WO2020022892A1 - Dérivés de tubulysine et leurs procédés de préparation - Google Patents

Dérivés de tubulysine et leurs procédés de préparation Download PDF

Info

Publication number
WO2020022892A1
WO2020022892A1 PCT/NL2019/050481 NL2019050481W WO2020022892A1 WO 2020022892 A1 WO2020022892 A1 WO 2020022892A1 NL 2019050481 W NL2019050481 W NL 2019050481W WO 2020022892 A1 WO2020022892 A1 WO 2020022892A1
Authority
WO
WIPO (PCT)
Prior art keywords
substituted
unsubstituted
benzyl
general formula
compound
Prior art date
Application number
PCT/NL2019/050481
Other languages
English (en)
Inventor
Alexander Stephan Siegfried DÖMLING
Sayed Kamel GODA
Original Assignee
Rijksuniversiteit Groningen
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rijksuniversiteit Groningen filed Critical Rijksuniversiteit Groningen
Publication of WO2020022892A1 publication Critical patent/WO2020022892A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/021—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)n-C(=0)-, n being 5 or 6; for n > 6, classification in C07K5/06 - C07K5/10, according to the moiety having normal peptide bonds

Definitions

  • the invention relates to the field of medicinal chemistry.
  • it relates to novel means and methods for the synthesis of tubulysin and derivatives thereof, for use as cytotoxic agents e.g. in targeted drug delivery.
  • Antibody-drug conjugates or ADCs are an important class of highly potent biopharmaceutical drugs designed as a targeted therapy for the treatment of cancer. Unlike chemotherapy, ADCs are intended to target and kill only the cancer cells and spare healthy cells. ADCs are complex molecules composed of an antibody linked to a biologically active cytotoxic (anticancer) payload or drug. Antibody-drug conjugates are examples of bioconjugates and immunoconjugates. By combining the unique targeting capabilities of monoclonal antibodies with the cancer-killing ability of cytotoxic drugs, antibody-drug conjugates are designed to allow sensitive discrimination between healthy and diseased tissue. This means that, in contrast to traditional chemotherapeutic agents, antibody-drug conjugates should selectively target and attack the cancer cell so that healthy cells are less severely affected.
  • a stable link between the antibody and cytotoxic agent is a crucial aspect of an ADC.
  • a highly stable ADC linker will ensure that less of the cytotoxic payload falls off in circulation, driving an improved safety profile, and will also ensure that more of the payload arrives at the cancer cell, driving enhanced efficacy.
  • Linkers are based on chemical motifs including disulfides, hydrazones or peptides (cleavable), or thioethers (noncleavable) and control the distribution and delivery of the cytotoxic agent to the target cell.
  • Highly toxic small molecules or natural products have recently found important commercial applications in ADCs.
  • dolastatine, maytansin and other natural product derivatives are coupled to antibodies to increase their efficacy.
  • Several of such compounds are marketed already and multiple are in chnical trials. (Maturing antibody-drug conjugate pipeline hits 30 Asher Mullard, Nature Reviews Drug Discovery 12, 329- 332 (2013) doi: 10.1038/nrd4009).
  • myxobacterial cultures are exceptionally potent cell-growth inhibitors that act by inhibiting tubulin polymerisation and thereby induce apoptosis. See Sasse et al. J. Antibiot. 2000, 53, 879-885; W098/13375. These compounds show high cytotoxicity in the low picomolar ICr,o in a panel of cancer cell lines and are thus of interest as potential anticancer therapeutics.
  • Tubulysins (I) are tetrapep tides, containing three unusual amino acids; thus, the total synthesis poses a considerable challenge to organic chemists.
  • tubulysins and to enhance their selectivity, polymer conjugates and bioconjugates of tubulysins were developed (see e.g. W02004/005326) which exhibit a higher selectivity by a given cytotoxicity as well as lower toxicity as compared to the unconjugated compounds.
  • W02004/005326 a polymer conjugates and bioconjugates of tubulysins
  • predominantly cancer cells are targeted in the human and the animal body and healthy tissue is not affected.
  • tubulysin derivatives and (antibody) conjugates are very promising.
  • tubulysin derivatives thus obtained are super potent and can be attached towards biological matter (e.g. mAbs) through at least 3 different linker positions. Furthermore, the synthetic method offers full stereo-control. The new approach allows to fine-tune the properties of tubulysin ADCs in a much better way than offered by current products.
  • the invention provides method for preparing a tubulysin derivative, comprising reacting compounds A, B and C in a 3-component Passerini reaction,
  • compound A is a carboxyhc acid according to the general formula A
  • Ri represents a substituted or unsubstituted alkyl; a substituted or unsubstituted cycloalkyl; or a substituted or unsubstituted benzyl,
  • R2 represents H, a substituted or unsubstituted alkyl, or a substituted or unsubstituted cycloalkyl
  • R.3 represents a substituted or unsubstituted alkyl; a substituted or unsubstituted cycloalkyl; or a substituted or unsubstituted benzyl;
  • R4 represents H, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted benzyl;
  • Rr represents a substituted or unsubstituted alkyl or a substituted or unsubstituted cycloalkyl; preferably a substituted or unsubstituted cycloalkyl; or wherein R i and Rr, are connected to form a 4 to 7 membered ring;
  • Re represents a substituted or unsubstituted alkyl; a substituted or unsubstituted cycloalkyl; a substituted or unsubstituted benzyl, or COOR’, where R’ is an optionally substituted alkyl, cycloalkyl, benzyl or aroyl moiety; wherein compound B is an aldehyde according to the general formula B
  • R 7 , Re, R 3 ⁇ 4 Rio and Rn each independently represent H
  • F a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl or a substituted or unsubstituted benzyl;
  • Pg 1 is an amine protecting group, preferably a carbamate, a substituted or an unsubstituted benzyl, or a substituted or an unsubstituted sulfonamide; wherein compound C is an isocyanide according to the general formula C
  • R12 represents a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, or a substituted or unsubstituted benzyl;
  • Ri3 represents H, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, or a substituted or unsubstituted benzyl;
  • X represents O, S, Se, or -NH-, preferably S;
  • Pg- represents an X- protecting group, preferably selected from trityl, tert -butyl, adamantyl and substituted benzyl, more preferably tiityl or tert-butyl.
  • a moiety may be substituted, such as by use of “unsubstituted or substituted” or“optionally substituted” phrasing as in “unsubstituted or substituted alkyl” or“optionally substituted benzyl,” such moiety may have one or more independently selected substituents, preferably one to five in number, more preferably one or two in number. Substituents and substitution patterns can be selected by one of ordinary skill in the art, having regard for the moiety to which the substituent is attached, to provide compounds that are chemically stable and that can be synthesized by techniques known in the art as well as the methods set forth herein.
  • substituted refers to groups in which one or more hydrogen atoms are replaced with one or more moieties selected from the group consisting of hydroxyl, amino, alkylamino, arylamino, alkoxy , aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate and phosphonate.
  • One or more of the hydrogen atoms attached to carbon atom may be replaced by one or more halogen atoms, e.g. fluorine or chlorine or both, such as trifluoromethyl, difluoromethyl, fluorochloroniethyl.
  • This expression further refers to groups which are exclusively or ⁇ additionally replaced with unsubstituted Ci-Co alkyl, C2 -C G alkenyl, C2 -C G alkynyl, Ci -C G heteroalkyl, C;; -C10 cycloalkyl, C2 -C9 heterocycloalkyl, C G - Cio aryl, C 1 -Cg heteroaryl, C ? -C12 aralkyl or C2 -Cit heteroaralkyl groups.
  • substituted alkyl refers to an alkyl, cycloalkyl or benzyl that is
  • substituents preferably 1 or 2 substituents, selected from the group consisting of hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, and phosphonate.
  • one or more e.g. 1- 5, 1-3 or 1, of the hydrogen atoms attached to carbon atom of the alkyl, cycloalkyl or benzyl is replaced by one or more halogen atoms, e.g. fluorine or chlorine or both, such as trifluoromethyl, difluoromethyl,
  • alkyl refers to a saturated straight or branched hydrocarbon chain of typically Ci to Cio, and specifically includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, cyclohexylm ethyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl, and the like.
  • the alkyl is a lower alkyl.
  • alkyl when alkyl is a suitable moiety, lower alkyl is preferred. Similarly, when alkyl or lower alkyl is a suitable moiety, unsubstituted alkyl or lower alkyl is preferred.
  • Alkyl groups can be optionally substituted with one or more moieties selected from the group consisting of hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate.
  • One or more of the hydrogen atoms attached to carbon atom on alkyl may be replaces by one or more halogen atoms, e.g. fluorine or chlorine or both, such as trifluorom ethyl,
  • cycloalkyl refers to a saturated or partially unsaturated (e.g. cycloalkenyl) cyclic group containing one or more rings (preferably 1 or 2), the total of 3 to 14 ring carbon atoms, preferably 3 to 10 (especially 3 containing 4, 5, 6 or 7) ring carbon atoms. In one embodiment, it refers to a saturated hydrocarbon ring having 3-8 carbon atoms, preferably, 3-6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
  • the cycloalkyl group may also be substituted on the ring by and alkyl group, such as cyclopropylmethyl and the like.
  • the term“substituted benzyl” refers to a benzyl radical which is substituted by fluorine, chlorine, bromine, nitro, Ci-C i alkyl, C1-C4- halogenoalkyl or C 1 -Ui alkoxy.
  • the benzyl is substituted with chlorine, bromine, nitro, methyl or methoxy.
  • stereoisomers are specifically indicated (e.g., by a bolded or dashed bond at a relevant stereocenter in a structural formula, by depiction of a double bond as having E or Z configuration in a structural formula, or by use stereochemistry-designating nomenclature), all stereoisomers are included within the scope of the invention, as pure compounds as well as mixtures thereof. Unless otherwise indicated, individual enantiomers, diastereomers, geometrical isomers, and
  • the invention uses a carboxylic acid compound A of the general formula A, wherein Rt represents isopropyl, tert-butyl, iso-butyl, sec-butyl, cyclopropylmethyl or cyclobutylmethyl;R2 is H; R3 is -(CFbhi- CHa, wherein n is 3, 4 or 5; R 4 and R5 are connected to form a 4 to 7 membered ring; or Rais a substituted or unsubstituted cycloalkyl; and/or Re is selected from the group consisting of benzyloxycarbonyl, 4- azidobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2- nitr ob enzyloxy c arb onyl , 4,5- dimethoxy -2 -nitr ob enzyloxy c arb onyl , 3,5- dimethoxybenzyloxycarbonyl, 1-nap
  • compound A is of the formula A’ wherein R 4 and R ⁇ , are connected, and together form a cyclic structure comprising 3, 4, 5, 6 or 7 carbon atoms.
  • Ra represents H or methyl, and /or R ⁇ ; represents H, methyl, ethyl, propyl or cyclopropyl.
  • the method uses an aldehyde compound B according to the general formula B wherein R? is H; Rg is H; R9 is selected from the group consisting of isopropyl, cyclopropyl, cyclobutyl, isobutyl, sec- butyl, tert -butyl and cyclopropylmethyl; Rio is H; and/or Rn is selected from the group consisting of methyl, ethyl, propyl, butyl, isopropyl, cyclopropyl and cyclopropylmethyl.
  • the Pg 1 moiety in compound B is an amine protecting group, preferably a carbamate, a substituted or an unsubstituted benzyl.
  • Pg 1 is selected from the group consisting of benzyloxycarbonyl, 4- azidobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2- nitrobenzyloxycarbonyl, 4,5-dimethoxy-2-nitrobenzyloxycarbonyl, 3,5- dimethoxybenzyloxycarbonyl, 1-naphthylmethoxycarbonyl, 4- acetyloxyb enzyloxy c arb onyl , fluor enyloxy c arb onyl , tert -butyloxyc arb onyl , allyloxycarbonyl, methyl carbamate, ethyl carbamate, benzyl, 4- methoxybenzyl and 3,4-dimethoxybenzyl.
  • Preferred isocyanide compounds C according to the general formula C include those wherein R12 is methyl, ethyl or tert -butyl; R13 is H or methyl, and/or wherein X represents S.
  • the X-protecting group Pg 2 is preferably selected from trityl, tert -butyl, adamantyl and substituted benzyl, more preferably Pg 2 is trityl or tert-butyl.
  • compounds A, B and C can be reacted in any suitable solvent or solvent mixture.
  • a non-coordinating solvent or solvent mixture is used.
  • good results can be obtained with CH 2 CI 2 , CHCI 3 , CCE, benzene, THF, CH 3 CN, 1,4-dioxane, or 1,2-dichloroethane.
  • compounds A, B and C are reacted in a mixture of CE CbiTHF (1: 1 v/v).
  • the reaction is most easily performed at room temperature. However, reaction conditions above or below room temperature are also encompassed.
  • the reaction time can range from a few hours up to a few days.
  • the three starting compounds may be present in the starting reaction mixture about equimolar amounts, for example wherein A : B : C is 0.8- 1.2: 0.8- 1.2: 0.8- 1.2, preferably 0.9-1.1:0.9-1.1:09-1.1.
  • the Passerini reaction is preferably carried out from 1 mmol to 100 mmol scale.
  • reaction products are suitably purified through flash chromatography (e.g. cyclohexane/EtOAc 20:80 v/v).
  • the by-product can also be recovered and coupled with dipeptide acid to form an ester.
  • a method of the invention further comprising isolating the 3-component Passerini reaction product of Formula D
  • a method of the invention advantageously further comprises subjecting the Passerini reaction product of formula D to (a) an acyl migration reaction and (b) a cyclodehydration reaction of the Cys-amide. This will yield a thiazole compound of the general formula E having a hydroxyl moiety
  • the acyl migration reaction is suitably performed in a two-step process involving exposure to diethylamine (DEA) followed by exposure to
  • TSA trimethylamine
  • Cyclodehydration of the Cys-amide may also be performed in a two-step process, preferably involving incubation in the presence of TiCL to obtain a thiazoline-containing compound, followed by oxidation in the presence of Mn0 2 to obtain a thiazole moiety.
  • Activated M11O2 having a pore size of ⁇ 5 microns is preferred.
  • acyl migration reaction and the cyclodehydration reaction are performed are not critical.
  • acyl migration precedes cyclodehydration.
  • cyclodehydration precedes acyl migration.
  • the method comprises
  • a method according to the invention advantageously further comprises hydrolyzing the Ria-containing ester of the general formula E to obtain a carboxylic acid compound of the general formula F
  • carboxylic acid compound of the formula F may be reacted with a compound of the general formula G followed by removal of protecting moiety R 21 , to obtain a compound of the general formula H
  • R14, R15, Rin, R20 and protecting moiety R21 each independently represent H, a substituted or unsubstituted alkyl, a substituted or
  • R17 and Rts each independently represent H, F, a substituted or unsubstituted alkyl, or a substituted or unsubstituted cycloalkyl and Rtg represents H, F, a substituted or unsubstituted alkyl, a substituted or unsubstituted
  • Pg 3 ⁇ 4 of compound G is an amine protecting group, preferably a carbamate, a substituted or unsubstituted benzyl, preferably selected from the group consisting of tertbutyl sulfine, benzyloxycarbonyl, 4-azidobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2 -nitrobenzyloxy carbonyl, 4,5-dimethoxy-2- nitrobenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 1- naphthylmethoxycarbonyl, 4-acetyloxybenzyloxycarbonyl,
  • fluorenyloxycarbonyl tert-butyloxycarbonyl, allyloxycarbonyl, methyl carbamate, ethyl carbamate, benzyl, 4-methoxybenzyl and 3,4- dimethoxyb enzyl .
  • R14 represents H
  • Ri represents H, benzyl, (substituted) benzyl
  • Ri ⁇ represents H, benzyl, substituted benzyl (if Rir > is H) and/or
  • Ri T and Ris represent H or F.
  • Rii and/or R20 are preferably independently selected from the group consisting of methyl, ethyl, propyl, butyl, isopropyl, cyclopropyl and cyclopropylmethyl.
  • R21 is preferably H, methyl or ethyl.
  • the carboxylic acid compound of the formula F is reacted with tubuphenylalanine of the formula
  • a further aspect of the invention relates to a method for providing a tubuphenylalanine of the above formula using (S)-(-)methylsuccinic anhydride as intermediary compound. This method typically only comprising 6 steps, is shorter than the current synthetic procedures used and moreover has a better yield (up to nearly 60%) and increased stereoselectivity.
  • the method comprises the steps of : (i) refluxing (S)-(-)methylsuccinic acid in the presence of acetyl chloride to obtain(S)-(-)methylsuccinic anhydride
  • a method of the invention for providing a tubulysin derivative may further comprise the acylation of the hydroxyl group a compound of the general formula H to obtain an ac(et)ylated compound of the general Formula I
  • R2 1 represent acetyl, acyl (substituted) alkyl, acyl cycloalkyl, or acyl benzyl, preferably acetyl or acyl derivative of methyl, ethyl, tert -butyl or benzyl.
  • a method of the present invention allows for the introduction of a variety of different linker types used in the conjugation of payloads to small molecules, polymers, peptides, proteins, antibodies, antibody fragments etc. can be adopted and thereby, many different conjugation methods can be applied.
  • Spacer systems at different positions can be used either directly for conjugation by using different conjugation technologies such as chemical conjugation methods known in the art, or enzymatic conjugations using transglutaminases, sortases or other enzymes or which can be used in combination with commonly described linker systems known in the art .
  • a method as herein disclosed provides tubulysin derivatives that can be attached toward biological matter, e.g. monoclonal antibodies, through at least 3 different linker positions.
  • the properties of, for example, ADC’s can be fine-tuned in a better way as compared to existing products.
  • the present invention particularly finds it use in the manufacture of a cytotoxic tubulysin derivative or tubulysin prodrug.
  • a method as herein disclosed provides a conjugate comprising a tubulysin compound covalently linked to a targeting moiety that specifically or preferentially binds to a chemical entity on a target cell, which target cell preferably is a cancer cell.
  • the targeting moiety is an antibody— more preferably a monoclonal antibody; even more preferably a human monoclonal antibody— and the chemical entity is a tumor associated antigen.
  • the tumor associated antigen can be one that is displayed on the surface of a cancer cell or one that is secreted by a cancer cell into the surrounding extracellular space.
  • a method of the invention comprising a step of reacting the ac(et)ylated compound of the general Formula I with Glu-(OMe)2 or alternative amino acid ester to obtain a compound of the formula J wherein R22 is H or an amino acid side chain, preferably CH2-CH2-COOZ wherein Z is H or methyl,
  • R23 H or a carboxyl protecting group, in particular methyl, ethyl, or tert- butyl; preferably H, methyl, ethyl or tert -butyl.
  • the method comprises the synthesis of a glutamic acid conjugate of tubulysin, which finds its use in a prodrug approach using glucarpidase or an enzyme showing a similar activity.
  • the method comprises the formation of a tubulysin-prodrug comprising one or more glutamate residue(s) linked to an amidic, urethanic or ureidic bond.
  • the invention provides one of the following exemplary compounds
  • Fmoc-6-Val 3 (1.0 mmol) was suspended in 20 mL of toluene, and
  • reaction was cooled to room temperature and filtered through a medium glass frit.
  • the filter cake was washed with 3 x 2 mL of ethyl acetate, and the combined filtrates were concentrated and purified by flash column chromatography to yield aldehyde 7 as a yellow gum.
  • the ketoacid 18 (7.0 mmol) was dissolved in EtOH (40 mL), concentrated H2SO4 (1.1 mL, 21.0 mmol) was added at room temperature, and then the reaction mixture was refluxed overnight. After evaporation of the solvent, H2O (10 mL) was added, and the mixture was neutralized with a 2 M aqueous NaOH solution and extracted with EtOAc (3 x 15 mL). The combined organic phases were dried (NasSO i). After filtration and evaporation of the solvent, the expected ketoester obtained as oil.
  • Ketone (1.0 mmol) was added to a solution of 20 (1.5 mmol) and Ti(OEt) i (3.0 mmol) in THF (15 mL) at room temperature (rt). The reaction mixture was heated at 70 °C for 8h and the reaction conversion was followed by TLC. Once the reaction was determined to be complete by TLC, the mixture was cooled to room temperature and then to -78 °C. L-Selectride (1 M solution in THF) was added dropwise. The reaction mixture was stirred at same temperature for 3 h. Once the reduction was determined to be complete by TLC, the reaction mixture was quenched by dropwise addition of MeOH at 0 °C until gas evolution was no longer observed.
  • the crude reaction mixture was poured into an equal volume of brine while being rapidly stirred.
  • the resulting suspension was filtered through a plug of Cellite, and the filter cake was washed with EtOAc.
  • the filtrate was washed with brine, and the brine layer was extracted with EtOAc (3x).
  • the combined organic portions were dried (Na 2 S0 4 ), filtered, and concentrated.
  • the product 21 was purified by silica gel chromatography (hexanes/EtOAc).
  • Acid 27 (1.0 mmol) was added to a 0.2 M solution of pentafluorophenol (1.25 mmol) and DIC (1.0 mmol) in CH 2 C1 2 at 0 °C. The reaction mixture was warmed to rt, stirred for 24 h, and concentrated under reduced pressure. EtOAc (10 mL) was added, and the crude product was filtered, with rinsing of the reaction vessel with EtOAc. The filtrate was concentrated under reduced pressure, and the crude material was used without further purification.
  • the compound 29 (1.0 mmol) was dissolved in a mixture of MeOH (20 ml). Paraformaldehyde (300 mg, 10 mmol) and 20% Pd/C (106 mg, 0.1 mmol Pd) were added. The reaction mixture was stirred under hydrogen atmosphere for 16 h and afterwards filtered through Cellite. The solvent was then removed under reduced pressure. The product was purified through column chromatography.
  • Acid 30 (1.0 mmol) was added to a 0.2 M solution of pentafluorophenol (1.25 mmol) and DIC (1.0 mmol) in a CH2CI2 at 0 °C. The reaction mixture was warmed to rt, stirred for 24 h, and concentrated under reduced pressure and dissolved in DMF (0.5 M). In another reaction vessel hydrochloride salt of H- Glu-(OMe)2 (3.0 mmol) and diisopropylethylamine (5.0 mmol) were dissolved in DMF (0.335 mL, 0.25 M) and this solution is added to the above prepared pent aflur ophenyl ester.
  • Acid 30 (1.0 mmol) was added to a 0.2 M solution of pentafluorophenol (1.25 mmol) and DIC (1.0 mmol) in a CH2CI2 at 0 °C. The reaction mixture was warmed to rt, stirred for 24 h, and concentrated under reduced pressure and dissolved in acetonitrile (0.5 M).
  • H-Asp-OH, (2.0 mmol) and sodium bicarbonate (4.0 mmole) were dissolved in 2 ml. of water and adjust the pH about 8. Then a solution of pentafluorophenyl ester prepared above (1.0 mmole) 3 mL of acetonitrile was added at room temperature.
  • compound 30 (100 nM test concentration) is highly toxic to HeLa cells after 18hrs treatment, thus demonstrating its suitabihty as cytotoxic agent e.g. in targeted drug delivery.
  • Tup Tubuphenylalanine
  • tubulysin“derivative” is therefore considered in the broad context and allows for:
  • R3 wide range of groups can be incorporated

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne de nouveaux moyens et procédés pour la synthèse de tubulysine et de ses dérivés, qui trouvent leur utilisation, par exemple, en tant qu'agents cytotoxiques dans l'administration ciblée de médicaments. L'invention concerne un procédé de préparation d'un dérivé de tubulysine, qui comprend les étapes consistant à faire réagir des composés A, B et C dans une réaction de Passerini à 3 composants, le composé A étant un acide carboxylique selon la formule générale (A) ; le composé B étant un aldéhyde selon la formule générale (B) ; et le composé C étant un isocyanure selon la formule générale (C).
PCT/NL2019/050481 2018-07-24 2019-07-24 Dérivés de tubulysine et leurs procédés de préparation WO2020022892A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP18185173 2018-07-24
EP18185173.4 2018-07-24

Publications (1)

Publication Number Publication Date
WO2020022892A1 true WO2020022892A1 (fr) 2020-01-30

Family

ID=63041870

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL2019/050481 WO2020022892A1 (fr) 2018-07-24 2019-07-24 Dérivés de tubulysine et leurs procédés de préparation

Country Status (1)

Country Link
WO (1) WO2020022892A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111454230A (zh) * 2020-04-26 2020-07-28 深圳市老年医学研究所 一种天然抗癌药物Tubulysins的关键中间体Tuv的合成方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998013375A1 (fr) 1996-09-23 1998-04-02 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Composes a effet antimycotique et cytostatique, leur procede de production, agents contenant ces composes et dsm 11 092
WO2004005326A2 (fr) 2002-07-09 2004-01-15 Morphochem Aktiengellschaft Fu Conjugues de tubulysine
EP2409983A1 (fr) * 2010-07-19 2012-01-25 Leibniz-Institut für Pflanzenbiochemie (IPB) Analogues de la tubulysine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998013375A1 (fr) 1996-09-23 1998-04-02 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Composes a effet antimycotique et cytostatique, leur procede de production, agents contenant ces composes et dsm 11 092
WO2004005326A2 (fr) 2002-07-09 2004-01-15 Morphochem Aktiengellschaft Fu Conjugues de tubulysine
EP2409983A1 (fr) * 2010-07-19 2012-01-25 Leibniz-Institut für Pflanzenbiochemie (IPB) Analogues de la tubulysine

Non-Patent Citations (18)

* Cited by examiner, † Cited by third party
Title
ASHER MULLARD, NATURE REVIEWS DRUG DISCOVERY, vol. 12, 2013, pages 329 - 332
FAURE ET AL., ORG. LETT., vol. 11, no. 5, 2009, pages 1167 - 1170
J. MED. CHEM., vol. 50, no. 18, 2007, pages 4261 - 4264
J. ORG. CHEM., vol. 48, 1983, pages 77 - 81
J. ORG. CHEM., vol. 66, 2001, pages 8454 - 8462
J. ORG. CHEM., vol. 72, 2007, pages 626 - 629
J. ORG. CHEM., vol. 78, 2013, pages 3647
ORG. LETT., vol. 2, 2000, pages 3289
ORG. LETT., vol. 4, 2002, pages 3001 - 3003
ORG. SYNTH., vol. 12, 1932, pages 66
REF: ORG. LETT., vol. 11, no. 5, 2009, pages 1167 - 1170
SASSE ET AL., J. ANTIBIOT., vol. 53, 2000, pages 879 - 885
SYN. COMMUN., vol. 33, 2003, pages 3089 - 3096
TAKUYA FUKUDA ET AL: "Anti-barnacle Activity of Isocyanides Derived from Amino Acids", CHEMISTRY & BIODIVERSITY, vol. 13, no. 11, 1 November 2016 (2016-11-01), CH, pages 1502 - 1510, XP055544849, ISSN: 1612-1872, DOI: 10.1002/cbdv.201600063 *
TETRAHEDRON LETTERS, vol. 45, 2004, pages 8603 - 8606
THIMMALAPURA M. VISHWANATHA ET AL: "Cysteine Isocyanide in Multicomponent Reaction: Synthesis of Peptido-Mimetic 1,3-Azoles", JOURNAL OF ORGANIC CHEMISTRY, vol. 82, no. 18, 25 August 2017 (2017-08-25), pages 9585 - 9594, XP055544660, ISSN: 0022-3263, DOI: 10.1021/acs.joc.7b01615 *
VISHWANATHA ET AL., J. ORG. CHEM., vol. 82, 2017, pages 9585 - 9594
XUECHEN LI ET AL: "On the Two-Component Microwave-Mediated Reaction of Isonitriles with Carboxylic Acids: Regarding Alleged Formimidate Carboxylate Mixed Anhydrides", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 130, no. 40, 8 October 2008 (2008-10-08), pages 13222 - 13224, XP055544845, ISSN: 0002-7863, DOI: 10.1021/ja8047078 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111454230A (zh) * 2020-04-26 2020-07-28 深圳市老年医学研究所 一种天然抗癌药物Tubulysins的关键中间体Tuv的合成方法

Similar Documents

Publication Publication Date Title
AU714147B2 (en) Inhibitors of farnesyl protein transferase
ES2888800T3 (es) Procedimiento mejorado para la preparación de carfilzomib o sales farmacéuticamente aceptables del mismo
AU2012242122A1 (en) Branched 3-phenylpropionic acid derivatives and the use thereof
US6235717B1 (en) Pharmaceutical compounds
WO1995009864A1 (fr) Nouveau derive peptidique
TW200538450A (en) HCV NS-3 serine protease inhibitors
WO1995003277A1 (fr) Nouveau derive de pyrrolidine
Kurokawa et al. Synthetic studies on antifungal cyclic peptides, echinocandins. Stereoselective total synthesis of echinocandin D via a novel peptide coupling
CA2885973C (fr) Derive de dolastatine-10, procede pour le produire et composition de medicament anticancereux le contenant
WO2000043000A1 (fr) Inhibiteurs de proteasome
US20150299250A1 (en) Macrocyclic compounds and uses thereof
WO2017070878A1 (fr) Dérivés d'acides aminés estérifiés à fonction c14-hydroxyle de triptolide, procédé pour leur préparation et utilisation correspondante
US5504189A (en) Peptides, their preparation and use
CA2702605C (fr) Aldehydes amines non proteges et leurs applications
EP1806141B1 (fr) Antagonistes des récepteurs de type par-2
WO2020022892A1 (fr) Dérivés de tubulysine et leurs procédés de préparation
CN104587487B (zh) 一种应用于靶向给药系统的新的支链连接体
EA014874B1 (ru) Соединения для ингибирования апоптоза
JP3163391B2 (ja) スチルベン誘導体及びそれを含有する制癌剤
TWI816708B (zh) 用於合成環狀縮肽的方法
CN109134511B (zh) C19位氟代的Largazole类似物、其制备方法和用途
EP2864321A1 (fr) Synthèse de télaprévir et de bocéprévir, ou de sels ou de solvates pharmaceutiquement acceptables de ceux-ci et produits intermédiaires correspondants, notamment des acides ss-aminés, préparés par addition aldolique de mukaiyama
JP2003505397A (ja) プソイドマイシンのｎ−アシル側鎖アナログ
CN109134295B (zh) 蒽二酮衍生物及其制备方法和应用
WO2023274313A1 (fr) Dérivé semi-synthétique d'amphotéricine b, son procédé de préparation et son utilisation

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19755991

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19755991

Country of ref document: EP

Kind code of ref document: A1