## STR2 ##   wherein W represents one of the residues of the compounds of formula (IV / a). (IV / b). (IV / c).   (V / a) and (V / b) and Ra and RK are as defined above.   compounds of formula (DXa), the carboxylic acid function of which is deprotected under standard deprotection conditions in order to obtain the compounds of formula (IXa) as described. said compounds of formula (IX). brought into contact with said compounds of formula (II). in   basic condition in the presence of triethylamine for example, and a coupling reagent.   lead to the compounds of formula (1 / j), in particular the compounds of formula (I) ## STR2 ##   wherein R1, R2, Ra and W are as previously defined.   or with a derivative of formula (Vc) as defined above. to give compounds of formula (I / k), a particular case of compounds of formula (I): 57 2774989 H () M   vcN <Jk, N A N M -N0 m M, () _ bMai) Mc ((> Mc () t '( N. ') It OR. M -R. NH-R1   (1 / k) RiO in which R1, R2, Ra and M are as defined above, the compounds (I / a) to (I / k) forming all of the compounds of the invention, which are purified if appropriate, according to a conventional purification technique, which can, if desired, be separated into their different isomers according to a conventional separation technique, and which, if appropriate, are converted into their addition salts. to an acid or a base pharmaceutically acceptable.   11) Pharmaceutical compositions containing as active ingredient at least one compound   according to any one of claims 1 to 9, alone or in combination with one or more   non-toxic pharmaceutically acceptable inert excipients or carriers.   12) Pharmaceutical compositions according to claim 11 containing at least one principle   active agent according to any one of claims 1 to 9 useful in the treatment of tumors cancerous.