DE10008089A1 - Production of tubulysin compounds comprises multi-stage process including condensation of N-methylpipecolinoyl-isoleucine with substituted thiazole-4-carboxylic acid derivative - Google Patents

Production of tubulysin compounds comprises multi-stage process including condensation of N-methylpipecolinoyl-isoleucine with substituted thiazole-4-carboxylic acid derivative

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DE10008089A1
DE10008089A1 DE10008089A DE10008089A DE10008089A1 DE 10008089 A1 DE10008089 A1 DE 10008089A1 DE 10008089 A DE10008089 A DE 10008089A DE 10008089 A DE10008089 A DE 10008089A DE 10008089 A1 DE10008089 A1 DE 10008089A1
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Gerhard Hoefle
Thomas Leibold
Heinrich Steinmetz
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HELMHOLTZ-ZENTRUM FUER INFEKTIONSFORSCHUNG GMBH, DE
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    • C07D207/18—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member
    • C07D207/22—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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Abstract

Tubulysin compounds are produced by a multi-stage process using known and new starting materials and intermediates. Production of tubulysin compounds (I) comprises: (A) reaction of a N-methyl-pipecolinic acid (II) with isoleucine benzyl ester (III) to give N-methylpipecolinoyl-isoleucine (IV); (B) reaction of a substituted thiazole-4-carboxylic acid (V) with a substituted pentanoic acid derivative (VI) to give a 2-(substituted pentanoyl)-thiazole-4-carboxylic acid (VII); and (C) reaction of compound (IV) with compound (VII) to give a tubulysin precursor (VIII) which is converted into a tubulysin compound (I) by: (i) conversion of the 1'-keto group into acetyl; (ii) separation of diastereomers; and (iii) conversion of TMSE (undefined) into COOH. R<1> = H; alkyl; aryl; or heteroaryl; R<2> = phenyl or 4-OH-phenyl. Independent claims are also included for: (1) compounds of formulae (V), (VI), (IX), (X) (as the L-isomer or racemate) and (XI) and as well as their preparation; and (2) the preparation of compounds (IIA).

Description

Erfindungsgemäß wird die absolute Konfiguration von Tubulysin, ein Verfahren zur Herstellung von Tubulysinen und die dabei eingesetzten Zwischenprodukte offenbart.According to the invention, the absolute configuration of tubulysin, a process for the production of tubulysins and the intermediates used disclosed.

Vor kurzem haben wir die Tubulysine (1) als eine neue, auf das Tubulin-Skelett wirkende, Substanzfamilie aus Myxobakterien in Irsee vorgestellt [1]. Im Gegensatz zu den Epothilonen zeigen diese eine mikrotubuli-abbauende Wirkung sowie die vermehrte Ausbildung von Zentrosomen [1] (s. Abb. 1). Mit einer Cytotoxi­ zität von IC50 = 10-500 pg sind die Tubulysine als potentiel­ le Cytostatika von besonderem Interesse.We recently introduced tubulysins (1) as a new family of substances from myxobacteria in Irsee that acts on the tubulin skeleton [1]. In contrast to the epothilones, these show a microtubule-degrading effect and the increased formation of centrosomes [1] (see Fig. 1). With a cytotoxicity of IC 50 = 10-500 pg, the tubulysins are of particular interest as potential cytostatics.

Die Aufklärung der Konstitution erfolgte mit spektroskopischen Methoden, insbesondere 2D-NMR-Techniken, und durch Einbau von 13C-markiertem Acetat und Methionin. Aufgrund der außergewöhn­ lichen Aktivität und einer interessanten Ähnlichkeit zu dem analog wirkenden Dolastatin 10[2a] (s. Abb. 2) wurde ein Pro­ gramm zur Aufklärung der absoluten Konfiguration sowie an­ schließender Totalsynthese des Tetrapeptids in Angriff genom­ men. The constitution was elucidated using spectroscopic methods, in particular 2D NMR techniques, and by incorporating 13 C-labeled acetate and methionine. Due to the extraordinary activity and an interesting similarity to the analogous dolastatin 10 [2a] (see Fig. 2), a program was initiated to elucidate the absolute configuration and subsequent total synthesis of the tetrapeptide.

Die Konfigurationszuordnung erfolgte durch Vergleich der durch Totalhydrolyse erhaltenen Aminosäuren mit den entsprechenden Synthese-Intermediaten mittels GC- und HPLC-Analytik. Die Tu­ bulysine enthalten nur eine proteinogene (natürliche) Aminosäu­ re, Isoleucin. Für die Synthese der drei weiteren Bestandteile wurde erfindungsgemäß stets ein enantio- bzw. diastereoselekti­ ver Syntheseweg entwickelt.The configuration assignment was made by comparing the Total hydrolysis obtained amino acids with the corresponding Synthesis intermediates using GC and HPLC analysis. The Tu bulysine contain only one proteinogenic (natural) amino acid right, isoleucine. For the synthesis of the three other components was always an enantio- or diastereoselecti according to the invention ver developed synthetic route.

Es zeigen:Show it:

Fig. 1: Vermehrte Ausbildung von Zentrosomen und Abbau der Mi­ krotubuli bei Tubulysin D Zugabe (0,5 ng/ml, PtK2 Zellen der Beutelratte); Fig. 1: Increased formation of centrosomes and degradation of the Mi krotubuli with Tubulysin D addition (0.5 ng / ml, PtK 2 cells of the bag rat);

Fig. 2: Tubulysine A-F, Biogenesestudie und Vergleich mit Do­ lastatin 10 bzw. Lu 103793 [2b]; Fig. 2: Tubulysine AF, biogenesis study and comparison with Do lastatin 10 and Lu 103793 [2b];

Fig. 3a: 'H-NMR-Spektrum von Tubulysin A (MSO, 400 MHz); Fig. 3a: 'H-NMR spectrum of tubulysin A (MSO, 400 MHz);

Fig. 3b: 13C-NMR-Spektrum von Tubulysin A (DMSO, 400 MHz); FIG. 3b: 13 C-NMR spectrum of Tubulysin A (DMSO, 400 MHz);

Fig. 4: GC-Spektren von Baustein I (FS-Hydrodex β-3P, 25 m, 120°C); Fig. 4: GC spectra of block I (FS-Hydrodex-β 3P, 25 m, 120 ° C);

Fig. 5: GC-Spektren von Baustein II (Permabond L-Chirasil-Val, 25 m, 80°C); Fig. 5: GC spectra of Block II (Permabond L-Chirasil-Val, 25 m, 80 ° C);

Fig. 6: GC-Spektren von Baustein IV (FS-Hydrodex β-3P, 25 m, 165°C); Fig. 6: GC spectra of block IV (FS-Hydrodex-β 3P, 25 m, 165 ° C);

Fig. 7: Relative Konfiguration von 16 (δ in ppm) (W. A. König et al. Liebigs Ann. Chem., 1987, 803-807); Fig. 7: Relative configuration of 16 (δ in ppm) (WA Koenig et al. Liebigs Ann. Chem., 1987, 803-807);

Fig. 8: GC-Spektren von Baustein III (Permabond L-Chirasil-Val, 25 m, 80°C); Fig. 8: GC spectra of Component III (Permabond L-Chirasil-Val, 25 m, 80 ° C);

Fig. 9: Absolute Konfiguration von Tubulysin 1; Fig. 9: Absolute configuration of tubulysin 1;

Synthese der AninosäurebausteineSynthesis of the amino acid building blocks Baustein I: N-Methyl-pipecolinsäure (Mep)Module I: N-methyl-pipecolic acid (Mep)

Enantiomerenreine N-Methyl-pipecolinsäure 3 wurde durch reduk­ tive Aminierung von L-Pipecolinsäure 2 dargestellt (s. Schema 1). Racemische N-Methyl-pipecolinsäure 3* wurde durch Versei­ fung von käuflichem Ethylester-Racemat hergestellt.Enantiomerically pure N-methyl-pipecolic acid 3 was by reduk active amination of L-pipecolic acid 2 (see scheme 1). Racemic N-methyl-pipecolic acid 3 * was obtained by Versei tion of commercially available ethyl ester racemate.

Schema 1 Synthese von L-N-Methyl-pipecolinsäure Scheme 1 Synthesis of LN-methyl-pipecolic acid

Baustein II: Isoleucin (Ile, allo-Ile)Module II: isoleucine (Ile, allo-Ile)

Alle GC-Derivate wurden direkt aus den käuflichen Isoleucin- bzw. allo-Isoleucin-Enantiomeren dargestellt.All GC derivatives were obtained directly from the commercially available isoleucine or allo-isoleucine enantiomers.

Baustein III: 2-(3-Amino-1-hydroxy-4-methyl-pentyl)-thiazol-4- carbonsäure (Tubuvalin, Tuv)Module III: 2- (3-amino-1-hydroxy-4-methylpentyl) thiazole-4- carboxylic acid (tubuvaline, Tuv)

Ausgehend von N-Cbz-L-Valin 4 wurde der Aminoalkohol 5 darge­ stellt, der nach Swern-Oxidation mit dem Thiazol-Wittig-Ylid 6 zum Enolether 7 umgesetzt wurde. Starting from N-Cbz-L-valine 4, the amino alcohol 5 was shown after Swern oxidation with the thiazole-Wittig ylide 6 was converted to enol ether 7.  

Saure Hydrolyse des Enolethers lieferte Keton 8, welches durch Reduktion mit Natriumborhydrid in die diastereomeren, geschütz­ ten Tuv-Analoga 9a/b überführt wurde. Entsprechend wurde auch das Racemat 8* erhalten (s. Schema 2)Acid hydrolysis of the enol ether gave ketone 8, which by Reduction with sodium borohydride in the diastereomeric, protected ten Tuv analogues 9a / b was transferred. Correspondingly, too receive the racemate 8 * (see scheme 2)

Schema 2 Darstellung des Bausteins III (9) Scheme 2 Representation of building block III (9)

Baustein IV: 4-Amino-5-phenyl-2-methyl-pentansäure (Tubuphe­ nylalanin, Tup)Component IV: 4-amino-5-phenyl-2-methylpentanoic acid (Tubuphe nylalanine, Tup)

N-BOG-L-Phenylalanin 10 wurde durch Reduktion, Reoxidation mit­ tels Swern-Oxidation und abschließende Wittig-Reaktion um eine C3-Einheit verlängert. Es wurden zwei Produkte erhalten, bei denen es sich um das offenkettige E-Additionsprodukt 13 sowie das Lactam 12 handelte. Chromatographische Trennung und Hydrie­ rung lieferte aus 13 ein 2 : 1 Isomerengemisch 15a/b, während aus 12 nur das BOC-geschützte Lactam 14 mit 2R/4R-Konfiguration entstand. Nach Ringöffnung und Veresterung mit Diazomethan wur­ de hieraus diastereomerenreines 15b erhalten (s. Schema 3).N-BOG-L-phenylalanine 10 was extended by a C 3 unit by reduction, reoxidation by means of Swern oxidation and a final Wittig reaction. Two products were obtained, which were the open-chain E addition product 13 and the lactam 12. Chromatographic separation and hydrogenation yielded a 2: 1 isomer mixture 15a / b from 13, while from 12 only the BOC-protected lactam 14 with 2R / 4R configuration resulted. After ring opening and esterification with diazomethane, diastereomerically pure 15b was obtained (see Scheme 3).

Schema 3 Aufbau von Baustein IV ausgehend von Phenylalanin Scheme 3 Structure of building block IV starting from phenylalanine

Konfigurationsaufklärung durch GC-Vergleich an chiralen PhasenConfiguration clarification by GC comparison on chiral phases Totalhydrolyse und DerivatisierungTotal hydrolysis and derivatization

Tubulysin wurde für 12 h mit 6 N Salzsäure, bzw. für die Bestim­ mung von Baustein IV mit Hydrazin-Hydrat bei 100°C abgebaut. Anschließende Veresterung mit methanolischer bzw. ethanolischer Salzsäure bei 100°C und Acylierung mit Trifluoressigsäureanhy­ drid lieferte die für die Messungen verwendeten Derivate. Die synthetisch aufgebauten Vergleichssubstanzen wurden unter iden­ tischen Bedingungen derivatisiert.Tubulysin was for 12 h with 6 N hydrochloric acid, or for the determ Mining of building block IV with hydrazine hydrate at 100 ° C degraded. Subsequent esterification with methanolic or ethanolic Hydrochloric acid at 100 ° C and acylation with trifluoroacetic acid drid supplied the derivatives used for the measurements. The Synthetic reference substances were identified under iden derivatized conditions.

Da bei saurer Hydrolyse eine teilweise Epimerisierung an C-2 des Bausteins IV aufgetreten war, wurde für die Bestimmung der C-terminalen Aminosäure Hydrazin-Hydrat, ohne beobachtbare Epi­ merisierung, eingesetzt.Since, in the case of acid hydrolysis, partial epimerization at C-2 of module IV had occurred, was used to determine the C-terminal amino acid hydrazine hydrate, with no observable epi merization, used.

Konfigurationsaufklärung von Baustein IIIConfiguration elucidation of component III

Die Zuordnung der absoluten Konfiguration von Baustein III sollte zunächst wie bereits für die anderen Bausteine beschrie­ ben erfolgen. Überraschenderweise wurde nach der Derivatisie­ rung von reinem 9a eine starke Racemisierung des benzylischen Zentrums festgestellt (20-40%). Dies war bei der Derivati­ sierung von Tubulysin ebenfalls zu beobachten (vollständige Ra­ cemisierung), weshalb eine direkte Zuordnung auf diesem Weg nicht möglich war.The assignment of the absolute configuration of module III should first be described as for the other building blocks ben done. Surprisingly, after the derivatization tion of pure 9a, a strong racemization of the benzylic Center determined (20-40%). This was with the derivatives of tubulysin can also be observed (complete Ra cemization), which is why a direct assignment in this way was not possible.

Im Verlauf der Hydrolyse der beiden O-Acylgruppen konnten wir ein Tubulysin-Derivat 16 mit cyclischem N,O-Acetal isolieren, welches zweifelsfrei zumindest die relative Konfigurationszu­ ordnung der beiden Zentren ermöglichte (s. Schema 4). In the course of the hydrolysis of the two O-acyl groups, we were able to isolate a tubulysin derivative 16 with cyclic N, O-acetal, which undoubtedly at least the relative configuration order of the two centers enabled (see Scheme 4).  

Schema 4 Synthese eines Tubulysin-Derivats mit cyclischem Ace­ tal Scheme 4 Synthesis of a tubulysin derivative with cyclic acetal

Durch Vergleich der NMR-Daten von 16 mit Literaturwerten wurde für Baustein III eine relative trans-Konfiguration abgeleitet (s. Abb. 7).By comparing the NMR data of 16 with literature values, a relative trans configuration was derived for module III (see Fig. 7).

Der Versuch, die noch fehlende absolute Konfiguration an C-3' durch Vergleich auf der Keto-Stufe 8 (s. Schema 5) durchzufüh­ ren, scheiterte, da auf verschiedenen chiralen GC- und HPLC- Säulen keine Trennung der Enantiomeren erreicht werden konnte. Ein weiterer Abbau sollte nun durch Bayer-Villiger-Oxidation/ -Hydrolyse und Vergleich der Produkte mit Valinol bzw. Homova­ lin erfolgen. Da die Reaktion selbst mit Pertrifluoressigsäure extrem langsam verlief (< 2% Umsatz in 7d), wurde ein ozonoly­ tischer Abbau untersucht. Umsetzung von 8 mit Trimethylsilyl­ trifludrmethansulfonat lieferte zunächst Silylenolether 17, der nach Ozonolyse und oxidativer Aufarbeitung sowie Veresterung zu N-Cbz-Valinethylester abgebaut wurde (s. Schema 5). Attempting the missing absolute configuration at C-3 ' by comparison at keto level 8 (see Scheme 5) failed because of different chiral GC and HPLC Columns no separation of the enantiomers could be achieved. Further degradation should now take place through Bayer Villiger oxidation / Hydrolysis and comparison of the products with Valinol or Homova lin done. Because the reaction itself with pertrifluoroacetic acid was extremely slow (<2% conversion in 7d), became an ozonoly table degradation examined. Reaction of 8 with trimethylsilyl trifludrmethanesulfonat initially provided Silylenolether 17, the after ozonolysis and oxidative workup as well as esterification N-Cbz-valine ethyl ester was degraded (see Scheme 5).  

Schema 5 Abbauversuche ausgehend von Keton 8 Scheme 5 degradation tests based on ketone 8

Durch Anwendung dieser Abbau-Sequenz auf die aus Tubulysin her­ gestellte Aminosäure 19 konnte nun dem letzten Stereozentrum die L-Konfiguration zugeordnet werden (s. Schema 6 und Abb. 8). By applying this degradation sequence to the amino acid 19 made from tubulysin, the L configuration could now be assigned to the last stereo center (see Scheme 6 and Fig. 8).

Schema 6 Abbau von Tubulysin zu 20 Scheme 6 Degradation of tubulysin to 20

Aus den Ergebnissen der GC-Untersuchungen ergibt sich für Tu­ bulysin folgende absolute Konfiguration (s. Abb. 9):The following absolute configuration results for tu bulysin from the results of the GC investigations (see Fig. 9):

Zunächst sollte eine konvergente Totalsynthese von Tubulysin D unter weitgehender Verwendung der bisher dargestellten Ami­ nosäuren durchgeführt werden (s. Schema 9). N-Methyl­ pipecolinsäure wurde in racemischer Form eingesetzt, eine Tren­ nung erfolgte dann auf der Dipeptid-Stufe 27a/b. Baustein III wurde wie bereits beschrieben synthetisiert, der entscheidende Punkt ist hier der selektive Aufbau des Amidoacetals (s. Schema 7). Es ist vorgesehen, die 2'-Hydroxyfunktion im Baustein erst auf einer möglichst späten Stufe durch Reduktion des Ketons einzuführen.First, a convergent total synthesis of tubulysin D largely using the Ami shown so far no acids are carried out (see Scheme 9). N-methyl Pipecolic acid was used in racemic form, a Tren This was then done at dipeptide level 27a / b. Module III  was synthesized as already described, the decisive one The point here is the selective structure of the amidoacetal (see diagram 7). It is envisaged that the 2'-hydroxy function in the building block first at a late stage by reducing the ketone introduce.

Schema 7 Synthese von Baustein-II 22 Scheme 7 Synthesis of Building Block II 22

Die Synthese von Baustein IV 25 wird durch Methylierung der Aminosäure in 2-Position unter Verwendung eines Evans-Auxiliars erreicht (s. Schema 8).The synthesis of building block IV 25 is carried out by methylation of the Amino acid in the 2-position using an Evans auxiliary reached (see Scheme 8).

Schema 8 Synthese von Baustein-IV 25 Scheme 8 Synthesis of building block IV 25

Schema 9 Kupplungsstrategie Scheme 9 coupling strategy

Abschließende Reduktion und Acetylierung an C-1' von Tuv, Tren­ nung der Diastereomeren und Abspaltung des TMSE-esters liefert 1. Final reduction and acetylation at C-1 'by Tuv, Tren of the diastereomers and cleavage of the TMSE ester 1.  

Reagenzien auf S. 7Reagents on p. 7

DECP/NEt3 DECP / NEt 3

: Phosphonsäurediethylester-cyanid/Triethylamin
PFPOH/DCC: Pentafluorphenol/Dicyclohexylcarbodiimid
PEPOTFA: Pentafluorphenol-Trifluoracetat
Ac2 : Diethyl cyanide / triethylamine
PFPOH / DCC: pentafluorophenol / dicyclohexylcarbodiimide
PEPOTFA: Pentafluorophenol trifluoroacetate
Ac 2

O: Essigsäureanhydrid
TBAF: Tetrabutylammoniumfluorid: Acetic anhydride
TBAF: tetrabutylammonium fluoride

Es ist uns gelungen, die absolute Konfiguration von Tubulysin durch Abbau und Vergleich mit Referenzsubstanzen aufzuklären. Die für die vier Aminosäure-Bausteine entwickelten Synthesen erlauben nun eine konvergente Totalsynthese der durch Isolie­ rung nur in äußerst geringen Mengen zugänglichen natürlichen Tubulysine A-F. Darüber hinaus erlaubt es diese Strategie, durch Variation der vier Bausteine eine Bibliothek von stereoi­ someren und strukturanalogen Tubulysinen herzustellen.We have succeeded in the absolute configuration of tubulysin to be clarified by degradation and comparison with reference substances. The syntheses developed for the four amino acid building blocks now allow a convergent total synthesis by isolation only accessible in extremely small quantities Tubulysins A-F. In addition, this strategy allows by variation of the four building blocks a stereoi library to produce other and structurally analogous tubulysins.

Literaturliterature

1) Sasse et al. (Poster) 10. Irseer Naturstofftage der Dechema e. V., Irsee, 1998.
2) a. G. R. Pelitt et al., J. Nat. Prod. 1981, 44, 482-485; b. M. DeArruda et al., Cancer Res. 1995, 55, 3085-3092.
3) J. Lukas et al. Collect. Czech. Chem. Commun. 1957, 22, 286.
4) A. M. P. Koskinen et al. J. Org. Chem. 1998, 63, 92-98. 1) Sasse et al. (Poster) 10th Irseer Naturstoffage der Dechema e. V., Irsee, 1998.
2) a. GR Pelitt et al., J. Nat. Prod. 1981, 44, 482-485; b. DeArruda, M. et al., Cancer Res. 1995, 55, 3085-3092.
3) J. Lukas et al. Collect. Czech. Chem. Commun. 1957, 22, 286.
4) AMP Koskinen et al. J. Org. Chem. 1998, 63, 92-98.

In der Beschreibung und den Ansprüchen können R, R1 und R2 un­ abhängig voneinander ein H-Atom, eine Alkylgruppe, eine Aryl­ gruppe oder eine Heteroarylgruppe darstellen.In the description and claims, R, R1 and R2 can un depending on each other, an H atom, an alkyl group, an aryl represent a group or a heteroaryl group.

Vorzugsweise ist eine Alkylgrppe eine C1-6-Alkylgruppe, weist eine Arylgruppe bis zu vier gegebenenfalls annelierte Ringe auf und weist eine Heteroarylgruppe bis zu vier gegebenenfalls an­ nelierte Ringe auf, wobei jeder Ring bis zu drei Heteroatome wie z. B. N, O oder S-Atome enthalten kann.Preferably an alkyl group is a C1-6 alkyl group an aryl group up to four optionally fused rings and optionally assigns a heteroaryl group up to four fused rings, each ring having up to three heteroatoms such as B. can contain N, O or S atoms.

Claims (35)

1. Verfahren zur Herstellung einer Verbindung der Formel worin R1 ein H-Atom, eine Alkylgruppe, eine Arylgruppe oder eine Heteroarylgruppe ist und R2 ein H-Atom oder eine OH-Gruppe ist,
dadurch gekennzeichnet, daß
  • a) eine Verbindung der Formel 3*
    mit einer Verbindung der Formel 26
    zu einer Verbindung der Formel 27b
    umgesetzt wird;
  • b) eine Verbindung der Formel 22
    in der R1 ein H-Atom, eine Alkylgruppe, eine Arylgruppe oder eine Heteroarylgruppe ist
    mit einer Verbindung der Formel 25
    zu einer Verbindung der Formel 28
    umgesetzt wird;
  • c) die Verbindung der Formel 27b mit der Verbindung der Formel 28 zu einer Verbindung der Formel 29
    umgesetzt wird;
  • d) die Ketogruppe am 1'-C-Atom der Verbindung der Formel 29 in eine Acetylgruppe umgewandelt wird;
  • e) die Diastereomeren getrennt werden, und
  • f) der TMSE-Ester in eine COOH-Gruppe umgewandelt wird.
1. Process for the preparation of a compound of formula where R 1 is an H atom, an alkyl group, an aryl group or a heteroaryl group and R 2 is an H atom or an OH group,
characterized in that
  • a) a compound of the formula 3 *
    with a compound of formula 26
    to a compound of formula 27b
    is implemented;
  • b) a compound of formula 22
    in which R 1 is an H atom, an alkyl group, an aryl group or a heteroaryl group
    with a compound of formula 25
    to a compound of formula 28
    is implemented;
  • c) the compound of formula 27b with the compound of formula 28 to a compound of formula 29
    is implemented;
  • d) the keto group on the 1'-C atom of the compound of the formula 29 is converted into an acetyl group;
  • e) the diastereomers are separated, and
  • f) the TMSE ester is converted into a COOH group.
2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß in Schritt a) Phosphonsäurediethylester-cyanid/Triethylamin eingesetzt wird.2. The method according to claim 1, characterized in that in Step a) Diethyl cyanide / triethylamine is used. 3. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß in Schritt b) Pentafluorphenol/Dicyclo­ hexylcarbodiimid eingesetzt wird.3. The method according to any one of the preceding claims, characterized characterized in that in step b) pentafluorophenol / dicyclo hexylcarbodiimide is used. 4. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß in Schritt c) zunächst Pentafluorphenol- Trifluoracetat und dann Triethylamin/Pd/C, H2 eingesetzt wird.4. The method according to any one of the preceding claims, characterized in that in step c) first pentafluorophenol trifluoroacetate and then triethylamine / Pd / C, H 2 is used. 5. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß in Schritt d) zunächst die Ketogruppe reduziert und dann das 1'-C-Atom acetyliert wird.5. The method according to any one of the preceding claims, characterized characterized in that in step d) the keto group is reduced and then the 1'-C atom is acetylated. 6. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß in Schritt f) Tetrabutylammoniumfluorid eingesetzt wird.6. The method according to any one of the preceding claims, characterized characterized in that in step f) tetrabutylammonium fluoride is used. 7. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß R eine C1-6-Alkylgruppe, eine Arylgruppe mit bis zu vier gegebenenfalls annelierten Ringen oder eine Heteroarylgruppe mit bis zu vier gegebenenfalls annelierten Ringen ist, wobei jeder Ring bis zu drei Heteroatome wie z. B. N, O oder S-Atome enthalten kann.7. The method according to any one of the preceding claims, characterized characterized in that R is a C1-6 alkyl group, an aryl group with up to four if necessary  fused rings or a heteroaryl group with up to four optionally fused rings, each ring up to three heteroatoms such as B. can contain N, O or S atoms. 8. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß daß R eine Ethyl-, Propyl-, i-Butyl, sec- Butyl oder tert.-Butylgruppe ist.8. The method according to any one of the preceding claims, characterized characterized in that R is ethyl, propyl, i-butyl, sec- Is butyl or tert-butyl group. 9. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß die Verbindung der Formel 3 als Racemat eingesetzt wird.9. The method according to any one of the preceding claims, characterized characterized in that the compound of formula 3 as a racemate is used. 10. Verfahren nach einem der Ansprüche 1 bis 8, dadurch gekenn­ zeichnet, daß die Verbindung der Formel 3 als L-Enantiomeres eingesetzt wird.10. The method according to any one of claims 1 to 8, characterized records that the compound of formula 3 as an L-enantiomer is used. 11. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, daß die Verbindung der Formel 26 als Diastereomerengemisch eingesetzt wird.11. The method according to any one of the preceding claims, characterized characterized in that the compound of formula 26 as Diastereomer mixture is used. 12. Verfahren nach einem der Ansprüche 1 bis 10, dadurch gekennzeichnet, daß die Verbindung der Formel 26 in der allo- Form eingesetzt wird.12. The method according to any one of claims 1 to 10, characterized characterized in that the compound of formula 26 in the allo- Form is used. 13. Verfahren nach einem der vorstehenden Ansprüche, dadurch ge­ kennzeichnet, daß die Verbindung der Formel 22 als Diastereo­ merengemisch eingesetzt wird.13. The method according to any one of the preceding claims, characterized ge indicates that the compound of formula 22 as a diastereo mixed mixture is used. 14. Verfahren nach einem der Ansprüche 1 bis 12, dadurch ge­ kennzeichnet, daß die Verbindung der Formel 22 diastereo­ merenrein eingesetzt wird.14. The method according to any one of claims 1 to 12, characterized ge indicates that the compound of formula 22 diastereo is used purely. 15. Verbindung der Formel 9a/b
15. Compound of formula 9a / b
16. Verbindung nach Anspruch 16 mit einer R,R-, S,S-, R,S- oder S,R-Konfiguration.16. A compound according to claim 16 with an R, R, S, S, R, S or S, R configuration. 17. Verbindung der Formel 8
dadurch gekennzeichnet, daß sie als L-Form oder als Racemat vorliegt.17. Compound of formula 8
characterized in that it is present as an L-form or as a racemate. 18. Verfahren zur Herstellung einer Verbindung nach Anspruch 15 oder 16, dadurch gekennzeichnet, daß eine Verbindung der Formel 8 - vorzugsweise mit Natriumborhydrid in einem Alkohol - reduziert wird.18. A method for producing a compound according to claim 15 or 16, characterized in that a compound of the formula 8 - preferably with sodium borohydride in an alcohol - is reduced. 19. Verbindung der Formel 22
worin R1 ein H-Atom, eine Alkylgruppe, eine Arylgruppe oder eine Heteroarylgruppe ist. 19. Compound of Formula 22
wherein R 1 is an H atom, an alkyl group, an aryl group or a heteroaryl group. 20. Verfahren zur Herstellung einer Verbindung nach Anspruch 19, dadurch gekennzeichnet, daß eine Verbindung der Formel 8
  • a) gegebenenfalls mit NaOH in H2O und
  • b) gegebenenfalls mit TMSEOH und DCC versetzt wird und
  • c) gegebenenfalls in Gegenwart von NaH mit einer Verbindung der Formel 21
    umgesetzt und dann
  • d) gegebenenfalls mit Tetrafluorbutylammoniumfluorid versetzt wird.
20. A method for producing a compound according to claim 19, characterized in that a compound of formula 8
  • a) optionally with NaOH in H 2 O and
  • b) TMSEOH and DCC are optionally added, and
  • c) optionally in the presence of NaH with a compound of formula 21
    implemented and then
  • d) optionally with tetrafluorobutylammonium fluoride.
21. Verfahren zur Herstellung einer Verbindung der Formel 8 dadurch gekennzeichnet, daß eine Verbindung der Formel 7
vorzugsweise in Gegenwart einer sauren Verbindung hydrolysiert wird.21. A process for the preparation of a compound of formula 8, characterized in that a compound of formula 7
is preferably hydrolyzed in the presence of an acidic compound. 22. Verfahren zur Herstellung einer Verbindung der Formel 7 dadurch gekennzeichnet, daß eine Verbindung der Formel 5
oxidiert und mit einer Verbindung der Formel 6
umgesetzt wird.22. A process for the preparation of a compound of formula 7, characterized in that a compound of formula 5
oxidized and with a compound of formula 6
is implemented. 23. Verbindung der Formel 15b
23. Compound of Formula 15b
24. Verfahren zur Herstellung einer Verbindung der Formel 15b dadurch gekennzeichnet, daß eine Verbindung der Formel 14
einer Ringöffnung unterworfen und das entstandene Produkt verestert wird.24. A process for the preparation of a compound of formula 15b, characterized in that a compound of formula 14
subjected to a ring opening and the resulting product is esterified. 25. Verfahren nach Anspruch 24, dadurch gekennzeichnet, daß die Verbindung 15b diastereomerenrein ist.25. The method according to claim 24, characterized in that the Compound 15b is diastereomerically pure. 26. Verfahren zur Herstellung einer Verbindung der Formel 14 dadurch gekennzeichnet, daß eine Verbindung der Formel 12
hydriert wird.26. A process for the preparation of a compound of formula 14, characterized in that a compound of formula 12
is hydrogenated. 27. Verfahren zur Herstellung einer Verbindung der Formel 12 dadurch gekennzeichnet, daß eine Verbindung der Formel 11
oxidiert und dann um eine C3-Einheit verlängert wurde.27. A process for the preparation of a compound of formula 12, characterized in that a compound of formula 11
oxidized and then extended by a C 3 unit. 28. Verfahren nach Anspruch 27, dadurch gekennzeichnet, daß die Oxidation eine Swern-Oxidation ist.28. The method according to claim 27, characterized in that the Oxidation is a Swern oxidation. 29. Verfahren nach einem der Ansprüche 27 oder 28, dadurch gekennzeichnet, daß die Verlängerung um eine C3-Einheit durch eine Wittigreaktion erfolgt.29. The method according to any one of claims 27 or 28, characterized in that the extension by a C 3 unit is carried out by a Wittig reaction. 30. Verbindung der Formel 25
30. Compound of formula 25
31. Verfahren zur Herstellung einer Verbindung der Formel 25, dadurch gekennzeichnet, daß eine Verbindung der Formel 24
mit TNSEOH und DCC und dann mit CF3CO2H/CH2Cl2 umgesetzt wird.31. A process for the preparation of a compound of formula 25, characterized in that a compound of formula 24
with TNSEOH and DCC and then with CF 3 CO 2 H / CH 2 Cl 2 . 32. Verfahren zur Herstellung einer Verbindung der Formel 24, dadurch gekennzeichnet, daß eine Verbindung der Formel 23
in 2-Position methyliert wird, wobei ein Evans-Auxiliar eingesetzt werden kann, und die Esterfunktion dann hydrolysiert wird.32. A process for the preparation of a compound of formula 24, characterized in that a compound of formula 23
is methylated in the 2-position, where an Evans auxiliary can be used, and the ester function is then hydrolyzed. 33. Verfahren zur Herstellung einer Verbindung der Formel 23, dadurch gekennzeichnet, daß eine Verbindung der Formel 11
zunächst einer Oxidation unterzogen wird und dann im Rahmen einer Wittig Reaktion umgesetzt wird.33. A process for the preparation of a compound of formula 23, characterized in that a compound of formula 11
is first subjected to an oxidation and then implemented in a Wittig reaction. 34. Verfahren zur Herstellung von einer Verbindung der Formel 3
dadurch gekennzeichnet, daß eine Verbindung der Formel 2
reduktiv aminiert wurde.34. Process for the preparation of a compound of formula 3
characterized in that a compound of formula 2
was reductively aminated. 35. Verfahren nach Anspruch 34, dadurch gekennzeichnet, daß die Verbindungen der Formeln 2 und 3 in L-Form vorliegen.35. The method according to claim 34, characterized in that the Compounds of formulas 2 and 3 are in L-form.
DE10008089A 2000-02-22 2000-02-22 Production of tubulysin compounds comprises multi-stage process including condensation of N-methylpipecolinoyl-isoleucine with substituted thiazole-4-carboxylic acid derivative Withdrawn DE10008089A1 (en)

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