CA1198422A - N-(vinblastinoyl-23) derivatives of amino acids, their preparation and therapeutical application - Google Patents
N-(vinblastinoyl-23) derivatives of amino acids, their preparation and therapeutical applicationInfo
- Publication number
- CA1198422A CA1198422A CA000416996A CA416996A CA1198422A CA 1198422 A CA1198422 A CA 1198422A CA 000416996 A CA000416996 A CA 000416996A CA 416996 A CA416996 A CA 416996A CA 1198422 A CA1198422 A CA 1198422A
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- Canada
- Prior art keywords
- group
- alkyl
- hydrogen atom
- hydroxy
- methyl
- Prior art date
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
- C07D519/04—Dimeric indole alkaloids, e.g. vincaleucoblastine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
ABSTRACT OF THE DISCLOSURE
Vinblastine derivatives of the general formula:
(II)
Vinblastine derivatives of the general formula:
(II)
Description
4~
The presen-t invention relates to novel bisindole alkaloids. More par-ticularly, the invention relates to corn-bination products of amino aci.cl wi-th deriva-ti.ves of vin-blastine, -to a me-thod for -their preparation, tlleir use as anti-tumoral agents and to pharmaceutical compositions con -taining same~
These novel deriva-tives of vinblastine may be used in -treating leukemia and malignant tumors in humans.
Bisindole alkaloids of the vinblastine -type are well-known compounds having the carbon skeleton of the gen-eral formula:
Il 01 ~1130~C ~`~
The presen-t invention relates to novel bisindole alkaloids. More par-ticularly, the invention relates to corn-bination products of amino aci.cl wi-th deriva-ti.ves of vin-blastine, -to a me-thod for -their preparation, tlleir use as anti-tumoral agents and to pharmaceutical compositions con -taining same~
These novel deriva-tives of vinblastine may be used in -treating leukemia and malignant tumors in humans.
Bisindole alkaloids of the vinblastine -type are well-known compounds having the carbon skeleton of the gen-eral formula:
Il 01 ~1130~C ~`~
2~ C~
C~13 OR2 ' 23C~O
~1 .
As examples of -these compounds, one may cite:
vincaleukoblastine (U.S. Patent 3,097,137), leurocristine or vincristine and leurosidine (U.S. Paten-t 3,205,220), vingly-cinate ~Belgian Patent 659,112) and vindesine (Belgian Patent 813,168). The latter compound is ob-tained by chemical modifica-tion of natural vinblastine (I, R1 - OCH3, R2 =
COCH3), which is obtainable by extraction from Catharanthus ~t 4;~:
roseus leaves.
-Vinblastine, vincristine and vindesine are com-mercially available for use in human -therapy, more particu-larly for the treatmen-t of leukemia and some solid tumors.
However, these drugs have been shown to possess unfavourable side-effects. Vincristine shows neurotoxic , effects and vinblastine, a high toxici-ty as far as hemato-poetic tissues are concerned.
Their mechanism of ac-tion is similar -to the mech-anism which has been postula-ted for the antimitotic action of colchicine. I'hese drugs would act -through inhibition of the pol~merisation of tubuline to give micro-tubules, and subsequent arres-t of the cell division in the metaphase.
The u-tilization of 1:1 complexes of anti-tumoral tubiline-bisindole alkaloids has been described in Belgian Paten-t 854,053.
In some cases they show, lower toxici-t~ and ~ore efficient chemotherapeutic activi-ty than for the corresponding Eree alkaloids.
Other chemical modifications of vinblastine have been tested. Thus -the vinglycinate sulphate (I, sulphate, Rl = OCH3; R2 = COCH2 (CH3)2 -Cancer Research 1967, 27, 221-227), has also been tested in clinical experimen-tation but does not appear generally to be superior -to vinblastine or to vincristine.
Belgian Patent number 813,168 discloses Vindesine (I, Rl = NH~, R2 = H) and other vinblastine C3 carboxamide derivatives. Subsequent reports confirmed -the therapeutical interest of vindesine or 3-carboxamide 4-0-deacetyl vin-.;
~g~
blastine, bu-t this compound appears -to be inactive for the treatment murlne 1210 leukemia inocula-ted in mice (C.J.
Barnett et al., J. Med. Chem. 21, 88, 1978).
The new compounds described in the present inven-tion are able -to substantially delay the dea-th of mice in-travenously inoculated wi-th P 388 and L 1210 leukemias.
Ac-tivi-ties on experimen-tal P 388 tumors have ap-peared, surprisingly, very superior to reference Vinca alka-loids. Numerous total remissions have been observed.
The compounds oE the invention further show other important and unexpected advantages compared to vinblastine lS and known analogs, especially vindesine.
More particularly, the toxicity which has been observed is generally lower -than the corresponding toxicity of vindesine or vincristine.
The new compounds of the present inven-tion are more particularly those of the general formula IL:
C02CI~3' C~l3o ~2 ~5 o~ "
o
C~13 OR2 ' 23C~O
~1 .
As examples of -these compounds, one may cite:
vincaleukoblastine (U.S. Patent 3,097,137), leurocristine or vincristine and leurosidine (U.S. Paten-t 3,205,220), vingly-cinate ~Belgian Patent 659,112) and vindesine (Belgian Patent 813,168). The latter compound is ob-tained by chemical modifica-tion of natural vinblastine (I, R1 - OCH3, R2 =
COCH3), which is obtainable by extraction from Catharanthus ~t 4;~:
roseus leaves.
-Vinblastine, vincristine and vindesine are com-mercially available for use in human -therapy, more particu-larly for the treatmen-t of leukemia and some solid tumors.
However, these drugs have been shown to possess unfavourable side-effects. Vincristine shows neurotoxic , effects and vinblastine, a high toxici-ty as far as hemato-poetic tissues are concerned.
Their mechanism of ac-tion is similar -to the mech-anism which has been postula-ted for the antimitotic action of colchicine. I'hese drugs would act -through inhibition of the pol~merisation of tubuline to give micro-tubules, and subsequent arres-t of the cell division in the metaphase.
The u-tilization of 1:1 complexes of anti-tumoral tubiline-bisindole alkaloids has been described in Belgian Paten-t 854,053.
In some cases they show, lower toxici-t~ and ~ore efficient chemotherapeutic activi-ty than for the corresponding Eree alkaloids.
Other chemical modifications of vinblastine have been tested. Thus -the vinglycinate sulphate (I, sulphate, Rl = OCH3; R2 = COCH2 (CH3)2 -Cancer Research 1967, 27, 221-227), has also been tested in clinical experimen-tation but does not appear generally to be superior -to vinblastine or to vincristine.
Belgian Patent number 813,168 discloses Vindesine (I, Rl = NH~, R2 = H) and other vinblastine C3 carboxamide derivatives. Subsequent reports confirmed -the therapeutical interest of vindesine or 3-carboxamide 4-0-deacetyl vin-.;
~g~
blastine, bu-t this compound appears -to be inactive for the treatment murlne 1210 leukemia inocula-ted in mice (C.J.
Barnett et al., J. Med. Chem. 21, 88, 1978).
The new compounds described in the present inven-tion are able -to substantially delay the dea-th of mice in-travenously inoculated wi-th P 388 and L 1210 leukemias.
Ac-tivi-ties on experimen-tal P 388 tumors have ap-peared, surprisingly, very superior to reference Vinca alka-loids. Numerous total remissions have been observed.
The compounds oE the invention further show other important and unexpected advantages compared to vinblastine lS and known analogs, especially vindesine.
More particularly, the toxicity which has been observed is generally lower -than the corresponding toxicity of vindesine or vincristine.
The new compounds of the present inven-tion are more particularly those of the general formula IL:
C02CI~3' C~l3o ~2 ~5 o~ "
o
- 3 !!
-wherein: R is an es-ter of an amino-acid which is possibly protected, attached to the vinblastin-23-oyl moiety by an amide bond, the ester group, which may be s-traight or bran-ched, con-taining 2-9 carbon atoms and Rl is a hydrogen atom or a hydroxy group, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R5 is a hydrogen atom or a methyl group or a formyl group, with the provision that compounds where, simultaneously, R5 is methyl and Rl is a group ~-hydroxy ' are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids.
The Eollowing compounds are particularly preferred:
I R
CO. ('11 ' 3' (III) C113~ 1 1 2 R C~ C~
o where Rl is a hydroxy group or a hydrogen a-tom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched Cl-C8 alkyl, hydroxy-Cl-C8-alkyl, aminohydroxy-Cl-C8-alkyl, carboxy-Cl C8-alkyl, amido-Cl-C8-alkyl, guanadino-Cl-C8-alkyl, amino-Cl-C8-alkyl, thiol-- -
-wherein: R is an es-ter of an amino-acid which is possibly protected, attached to the vinblastin-23-oyl moiety by an amide bond, the ester group, which may be s-traight or bran-ched, con-taining 2-9 carbon atoms and Rl is a hydrogen atom or a hydroxy group, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R5 is a hydrogen atom or a methyl group or a formyl group, with the provision that compounds where, simultaneously, R5 is methyl and Rl is a group ~-hydroxy ' are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids.
The Eollowing compounds are particularly preferred:
I R
CO. ('11 ' 3' (III) C113~ 1 1 2 R C~ C~
o where Rl is a hydroxy group or a hydrogen a-tom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched Cl-C8 alkyl, hydroxy-Cl-C8-alkyl, aminohydroxy-Cl-C8-alkyl, carboxy-Cl C8-alkyl, amido-Cl-C8-alkyl, guanadino-Cl-C8-alkyl, amino-Cl-C8-alkyl, thiol-- -
4~;
Cl-C8-alkyl, cysteinyl-methyl, me-thylthio-e-thyl, benzyl, hydroxy-benzyl, or a group:
, ( ) C112~
or R3 together with the carbon -to which i-t :is attached and the ni-trogen of ~he amido-group, forms an azole or an hydroxy-azole ring; R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched Cl-C8-alkyl, or a benzyl group, with the provision tha-t compounds where, simultaneously, Rl is ~-OH and R5 is -CH3 are ex-cluded, -COOR4 is preferably the carboethoxy or -the carbome-thoxy groupj or their addition salts with a mineral or organic acid.
In a preferred embodiment, the structural segment R13 1l in general formula TII, represents an ester derived from any of the naturally-occuring amino acids and their optical isomers of D-configuration, namely glycine, alanine, valine, leucine, isoeucine, serine, -threonine, aspartic acid, gluta-mic acid, aspargine, glu-tamine, arginine, lysine, cys-teine, cystine, methionine, phenylalanine, tyrosine, -tryptophane, proline, histidine hydroxy-lysine or hydroxy-proline.
The compounds may be designated as 3-decarbomethoxy-0-4-deacetyl vinblas-tine-3-carboxamine. They will however be preferably designated as N-(vinblas-tin-23-oyl) or N-(vincris-tinoyl-23) deriva-tives of amino-acids hereafter.
Par-ticularly preferred compounds of the invention of general formula II are -those wherein R2 is hydrogen and the amino acid moie-ties are derived from one of the following
Cl-C8-alkyl, cysteinyl-methyl, me-thylthio-e-thyl, benzyl, hydroxy-benzyl, or a group:
, ( ) C112~
or R3 together with the carbon -to which i-t :is attached and the ni-trogen of ~he amido-group, forms an azole or an hydroxy-azole ring; R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched Cl-C8-alkyl, or a benzyl group, with the provision tha-t compounds where, simultaneously, Rl is ~-OH and R5 is -CH3 are ex-cluded, -COOR4 is preferably the carboethoxy or -the carbome-thoxy groupj or their addition salts with a mineral or organic acid.
In a preferred embodiment, the structural segment R13 1l in general formula TII, represents an ester derived from any of the naturally-occuring amino acids and their optical isomers of D-configuration, namely glycine, alanine, valine, leucine, isoeucine, serine, -threonine, aspartic acid, gluta-mic acid, aspargine, glu-tamine, arginine, lysine, cys-teine, cystine, methionine, phenylalanine, tyrosine, -tryptophane, proline, histidine hydroxy-lysine or hydroxy-proline.
The compounds may be designated as 3-decarbomethoxy-0-4-deacetyl vinblas-tine-3-carboxamine. They will however be preferably designated as N-(vinblas-tin-23-oyl) or N-(vincris-tinoyl-23) deriva-tives of amino-acids hereafter.
Par-ticularly preferred compounds of the invention of general formula II are -those wherein R2 is hydrogen and the amino acid moie-ties are derived from one of the following
- 5 -~:~9~
amino-acids: L or D -tryptophan, leucine, isoleucine, valine, alanine or phenyl-alan:ine. ~mong these, L,-txyptcphane, L-isoleucine and L-alanine are particularly interesting.
The present invention describes more particularly reac-tion produc-ts of amino-acids with vincristine, 0-4-deacetyl vincristine, deformyl-0-4-deacetyl vincristine and deoxy-vinblastine, ~.
The latter compound may be obtained by hydrogena--tion of anhydrovinhlatine obtained by dehydratation of vin-blasine or by coupling vindoline with catharantine (Po-tier et al, JACS 98 7017, ~1976).
_ Most particularly preferred compound of the presen-t inven-tion is ethyl N-(0-4-deacetyl-4'-deoxyvinblastin-23-ovl)-tryptophante.
When the amino acid contains functional groups in the side chain R3 (formula III) such as lysine or cysteine i-t may be necessary to protect the functional groups accord-ing to methods well-known to those skilled in the art.
Pro-tection may be achieved, depending on -the nature of the gxoup -to be protec-ted by condensing a benzyl, t-butyl, benzyloxycarbonyl, t--bu-toxy--triEluoroacetyl carbonyl or trityl radical. Other well-known protecting groups in use in pep-tide chemistry may successfully be used.
The compounds of the present inven-tion may be pre-pared in the conventional manner starting from corresponding derivatives of vinblastine, by hydrozinolyse followed by nitrosation and reaction with the amino acid ester in accor-dance with the reaction sequence I
~:~9~Z~
N211,~ Ncl1`102 ~/b Cll Oll I llCl < ~r7 OCOC113 3 ~/~ 0C ~< ~OII
COOCll~ 110 C-N~ 11-12 110 C~ 3 O O
'ester of amillo- -~cid C112C12 J-- l OII
~lo c-r~-c~co-o~, l~eaction sequence I
The first s-tep of the process is -to add anhydrous 0 hydrazine inexcess to a solution of vinblas-tine base in anhydrous methanol. The solution is heated in an inert atmosphere for 12 hours to 12 days a-t a temperature be-tween about 30C and 70C. Mos-t preferably the temperature is maintained around 60C.
The hydrazide of the bis-indole derivative is then isolated by adding water, extracting with dichlorome-thane and concentrating. The cornpound may be later purified by 30 preparative chromatography (neu-tral silica). In -the case of vincristine, the product obtained is the 0,4-deace-tyl-deformyl-vincristne carboxyhydrazide.
During the second step, the hydrazide function of -the modified vinblastine is transformed in-to the acyl azide.
This transformation is achieved in a known manner by adding 9~34Z~
sodium nitrite to the hydrazide dissolved in a water-methanol-acidic mixture.
The acid which is used may be hydrochloric acid.
The reaction tempera-ture is main-tained between 0C
and 5C. AEter extraction with a water-immiscible apro-tic solvent, prefera,bly methylene chloride, -the organic phase , is partially concen-trated.
The acyl azide proper is preferably not isolated bu-t directly added to the amino acid es-ter or possibly a protec-ted derivative thereof, dissolved in me-thylene chloride.
The quan-ti-ty of amino acid, -to be used is abou-t one to five equivalents of the modified vinblastine car-boxazide.
The reaction mixture is main-tained between abou-t -3C and ~10C for 8-72 hours and generally for about 15 hours. Monitoring of the reaction is easily achieved by thin layer chromatography. Af-ter concentrating, the compound of -the invention of formula II may be -transformed into a sulphate salt or ano-ther salt derived from a mineral or organic acid, by crystallization from a methanolic solution of the corresponding acid.
The compounds of the invention may be purified by conventional techniques of chromatography and re-crystalli-zation.
If desired, the thus obtained ~-0-deacetyl vin-blastine derivative can be reacylated ei-ther directly -to give the vinblastine deriva-tive III wherein R2 is COCH3 (~. Med.Chem.22, 391, 1979) or -through the preliminary forma-tion of the 3,4-diacetoxy derivative followed by a ~j - 8 -~1 '.
selective hydrolysis oE the 3-acetoxy group in the position 3. rhe hydroxy group :in C4 may be, in a conventional manner, esterified by other ac-tiva-ted acid deriva-tives containing 2-9 C atoms. The 0-4 formyl derivatives are obtained by formyla-tion (formic acid-acetic anhydrid) of the correspon-ding 0--4 deacetyl compounds (see Belgian Patent 660,843).
The present invention rela-tes also to the indus-trial and par-ticularly pharmaceutical applications of the new bisindole alkaloids.
The compounds of this invention display particularly very useful antitumor properties which may be applied in human therapy.
These amino-acid deriva-tives may be used for the treatmen-t of L 1210, P 388 type leukemia, gliomas, lympho-sarcomas and o-ther leukemias or malignant tumors. In human medicine they are used for the -treatment of Hodgkin's disease and for other solid -tumors treatable with vinblastine, vincristine or vindesine. These compounds arealso useful in veterinary medicine for the treatment of tumors of the animals.
O-ther in-teresting therapeutical uses may also be contempla-ted for the new derivatives of bisindole alkaloids.
[t has been descr:ibed tha-t vinblastine may be used for treat-ing some forms of arthritis (U.S. Patent 4,208,411) and -that vincristine may be used for treating psoriasis (U.S. Patent 3,749,984).
For their therapeu-tical use, -the compounds of the invention, possibly in the lyophilized form, are preferably adminis-tered by parenteral route, dissolved in a pharma-ceu-tically acceptable solvent, in the form of a base or of a pharmaceutically acceptable acid addition salt. Among the ~, ' 1~ 9~ 2~:
acid addition salts, the non-toxic and pharmaceutically acceptable sa:Lts, such as inorganic acid salts as hydro-chloric acid, phosphoric acid and sulurie acid salts or organic acid sal-ts as acetic, propionic, succinic, -tartrie, oxalic, methansulEonic and benzene-sulfonic aeid sal-ts are preferred.
Physiological water and other saline solutions , which have been buffered, for instance with a phosphate are appropriate solvents. In general, the compounds can be used in human therapy in an analogous manner to the technique and limitations in use for other alkaloids of -the Vinca type.
The active substance is generally distributed at a posology varying between 0.01 mg to about 5 mg/kg depend-ing upon the derivative and the therapeutic schedule which is adopted. Total weekly doses will vary generally between 0.1 and about 35 mg/kg.
The compositiorls in accordance with the inven-tion are prepared as unitary doses of 2 to 900 mg.
The compounds of the invention may be used alone or in combination with other carcinosta-tic agents including, for exampler -the alkylating agents, the an-ti-metabolites such as methotrexate, 5-fluoro-uracil, 6-mercap-topurine, 6-thio-guanine, -the cystosine arabinosides, and antibiotics such as actinomycine D, daunorubicine, adriamycine and cis-diamo-dichloro-platine. In particular, the new vinblastine derivatives may be used in combination.
The following specific examples illustrate in a non-limitative way the process for obtaining the compounds of the invention.
,4Z~
Example_ 1. Prepara-tion of 0--4-deacetyl N-deformyl-vincristine mono-hydrazide A.
985 mg of vincris-tine base are dissolved in a mix-ture of 10 ml anhydrous hydrazine and 10 ml methanol. The reaction mixture is mainta,ined a-t 60C, while stirring, for 22 hours.
' The reaction mixture is -then treated wi-th water and -there-af-ter with wa-ter sa-turated by NaCl. The aqeuous solution is extracted 8 times with 15 ml dichloromethane. The collec-ted organic phases are treated with 20 ml water, -thereafter 25 ml water saturated by NaCl and finally dried on Na2SO4.
After filtration and vacuum concen-tration, 900 mg of 0-4-deacetyl vincristine monohydrazide (purity over 90%) have been obtained.
NMR spectrum (CDC 13,360 MHz) 9.76 (s,lH), 8.37 (s,lH), 8.06 (s,lH), 7.53 (d,lH), 7.24 and 7.07 (m,3H), 6.82 (s,lH), 6.22 (s,l), 5.87 (dd,lh), 5.67 (d,lH), 5.43 ~s,lH), 4.03 (s,lH), 3.94 (s,3H), 3.86 (s,lH), 3.73 (s,3H), 3.52 (s,3H), 2.82 (s,2H), 2.51 (s,lH), 0.93 (2t, 2 x 3H).
Mass Spectrum (elec-tronic ionisa-tion impact) 154, 294, 355, 413, 524, 555, 695, 710, 723, 736, 739, 755 (M 1) - 768, 769, 782 calculated for C42H54N6O7 754 2. Prepara-tion o~ acid azide of modified vincristine A solu-tion of 850 mg of vincristine monohydrazide A in 20 ml methanol is trea-ted with 63 ml HCl 1 N and 175.5 mg sodium nitrite for 15 lminutes at 0C.
;
I'he solution maintained at 0C is then neu-tralised by NaHCO3 at 0C and thereaEter extracted 3 times with dichloromethane.
The collected organic phases are dryed on Na2SO4, filtrated and vacuum concentrated without heating until a solution of about 10 ml is ob-tained.
3. Reaction with ethyl -tryptophana-te.
300 mg of ethyl -tryptophanate are then added while stirring at about 4C and the reaction is checked by tlc. After 10 days, the reaction product is purified by chromatographic separation (40 g silicagel 60, elution solvent ether /NH3 saturated methanol 96:4 volume/volume)~ Fractions of 15 ml are collected which are checked by -tlc. The non-reacted amino-acid is first collec-ted (fraction 30 to 40). The elu-tion agent is changed (ether/NH3 sa-turated methanol 86:14) and -the reacted deriva-tives are collected as -three frac-tions:
fractions 44 and 45:30 m~3; fractions 46-53:257 mg; fraction 54-60:115 mg).
The second part was ethyl N-(0-4-deace-tyl deformyl vincris-tine-23-oyl) tryptophanate Ia which was homogeneous by tlc.
Mass spectru : 984, 970, 956 (M -~ 1) 913, 912, 899 (DCJ
isobu-tane), calculated for C55H66N609 : 955,181.
~V spectrum (CH OH) :
max: 282 (4.20) - 290 (4.18) - 322 (3.96) min: 254 - 287 - 305 IR spectrum: (cm :
_MR Spec-trum (CDC13) 360 MH (ppm) 8.50 (s,lH ),8.03 (s,2H), 7.77 (d,lH), 7.60 (d,lH), 7.57 (d, lH), 6.95 (s,lH), 6.35 (s,lH), 5.89 (d.d.,lH), 5.78 (d,lH), 4.75 (q,lH), 3.88 (s,3H), 3.67 (s,3H), 1.23 (t,3H), 0.98 (t,3il), 0.89 (-t,3H) Ethyl N-(0,4-deacetyl-vincristine-23-oyl)-tryp-tophanate IB.
, The deformylated derivative I a (257 mg) is re-formylated by applying the method described in Belgian Pa-tent 811,110.
The raw product -thus obtained is purified by chromatographic separation using a 20 cm column (silicagel, 30 g) and succe-sively 3 elu-tion agen-ts: ether/NH3 satura-ted MeOH 92:8;
88:12, 86:14. The effluent is collected as 15 ml fractions.
The fractions 44-54 (34.1 mg) contained the N-reformylated derivative Ib.
Mass spectrum (isobu-tane DCI) : 1011,997, 983 (M + 1), 970, 982, 996, 998, 999, 1012 calculated for C56H66N6010 = 983,192 UV spectrum (CH30H) max: 270 (4.18) - 290 (4.11) plate 280 (4.12) shouldering (4.03) IR _pec-trum (KBr) bands at 3400 - 3040 - 2960 - 2920 - 2880 - 2850 - 1735 -1725 - 1680 - 1670 1665 - 1610 - 1490 - 1450 - 1225 - 7i~5.
Example 2 Ethyl N-(0-4 deacetyl-deoxy-4'-vinblas-tine-23-oyl-B) -tryphtophante Ic.
~9~4~Z
The 0-4-deacetyl-deoxy-vinblastine hydrazide is prepared by the me-thod described in U.S. Pa-tent 4,203,898 (assigned to Ely Lilly Co., column 24, line 51 and following).
500 mg of the hydrazide in 12 ml methanol are -thereafter treated wi-th 37 ml HC1 lN and 104 mg NaNO2 for 15 minutes at 0C
The solu-tion is -thereafter neutralized by NaHCO3 and extrac-ted six times with 15 ml dichloromethane. The organic phases are dryed on Na2SO4 and concentrated until a volume of about 10 ml is obtained.
The azide in solution is then brought into con-tact with ethyl tryptophanate (300 mg) at 4C while s-tirring. The reaction is checked by tlc (ether, MeOH). Af-ter 6 days, the reaction is comple-ted and the reaction produc-t is purified by column chroma-tography on silicagel (18 cm, 30 g), elution by 15 ml Eractions.
The following elution agents have been successively used:
E-ther 96 NH3 sat, MeOH 4 frac-tions 1-49 92 8 frac-tions 50-57 86 14 fractions 71-90 The fractions 50-57 contain the wanted reac-tion product Ic (108 mg).
The sulphate is prepared by e-ther precipi-tation (2% H2SO
ethanol).
W spectrum of Sulphate (CH30H) max: 224 (4.63) - 272 (4.32) min: 247 (4.03) , 34~2 shouldering: 286 (4,22) - 312 (3,72) IR spec-trum (KBr) of Sulphate 3420 - 3060 - 2960 - 2930 - 2880 - 2600 (weak) - 1725 -1060 - 1015 - 920 - 745 cm 1, NMR spectrum (CHCl ) MHz of the base in ppm.
9.46 s lH 7.53 d lH 4.94 q lH
8.34 s lH 7.30 d lH 4.18 d lH
7.92 s lH 6.51 s lH 4.05 q 2H
7.67 d 1ll 6.04 s lH 3.77 s 3H
7.59 d lH m. about s. centered on 5.81 - 3.58 s RH
3.47 s lH
2.82 s 2H
2.77 s 3H
2.59 s lH
1.14 t 3H
0.95 t 3H
0.87 t 3H
Mass spectrum of the base DCI isobutane M + 1 at 954 M + 14 + 1 at 968 M + 28 + 1 at 982 ions at 153 - 155 - 157 - 171 - 172 - 185 186 - 199 -calculated for C56 H68 N6 8 Example 3 Etilyl N-(O-4-~eace-tyl-4'-deoxy-vinblas-tin-23-oyl-B so leucinate.
A solution of O-4~deacetyl-4'-deoxyvinblastine hydrazide (0.6, 0.79 mM - see U.S. patent 4,203,898 column 24, line 51) in a mi~-ture of 15 ml of CH30H and 45 ml HCl l N
, was coo]ed to -7~C. NaNO2 was added to the solution (0.15 g) and stirring was continued for 13 minutes a-t -7C. After addi-tion of an aqueous saturated solution of NaHCO3 -till a pH-value of 9 is obtained, -the resulting solu-tion is ex-tracted four times with CH2C12. The combined organic phases were washed wi-th sat.aqueous NaCl solution and dried over ~5 MgSo4.
The solution was concentrated to a volume of 15 ml and ethyl isoleucinate (1.25 mM) was added. The reaction mixture was allowed -to stand 4 days in a refrigerator. The solvent was then evaporated under reduced pressure. The residue was purified by column chromatography (SiO2, 60 g) and eluted with ether/CFl3OH/NH3 (96:4)Ø25 g of amorphous produc-t (yield 35~) have been thus obtained.
Mass spectrum: (Molec.Ionisation isobutane) 880 (M ), 894, 836, 849, 832, 445; 391, 355 cm Melting poin-t 180-190 C
~D = 104 (c=0.154) IR (CHC13) 3470, 2970, 1728, 1655, 1612, 1502 cm UV (CH30H) 289, 268, 216 nm NMR ~CDC13) 7.9 (NH), 6.54 and 6.06 Hg, H12, 5.81 (H14-H15) 4.06 (CHCO2(NH)) 3.76 CO2CH3, 3.58 CH3-O, 3.33 and 2.85 (H3A and H3B), 2.73 (N-CH3), 0.69 (H-15')-!~
~9~22 Acute toxicity-Determination of LDSo The acute -toxicity of the compound of Example 2:
ethyl N-(O-4-deace-tyl-deoxy-4'-vinblastine-23-oyl-B) try-phtophanate has been determined on female NMRI mice. In-creasing doeses of -the drug have been inoculated intravenously in a single injection. The mortality percentage is deter--mined as a functi,on of -the time. The LD50 values for -the ' above compound (deoxy VTrpE) and a reference product: deoxy-vinblastine (deoxy VBL) are indicated hereafter Drug LD50 Deoxy VBL 20 Deoxy VTrpE 91.5 (i.e. Compound The compound deoxy VTrpE is thus less toxic -than the reference compound.
Antitumoral activity The chemiotherapuetic activity of the sulfa-te salt of deoxy VTrpE has been studied on the lymphoblas-tic leu-kemia L 1210 and P 388.
emale DBA2 mice have been inoculated by 104 leu-kemic cells. The produc-t is administered by intravenous route, the day after the graft at a 50mg/kg dose. The mor-tality of the animals is observed as a function of time.
Table I hereafter indica-tes the results obtained wi-th com-pound of Example 2 (Ic). At the tested dose, -the compound is more active than vinblastine. This superiority has been confirmed compared to deoxy-vinblastine.
Female DBA2 mice are in-travenously inocu:lated by 10 leukemic cells. The pro~uc-ts are i.v. adminis-tered, -the day after the graf-t. I'he mortality of -the animals is observed as a func-tion of -time. Resul-ts appear in Table I.
Deoxy VTrpE is clearly more active than the reference com-pound and indica-tes a higher percen-tage of surviving subjects , at long term.
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amino-acids: L or D -tryptophan, leucine, isoleucine, valine, alanine or phenyl-alan:ine. ~mong these, L,-txyptcphane, L-isoleucine and L-alanine are particularly interesting.
The present invention describes more particularly reac-tion produc-ts of amino-acids with vincristine, 0-4-deacetyl vincristine, deformyl-0-4-deacetyl vincristine and deoxy-vinblastine, ~.
The latter compound may be obtained by hydrogena--tion of anhydrovinhlatine obtained by dehydratation of vin-blasine or by coupling vindoline with catharantine (Po-tier et al, JACS 98 7017, ~1976).
_ Most particularly preferred compound of the presen-t inven-tion is ethyl N-(0-4-deacetyl-4'-deoxyvinblastin-23-ovl)-tryptophante.
When the amino acid contains functional groups in the side chain R3 (formula III) such as lysine or cysteine i-t may be necessary to protect the functional groups accord-ing to methods well-known to those skilled in the art.
Pro-tection may be achieved, depending on -the nature of the gxoup -to be protec-ted by condensing a benzyl, t-butyl, benzyloxycarbonyl, t--bu-toxy--triEluoroacetyl carbonyl or trityl radical. Other well-known protecting groups in use in pep-tide chemistry may successfully be used.
The compounds of the present inven-tion may be pre-pared in the conventional manner starting from corresponding derivatives of vinblastine, by hydrozinolyse followed by nitrosation and reaction with the amino acid ester in accor-dance with the reaction sequence I
~:~9~Z~
N211,~ Ncl1`102 ~/b Cll Oll I llCl < ~r7 OCOC113 3 ~/~ 0C ~< ~OII
COOCll~ 110 C-N~ 11-12 110 C~ 3 O O
'ester of amillo- -~cid C112C12 J-- l OII
~lo c-r~-c~co-o~, l~eaction sequence I
The first s-tep of the process is -to add anhydrous 0 hydrazine inexcess to a solution of vinblas-tine base in anhydrous methanol. The solution is heated in an inert atmosphere for 12 hours to 12 days a-t a temperature be-tween about 30C and 70C. Mos-t preferably the temperature is maintained around 60C.
The hydrazide of the bis-indole derivative is then isolated by adding water, extracting with dichlorome-thane and concentrating. The cornpound may be later purified by 30 preparative chromatography (neu-tral silica). In -the case of vincristine, the product obtained is the 0,4-deace-tyl-deformyl-vincristne carboxyhydrazide.
During the second step, the hydrazide function of -the modified vinblastine is transformed in-to the acyl azide.
This transformation is achieved in a known manner by adding 9~34Z~
sodium nitrite to the hydrazide dissolved in a water-methanol-acidic mixture.
The acid which is used may be hydrochloric acid.
The reaction tempera-ture is main-tained between 0C
and 5C. AEter extraction with a water-immiscible apro-tic solvent, prefera,bly methylene chloride, -the organic phase , is partially concen-trated.
The acyl azide proper is preferably not isolated bu-t directly added to the amino acid es-ter or possibly a protec-ted derivative thereof, dissolved in me-thylene chloride.
The quan-ti-ty of amino acid, -to be used is abou-t one to five equivalents of the modified vinblastine car-boxazide.
The reaction mixture is main-tained between abou-t -3C and ~10C for 8-72 hours and generally for about 15 hours. Monitoring of the reaction is easily achieved by thin layer chromatography. Af-ter concentrating, the compound of -the invention of formula II may be -transformed into a sulphate salt or ano-ther salt derived from a mineral or organic acid, by crystallization from a methanolic solution of the corresponding acid.
The compounds of the invention may be purified by conventional techniques of chromatography and re-crystalli-zation.
If desired, the thus obtained ~-0-deacetyl vin-blastine derivative can be reacylated ei-ther directly -to give the vinblastine deriva-tive III wherein R2 is COCH3 (~. Med.Chem.22, 391, 1979) or -through the preliminary forma-tion of the 3,4-diacetoxy derivative followed by a ~j - 8 -~1 '.
selective hydrolysis oE the 3-acetoxy group in the position 3. rhe hydroxy group :in C4 may be, in a conventional manner, esterified by other ac-tiva-ted acid deriva-tives containing 2-9 C atoms. The 0-4 formyl derivatives are obtained by formyla-tion (formic acid-acetic anhydrid) of the correspon-ding 0--4 deacetyl compounds (see Belgian Patent 660,843).
The present invention rela-tes also to the indus-trial and par-ticularly pharmaceutical applications of the new bisindole alkaloids.
The compounds of this invention display particularly very useful antitumor properties which may be applied in human therapy.
These amino-acid deriva-tives may be used for the treatmen-t of L 1210, P 388 type leukemia, gliomas, lympho-sarcomas and o-ther leukemias or malignant tumors. In human medicine they are used for the -treatment of Hodgkin's disease and for other solid -tumors treatable with vinblastine, vincristine or vindesine. These compounds arealso useful in veterinary medicine for the treatment of tumors of the animals.
O-ther in-teresting therapeutical uses may also be contempla-ted for the new derivatives of bisindole alkaloids.
[t has been descr:ibed tha-t vinblastine may be used for treat-ing some forms of arthritis (U.S. Patent 4,208,411) and -that vincristine may be used for treating psoriasis (U.S. Patent 3,749,984).
For their therapeu-tical use, -the compounds of the invention, possibly in the lyophilized form, are preferably adminis-tered by parenteral route, dissolved in a pharma-ceu-tically acceptable solvent, in the form of a base or of a pharmaceutically acceptable acid addition salt. Among the ~, ' 1~ 9~ 2~:
acid addition salts, the non-toxic and pharmaceutically acceptable sa:Lts, such as inorganic acid salts as hydro-chloric acid, phosphoric acid and sulurie acid salts or organic acid sal-ts as acetic, propionic, succinic, -tartrie, oxalic, methansulEonic and benzene-sulfonic aeid sal-ts are preferred.
Physiological water and other saline solutions , which have been buffered, for instance with a phosphate are appropriate solvents. In general, the compounds can be used in human therapy in an analogous manner to the technique and limitations in use for other alkaloids of -the Vinca type.
The active substance is generally distributed at a posology varying between 0.01 mg to about 5 mg/kg depend-ing upon the derivative and the therapeutic schedule which is adopted. Total weekly doses will vary generally between 0.1 and about 35 mg/kg.
The compositiorls in accordance with the inven-tion are prepared as unitary doses of 2 to 900 mg.
The compounds of the invention may be used alone or in combination with other carcinosta-tic agents including, for exampler -the alkylating agents, the an-ti-metabolites such as methotrexate, 5-fluoro-uracil, 6-mercap-topurine, 6-thio-guanine, -the cystosine arabinosides, and antibiotics such as actinomycine D, daunorubicine, adriamycine and cis-diamo-dichloro-platine. In particular, the new vinblastine derivatives may be used in combination.
The following specific examples illustrate in a non-limitative way the process for obtaining the compounds of the invention.
,4Z~
Example_ 1. Prepara-tion of 0--4-deacetyl N-deformyl-vincristine mono-hydrazide A.
985 mg of vincris-tine base are dissolved in a mix-ture of 10 ml anhydrous hydrazine and 10 ml methanol. The reaction mixture is mainta,ined a-t 60C, while stirring, for 22 hours.
' The reaction mixture is -then treated wi-th water and -there-af-ter with wa-ter sa-turated by NaCl. The aqeuous solution is extracted 8 times with 15 ml dichloromethane. The collec-ted organic phases are treated with 20 ml water, -thereafter 25 ml water saturated by NaCl and finally dried on Na2SO4.
After filtration and vacuum concen-tration, 900 mg of 0-4-deacetyl vincristine monohydrazide (purity over 90%) have been obtained.
NMR spectrum (CDC 13,360 MHz) 9.76 (s,lH), 8.37 (s,lH), 8.06 (s,lH), 7.53 (d,lH), 7.24 and 7.07 (m,3H), 6.82 (s,lH), 6.22 (s,l), 5.87 (dd,lh), 5.67 (d,lH), 5.43 ~s,lH), 4.03 (s,lH), 3.94 (s,3H), 3.86 (s,lH), 3.73 (s,3H), 3.52 (s,3H), 2.82 (s,2H), 2.51 (s,lH), 0.93 (2t, 2 x 3H).
Mass Spectrum (elec-tronic ionisa-tion impact) 154, 294, 355, 413, 524, 555, 695, 710, 723, 736, 739, 755 (M 1) - 768, 769, 782 calculated for C42H54N6O7 754 2. Prepara-tion o~ acid azide of modified vincristine A solu-tion of 850 mg of vincristine monohydrazide A in 20 ml methanol is trea-ted with 63 ml HCl 1 N and 175.5 mg sodium nitrite for 15 lminutes at 0C.
;
I'he solution maintained at 0C is then neu-tralised by NaHCO3 at 0C and thereaEter extracted 3 times with dichloromethane.
The collected organic phases are dryed on Na2SO4, filtrated and vacuum concentrated without heating until a solution of about 10 ml is ob-tained.
3. Reaction with ethyl -tryptophana-te.
300 mg of ethyl -tryptophanate are then added while stirring at about 4C and the reaction is checked by tlc. After 10 days, the reaction product is purified by chromatographic separation (40 g silicagel 60, elution solvent ether /NH3 saturated methanol 96:4 volume/volume)~ Fractions of 15 ml are collected which are checked by -tlc. The non-reacted amino-acid is first collec-ted (fraction 30 to 40). The elu-tion agent is changed (ether/NH3 sa-turated methanol 86:14) and -the reacted deriva-tives are collected as -three frac-tions:
fractions 44 and 45:30 m~3; fractions 46-53:257 mg; fraction 54-60:115 mg).
The second part was ethyl N-(0-4-deace-tyl deformyl vincris-tine-23-oyl) tryptophanate Ia which was homogeneous by tlc.
Mass spectru : 984, 970, 956 (M -~ 1) 913, 912, 899 (DCJ
isobu-tane), calculated for C55H66N609 : 955,181.
~V spectrum (CH OH) :
max: 282 (4.20) - 290 (4.18) - 322 (3.96) min: 254 - 287 - 305 IR spectrum: (cm :
_MR Spec-trum (CDC13) 360 MH (ppm) 8.50 (s,lH ),8.03 (s,2H), 7.77 (d,lH), 7.60 (d,lH), 7.57 (d, lH), 6.95 (s,lH), 6.35 (s,lH), 5.89 (d.d.,lH), 5.78 (d,lH), 4.75 (q,lH), 3.88 (s,3H), 3.67 (s,3H), 1.23 (t,3H), 0.98 (t,3il), 0.89 (-t,3H) Ethyl N-(0,4-deacetyl-vincristine-23-oyl)-tryp-tophanate IB.
, The deformylated derivative I a (257 mg) is re-formylated by applying the method described in Belgian Pa-tent 811,110.
The raw product -thus obtained is purified by chromatographic separation using a 20 cm column (silicagel, 30 g) and succe-sively 3 elu-tion agen-ts: ether/NH3 satura-ted MeOH 92:8;
88:12, 86:14. The effluent is collected as 15 ml fractions.
The fractions 44-54 (34.1 mg) contained the N-reformylated derivative Ib.
Mass spectrum (isobu-tane DCI) : 1011,997, 983 (M + 1), 970, 982, 996, 998, 999, 1012 calculated for C56H66N6010 = 983,192 UV spectrum (CH30H) max: 270 (4.18) - 290 (4.11) plate 280 (4.12) shouldering (4.03) IR _pec-trum (KBr) bands at 3400 - 3040 - 2960 - 2920 - 2880 - 2850 - 1735 -1725 - 1680 - 1670 1665 - 1610 - 1490 - 1450 - 1225 - 7i~5.
Example 2 Ethyl N-(0-4 deacetyl-deoxy-4'-vinblas-tine-23-oyl-B) -tryphtophante Ic.
~9~4~Z
The 0-4-deacetyl-deoxy-vinblastine hydrazide is prepared by the me-thod described in U.S. Pa-tent 4,203,898 (assigned to Ely Lilly Co., column 24, line 51 and following).
500 mg of the hydrazide in 12 ml methanol are -thereafter treated wi-th 37 ml HC1 lN and 104 mg NaNO2 for 15 minutes at 0C
The solu-tion is -thereafter neutralized by NaHCO3 and extrac-ted six times with 15 ml dichloromethane. The organic phases are dryed on Na2SO4 and concentrated until a volume of about 10 ml is obtained.
The azide in solution is then brought into con-tact with ethyl tryptophanate (300 mg) at 4C while s-tirring. The reaction is checked by tlc (ether, MeOH). Af-ter 6 days, the reaction is comple-ted and the reaction produc-t is purified by column chroma-tography on silicagel (18 cm, 30 g), elution by 15 ml Eractions.
The following elution agents have been successively used:
E-ther 96 NH3 sat, MeOH 4 frac-tions 1-49 92 8 frac-tions 50-57 86 14 fractions 71-90 The fractions 50-57 contain the wanted reac-tion product Ic (108 mg).
The sulphate is prepared by e-ther precipi-tation (2% H2SO
ethanol).
W spectrum of Sulphate (CH30H) max: 224 (4.63) - 272 (4.32) min: 247 (4.03) , 34~2 shouldering: 286 (4,22) - 312 (3,72) IR spec-trum (KBr) of Sulphate 3420 - 3060 - 2960 - 2930 - 2880 - 2600 (weak) - 1725 -1060 - 1015 - 920 - 745 cm 1, NMR spectrum (CHCl ) MHz of the base in ppm.
9.46 s lH 7.53 d lH 4.94 q lH
8.34 s lH 7.30 d lH 4.18 d lH
7.92 s lH 6.51 s lH 4.05 q 2H
7.67 d 1ll 6.04 s lH 3.77 s 3H
7.59 d lH m. about s. centered on 5.81 - 3.58 s RH
3.47 s lH
2.82 s 2H
2.77 s 3H
2.59 s lH
1.14 t 3H
0.95 t 3H
0.87 t 3H
Mass spectrum of the base DCI isobutane M + 1 at 954 M + 14 + 1 at 968 M + 28 + 1 at 982 ions at 153 - 155 - 157 - 171 - 172 - 185 186 - 199 -calculated for C56 H68 N6 8 Example 3 Etilyl N-(O-4-~eace-tyl-4'-deoxy-vinblas-tin-23-oyl-B so leucinate.
A solution of O-4~deacetyl-4'-deoxyvinblastine hydrazide (0.6, 0.79 mM - see U.S. patent 4,203,898 column 24, line 51) in a mi~-ture of 15 ml of CH30H and 45 ml HCl l N
, was coo]ed to -7~C. NaNO2 was added to the solution (0.15 g) and stirring was continued for 13 minutes a-t -7C. After addi-tion of an aqueous saturated solution of NaHCO3 -till a pH-value of 9 is obtained, -the resulting solu-tion is ex-tracted four times with CH2C12. The combined organic phases were washed wi-th sat.aqueous NaCl solution and dried over ~5 MgSo4.
The solution was concentrated to a volume of 15 ml and ethyl isoleucinate (1.25 mM) was added. The reaction mixture was allowed -to stand 4 days in a refrigerator. The solvent was then evaporated under reduced pressure. The residue was purified by column chromatography (SiO2, 60 g) and eluted with ether/CFl3OH/NH3 (96:4)Ø25 g of amorphous produc-t (yield 35~) have been thus obtained.
Mass spectrum: (Molec.Ionisation isobutane) 880 (M ), 894, 836, 849, 832, 445; 391, 355 cm Melting poin-t 180-190 C
~D = 104 (c=0.154) IR (CHC13) 3470, 2970, 1728, 1655, 1612, 1502 cm UV (CH30H) 289, 268, 216 nm NMR ~CDC13) 7.9 (NH), 6.54 and 6.06 Hg, H12, 5.81 (H14-H15) 4.06 (CHCO2(NH)) 3.76 CO2CH3, 3.58 CH3-O, 3.33 and 2.85 (H3A and H3B), 2.73 (N-CH3), 0.69 (H-15')-!~
~9~22 Acute toxicity-Determination of LDSo The acute -toxicity of the compound of Example 2:
ethyl N-(O-4-deace-tyl-deoxy-4'-vinblastine-23-oyl-B) try-phtophanate has been determined on female NMRI mice. In-creasing doeses of -the drug have been inoculated intravenously in a single injection. The mortality percentage is deter--mined as a functi,on of -the time. The LD50 values for -the ' above compound (deoxy VTrpE) and a reference product: deoxy-vinblastine (deoxy VBL) are indicated hereafter Drug LD50 Deoxy VBL 20 Deoxy VTrpE 91.5 (i.e. Compound The compound deoxy VTrpE is thus less toxic -than the reference compound.
Antitumoral activity The chemiotherapuetic activity of the sulfa-te salt of deoxy VTrpE has been studied on the lymphoblas-tic leu-kemia L 1210 and P 388.
emale DBA2 mice have been inoculated by 104 leu-kemic cells. The produc-t is administered by intravenous route, the day after the graft at a 50mg/kg dose. The mor-tality of the animals is observed as a function of time.
Table I hereafter indica-tes the results obtained wi-th com-pound of Example 2 (Ic). At the tested dose, -the compound is more active than vinblastine. This superiority has been confirmed compared to deoxy-vinblastine.
Female DBA2 mice are in-travenously inocu:lated by 10 leukemic cells. The pro~uc-ts are i.v. adminis-tered, -the day after the graf-t. I'he mortality of -the animals is observed as a func-tion of -time. Resul-ts appear in Table I.
Deoxy VTrpE is clearly more active than the reference com-pound and indica-tes a higher percen-tage of surviving subjects , at long term.
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Claims (22)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A process for preparing compounds of the general formula:
(II) wherein R is an ester of an amino-acid which is attached to the vinblastin-23-oyl moiety by an amide bond, the ester group, which may be straight or branched, containing 2-9 car-bon atoms and R1 is a hydrogen atom or a hydroxy group, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R5 is a hydrogen atom or a methyl group or a formyl group, with the provision that compounds where, simultaneously, R5 is methyl and R1 is a .beta.-hydroxy group are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids comprising reacting a vinblastine derivative of formula II wherein R1, R2 and R5 are as above and R is a methoxy group with a hydrazine, reacting the modified 3-carboxhydrazine vinvlastine so obtained with a nitrosation reagent in an acidic medium, and reacting the corresponding carboxazide so obtained with one to five equivalents of an amino-acid ester containing 2 to 9 carbons in the ester group, and when required, formulating the N-desmethylvinblastine derivative so obtained.
(II) wherein R is an ester of an amino-acid which is attached to the vinblastin-23-oyl moiety by an amide bond, the ester group, which may be straight or branched, containing 2-9 car-bon atoms and R1 is a hydrogen atom or a hydroxy group, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R5 is a hydrogen atom or a methyl group or a formyl group, with the provision that compounds where, simultaneously, R5 is methyl and R1 is a .beta.-hydroxy group are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids comprising reacting a vinblastine derivative of formula II wherein R1, R2 and R5 are as above and R is a methoxy group with a hydrazine, reacting the modified 3-carboxhydrazine vinvlastine so obtained with a nitrosation reagent in an acidic medium, and reacting the corresponding carboxazide so obtained with one to five equivalents of an amino-acid ester containing 2 to 9 carbons in the ester group, and when required, formulating the N-desmethylvinblastine derivative so obtained.
2. A process according to claim 1 for preparing compounds of the general formula:
(III) wherein R1 is a hydroxy group or a hydrogen atom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched C1-C8 alkyl, hydroxy-C1-C8-alkyl, amino-hydroxy-C1-C8-alkyl, carboxy-C1-C8-alkyl, amido-C1-C8-alkyl, guanadino-C1-C8-alkyl, amino-C1-C8-alkyl, thiol-C1-C8-alkyl, cysteinyl-methyl, methylthio-ethyl, benzyl, hydroxy-benzyl, or a group:
(IV) or (V) or R3 together with the carbon to which it is attached and the nitrogen of the amido group, forms an azole or an hydroxy-azole ring; R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched C1-C8-alkyl, or a benzyl group, with the provision that compounds where, simultaneously R1 is .beta.-OH and R5 is -CH3 are excluded; or their pharmaceutically acceptable acid addition salts with mineral or organic acids, comprising reacting a vinblastine derivative of the formula II given in claim 1, wherein R1, R2 and R5 are as above, R is a methoxy group with a hydrazine, reacting the modified 3-carboxyhydrazide vinblastine so obtained with a nitrosation reagent in an acidic medium and reacting the corresponding 3-carboxazide so obtained with one to five equivalents of an amino-acid ester of the formuls NH2CH(R3) COOR4 where R3 and R4 are as above, and when required, formylating the N-desmethylvinblastine derivative so obtained.
(III) wherein R1 is a hydroxy group or a hydrogen atom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched C1-C8 alkyl, hydroxy-C1-C8-alkyl, amino-hydroxy-C1-C8-alkyl, carboxy-C1-C8-alkyl, amido-C1-C8-alkyl, guanadino-C1-C8-alkyl, amino-C1-C8-alkyl, thiol-C1-C8-alkyl, cysteinyl-methyl, methylthio-ethyl, benzyl, hydroxy-benzyl, or a group:
(IV) or (V) or R3 together with the carbon to which it is attached and the nitrogen of the amido group, forms an azole or an hydroxy-azole ring; R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched C1-C8-alkyl, or a benzyl group, with the provision that compounds where, simultaneously R1 is .beta.-OH and R5 is -CH3 are excluded; or their pharmaceutically acceptable acid addition salts with mineral or organic acids, comprising reacting a vinblastine derivative of the formula II given in claim 1, wherein R1, R2 and R5 are as above, R is a methoxy group with a hydrazine, reacting the modified 3-carboxyhydrazide vinblastine so obtained with a nitrosation reagent in an acidic medium and reacting the corresponding 3-carboxazide so obtained with one to five equivalents of an amino-acid ester of the formuls NH2CH(R3) COOR4 where R3 and R4 are as above, and when required, formylating the N-desmethylvinblastine derivative so obtained.
3. The process of claim 2, wherein the nitrosa-tion is effected with sodium nitrite in the presence of methanolic HC1.
4. A process according to claim 1, in which the azide of O-4-deacetyl-deoxy-4'-vinblastin-23-oic acid is reacted with one to five equivalents of ethyl-L-tryptophanate.
5. A process according to claim 1, in which the azide of O-4-deacetyl-deoxy-4'-vinblastin-23-oic acid is reacted with one to five equivalents of methyl-L-tryptophanate.
6. A process according to claim 1, in which the azide of O-4-deacetyl-vincristin-23-oic acid is reacted with one to five equivalents of ethyl-L-tryptophanate.
7. A process according to claim 1, in which the azide of O-4-deacetyl-vincristin-23-oic acid is reacted with one to five equivalents of methyl-L-tryptophanate.
8. A process according to claim 1, in which the azide of O-4-deacetyl-deformyl-vincristin-23-oic acid is reacted with one to five equivalents of ethyl-L-tryptophanate.
9. A process according to claim 1, in which the azide of O-4-deacetyl-deformyl-vincristin-23-oic acid is reacted with one to five equivalents of methyl-L-tryptophanate.
10. A process according to claim 2, wherein a 1:1 addition salt with sulfuric acid is obtained.
11. Vinblastine derivatives of the general formula (II) wherein R is an ester of an amino -acid attached to the vin-blastin-23-oyl moiety by an amide bond, the ester group, which may be straight or branched, containing 2-9 carbon atoms and R1 is a hydrogen atom or a hydroxy group, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R5 is a hydrogen atom or a methyl group or a formyl group, with the provision that compounds where, simultaneously, R5 is methyl and R1 is a .beta.-hydroxy group are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids when-ever prepared or produced by the process claimed in claim 1 or an obvious chemical equivalent thereof.
12. Vinblastine derivatives of the general formula:
(III) wherein R1 is a hydroxy group or a hydrogen atom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched C1-C8 alkyl, hydroxy-C1-C8-alkyl, amino-hydroxy-C1-C8-alkyl, carboxy-C1-C8-alkyl, amido-C1-C8-alkyl, guanadino-C1-C3-alkyl, amino-C1-C8-alkyl, thiol-C1-C8-alkyl, cysteinyl-methyl, methylthio-ethyl, benzyl, hydroxy-benzyl, or a group:
(IV) or (V) or R3 together with the carbon to which it is attached and the nitrogen of the amido-group, forms an azole or an hydroxy-azole ring, R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched C1 -C8-alkyl, or a benzyl group, with the provision that compounds where, simul-taneously R1 is .beta.-OH and R5 is -CH3 are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids, whenever prepared or produced by the process claimed in claim 2 or an obvious chemical equivalent thereof.
(III) wherein R1 is a hydroxy group or a hydrogen atom, R2 is a hydrogen atom, a formyl group or a C2-C9 acyl group, R3, considered each time independantly, is a hydrogen atom, straight or branched C1-C8 alkyl, hydroxy-C1-C8-alkyl, amino-hydroxy-C1-C8-alkyl, carboxy-C1-C8-alkyl, amido-C1-C8-alkyl, guanadino-C1-C3-alkyl, amino-C1-C8-alkyl, thiol-C1-C8-alkyl, cysteinyl-methyl, methylthio-ethyl, benzyl, hydroxy-benzyl, or a group:
(IV) or (V) or R3 together with the carbon to which it is attached and the nitrogen of the amido-group, forms an azole or an hydroxy-azole ring, R5 is a hydrogen atom or a methyl group or a formyl group and R4 is a straight or branched C1 -C8-alkyl, or a benzyl group, with the provision that compounds where, simul-taneously R1 is .beta.-OH and R5 is -CH3 are excluded; or their pharmaceutically acceptable addition salts with mineral or organic acids, whenever prepared or produced by the process claimed in claim 2 or an obvious chemical equivalent thereof.
13. Ethyl-N-(O-4-deacetyl-deoxyvinblastin-23-.beta.-oyl)-L-tryptophanate or an addition salt thereof with pharmaceu-tically acceptable acids whenever prepred or produced by the process claimed in claim 4 or an obvious chemical equivalent thereof.
14. Methyl N-(O-4-deacetyl-4-deoxyvinblastin-23-oyl-B)-L-tyrptophanate or an addition salt thereof with phar-maceutically acceptable acids whenever prepared or produced by the process claimed in claim 5 or an obvious chemical equivalent thereof.
15. Methyl N-(O-4-deacetyl-vincristin-23-oyl)-L-tryptophanate or addition salt thereof with pharamaceutically acceptable acids whenever prepared or produced by the process claimed in claim 7 or an obvious chemical equivalent thereof.
16. Ethyl N-(O-4-deacetyl-vincristin-23-oyl)-L-tryptophanate or an addition salt thereof with pharmaceu-tically acceptable acids whenever prepared or produced by the process claimed in claim 6 or an obvious chemical equivalent thereof.
25.
25.
17. Methyl N-(O-4-deacetyl-deformyl-vincristin-23-oyl)-L- tryptophanate or an addition salt thereof with pharma-ceutically acceptable acids whenever prepared or produced by the process claimed in claim 9 or an obvious chemical equivalent thereof.
18. Ethyl N-(O-4-deacetyl-deformyl-vincrinstin-23-oyl)-L-tryptophanate or an addition salt thereof with pharma-ceutically acceptable acids whenever prepared or produced by the process claimed in claim 8 or an obvious chemical equi-valent thereof.
19. An acid addition salt of a compound of formula III given in claim 2 with sulphuric acid whenever prepared or produced by the process claimed in claim 10 or an obvious chemical equivalent thereof.
20. A process according to claim 2, in which the amino acid ester is an ester of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, gluta-mic acid, aspargine, glutamine, arginine, lysine, cysteine, cystine, methionine, phenylalamine, tyrosine, tryptophan , proline, histidine, hydroxy-lysine or hydroxy-proline.
21. A process according to claim 2, in which the amino acid ester is an ester of L or D tryptophan, leucine, isoleucine, valine, alanine or phenyl-alanine.
22. A process according to claim 2, in which the amino acid ester is an ester of L-tryptophan , L-isolelucine and L-alanine.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU83.822 | 1981-12-08 | ||
| LU83822A LU83822A1 (en) | 1981-12-08 | 1981-12-08 | N- (VINBLASTINOYL-23) DERIVATIVES OF AMINO ACIDS, THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATION |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1198422A true CA1198422A (en) | 1985-12-24 |
Family
ID=19729780
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000416996A Expired CA1198422A (en) | 1981-12-08 | 1982-12-03 | N-(vinblastinoyl-23) derivatives of amino acids, their preparation and therapeutical application |
Country Status (24)
| Country | Link |
|---|---|
| JP (1) | JPS58116491A (en) |
| AT (1) | AT390958B (en) |
| AU (1) | AU564951B2 (en) |
| BE (1) | BE895262A (en) |
| CA (1) | CA1198422A (en) |
| CH (1) | CH655727A5 (en) |
| DD (1) | DD205906A5 (en) |
| DE (1) | DE3245269A1 (en) |
| DK (1) | DK161833C (en) |
| ES (1) | ES8401768A1 (en) |
| FR (1) | FR2517680B1 (en) |
| GB (1) | GB2111055B (en) |
| GR (1) | GR77280B (en) |
| HU (1) | HU187803B (en) |
| IE (1) | IE55283B1 (en) |
| IL (1) | IL67366A (en) |
| IT (1) | IT1157126B (en) |
| LU (1) | LU83822A1 (en) |
| NL (1) | NL8204711A (en) |
| NZ (1) | NZ202708A (en) |
| OA (1) | OA07270A (en) |
| PT (1) | PT75949B (en) |
| SE (1) | SE452622B (en) |
| ZA (1) | ZA828823B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0124502B1 (en) * | 1983-04-29 | 1991-06-12 | OMNICHEM Société anonyme | Conjugates of vinblastine and its derivatives, process for their preparation and pharmaceutical compositions containing these conjugates |
| US4667030A (en) * | 1985-06-17 | 1987-05-19 | Eli Lilly And Company | Hydrazide succinimide derivatives of antineoplastic indole-dihydroindole alkaloids |
| US4675400A (en) * | 1985-06-17 | 1987-06-23 | Eli Lilly And Company | Bifunctional derivatives of 4-desacetyl indole-dihydroindole alkaloids |
| JPH075608B2 (en) * | 1986-01-13 | 1995-01-25 | イ−ル−セルタ−ク ソシエテ アノニム | Vinblastine derivative |
| US4906086A (en) * | 1987-06-19 | 1990-03-06 | Honda Giken Kogyo Kabushiki Kaisha | Rearview mirror device for motorcycles |
| FR2626882B1 (en) * | 1988-02-08 | 1991-11-08 | Ire Celltarg Sa | VINCA DERIVATIVE CONJUGATES COMPRISING A DETERGENT CHAIN IN POSITION C-3 |
| JP6250710B2 (en) * | 2013-04-19 | 2017-12-20 | 曁南大学 | Preparation method of vinblastine derivative |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AR204004A1 (en) | 1973-04-02 | 1975-11-12 | Lilly Co Eli | PROCEDURES FOR PREPARING VINBLASTIN LEUROSIDINE AND LEUROCRISTINE DERIVATIVES |
| US3848234A (en) * | 1973-04-04 | 1974-11-12 | Sperry Rand Corp | Multi-processor system with multiple cache memories |
| IL48685A (en) * | 1975-01-09 | 1980-03-31 | Lilly Co Eli | Amides of vincadioline and vinblastine |
| OA06421A (en) * | 1980-06-10 | 1981-09-30 | Omnium Chimique Sa | Process for the preparation of N- (vinblastinoyl-23) derivatives of amino acids and peptides. |
-
1981
- 1981-12-08 LU LU83822A patent/LU83822A1/en unknown
-
1982
- 1982-11-24 AU AU90847/82A patent/AU564951B2/en not_active Ceased
- 1982-11-30 IL IL67366A patent/IL67366A/en unknown
- 1982-12-01 ZA ZA828823A patent/ZA828823B/en unknown
- 1982-12-01 CH CH6980/82A patent/CH655727A5/en not_active IP Right Cessation
- 1982-12-03 NZ NZ202708A patent/NZ202708A/en unknown
- 1982-12-03 CA CA000416996A patent/CA1198422A/en not_active Expired
- 1982-12-06 NL NL8204711A patent/NL8204711A/en not_active Application Discontinuation
- 1982-12-06 GR GR69997A patent/GR77280B/el unknown
- 1982-12-06 ES ES517942A patent/ES8401768A1/en not_active Expired
- 1982-12-07 DK DK541882A patent/DK161833C/en not_active IP Right Cessation
- 1982-12-07 BE BE0/209661A patent/BE895262A/en not_active IP Right Cessation
- 1982-12-07 JP JP57214573A patent/JPS58116491A/en active Granted
- 1982-12-07 HU HU823919A patent/HU187803B/en unknown
- 1982-12-07 AT AT0444982A patent/AT390958B/en not_active IP Right Cessation
- 1982-12-07 PT PT75949A patent/PT75949B/en not_active IP Right Cessation
- 1982-12-07 DE DE19823245269 patent/DE3245269A1/en not_active Withdrawn
- 1982-12-07 IE IE2906/82A patent/IE55283B1/en not_active IP Right Cessation
- 1982-12-07 FR FR828220476A patent/FR2517680B1/en not_active Expired - Lifetime
- 1982-12-07 IT IT68431/82A patent/IT1157126B/en active
- 1982-12-07 SE SE8206981A patent/SE452622B/en not_active IP Right Cessation
- 1982-12-08 GB GB08234965A patent/GB2111055B/en not_active Expired
- 1982-12-08 OA OA57864A patent/OA07270A/en unknown
- 1982-12-08 DD DD82245703A patent/DD205906A5/en not_active IP Right Cessation
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