WO2010016011A1 - Organosulfur compounds, a method of making organosulfur compounds and their use for inhibiting the growth of tumour cells - Google Patents

Organosulfur compounds, a method of making organosulfur compounds and their use for inhibiting the growth of tumour cells Download PDF

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WO2010016011A1
WO2010016011A1 PCT/IB2009/053399 IB2009053399W WO2010016011A1 WO 2010016011 A1 WO2010016011 A1 WO 2010016011A1 IB 2009053399 W IB2009053399 W IB 2009053399W WO 2010016011 A1 WO2010016011 A1 WO 2010016011A1
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compound
formula
para
cancer
oxide
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PCT/IB2009/053399
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French (fr)
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Catherine Hart Kaschula
Roger Hunter
Mohamed Iqbal Parker
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University Of Cape Town
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Priority to US13/057,960 priority Critical patent/US8524781B2/en
Priority to CN2009801394718A priority patent/CN102171184A/en
Priority to EP09804634A priority patent/EP2321271A4/en
Publication of WO2010016011A1 publication Critical patent/WO2010016011A1/en
Priority to ZA2011/01690A priority patent/ZA201101690B/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/095—Sulfur, selenium, or tellurium compounds, e.g. thiols
    • A61K31/105—Persulfides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/18—Sulfonamides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/4035—Isoindoles, e.g. phthalimide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/64—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and sulfur atoms, not being part of thio groups, bound to the same carbon skeleton
    • C07C323/65—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and sulfur atoms, not being part of thio groups, bound to the same carbon skeleton containing sulfur atoms of sulfone or sulfoxide groups bound to the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/44—Iso-indoles; Hydrogenated iso-indoles
    • C07D209/48—Iso-indoles; Hydrogenated iso-indoles with oxygen atoms in positions 1 and 3, e.g. phthalimide

Definitions

  • ORGANOSULFUR COMPOUNDS A METHOD OF MAKING ORGANOSULFUR COMPOUNDS AND THEIR USE FOR INHIBITING THE GROWTH OF TUMOUR
  • This invention relates to new compounds, to compounds useful for the inhibition of the growth of tumour cells, to a new process for the synthesis of said compounds, to the use of the compounds in the preparation of medicaments for the inhibition of the growth of tumour cells, and to methods for the inhibition of the growth of tumour cells.
  • Garlic ⁇ allium sativum dietary supplements have a demonstrated ability to reduce the risk of cancer in human beings.
  • the potential chemo-preventative effect of garlic has in the past been the subject of various clinical trials. The outcomes of these trials were contradictory depending on the type of tumour examined and the garlic preparation used. This is due to the fact that crude extracts of garlic contain numerous organosulfur compounds with varying stability and biological activity.
  • organosulfur compounds present in garlic have been well characterized. These compounds include allyl disulfides, allyl thiosulfinates and cysteine sulfoxides. Upon maceration of the garlic bulb one of these organosulfides, S- allylcysteine-S-oxide (alliin), is converted to 2-propenethiosulfinate (allicin) by the enzyme allinase.
  • Ajoene has been shown to offer strong protection against TPA-promoted carcinogenesis on mouse skin, and to strongly inhibit metastasis to the lungs in the B16/BL6 melanoma tumour model in C57BL/6 mice.
  • topical application of ajoene to the tumours of a group of human patients with either nodular or superficial basal cell carcinoma caused a reduction in tumour size in a large percentage of subjects.
  • Ajoene is able to induce apoptosis in a number of tumour cell lines including human breast, bladder, colorectal, hepatic, prostate, lymphoma, leukemia and skin.
  • Apoptosis is a form of physiological cell death characterized by chromatin condensation, cytoplasmic blebbing, and DNA fragmentation.
  • Two major pathways mediating drug-induced apoptosis have been characterized. One involves the triggering of cell surface death receptors and the other the targeting of mitochondria without the involvement of a receptor / ligand system. It is hypothesized that ajoene induces apoptosis via the latter pathway.
  • Ajoene has been shown to induce apoptosis and arrest HL60 leukemic cells in the G 2 / M phase of the cell cycle in a dose-dependent manner.
  • Ajoene-treated leukemia cells undergo a time-dependent reduction in the anti-apoptotic Bcl-2 protein, resulting in the release of cytochrome C and activation of caspase 3.
  • Ajoene has also been shown to decrease the expression of ⁇ 4 ⁇ 1 integrin in murine melanoma cells, and to induce complete disassembly of the microtubule network in HL60 cells.
  • R 3 is selected from H, COR 9 , para-methoxybenzyl, and trialkylsilyl, in which Rg is alkyl or substituted alkyl; R 4 and R 5 are independently alkyl or R 4 and R 5 together form a phthalimido group;
  • R 6 is alkyl or substituted alkyl
  • R 7 and R 8 are independently alkyl or substituted alkyl.
  • the alkyl groups may be independently selected from methyl, ethyl, propyl, butyl, isopropyl and isobutyl.
  • the alkenyl groups may be independently selected from prop-1-enyl (allyl), 1- propenyl, 1-butenyl, 2-butenyl, 3-butenyl and 1-methyl-2-butenyl.
  • R 1 may be propyl, 3-hydroxypropyl, 3-phthalimidopropyl, ferf-butyl, benzyl, para-methoxybenzyl, o/#7o, para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl, dansyl or 3-(ferf-dimethylsilyloxy)propyl.
  • R 2 may be propyl, prop-1-enyl, para-methoxybenzyl, ortho,para- methoxybenzyl, benzyl or para-f luorobenzyl.
  • R 1 may be propyl, 3-hydroxypropyl, 3-phthalimidopropyl, tert- butyl, benzyl, para-methoxybenzyl, orfr/o,para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl or 3-(fen?-dimethylsilyloxy)propyl and R 2 may be prop-1-enyl.
  • R 1 may be para-methoxybenzyl or ortr»o,para-methoxybenzyl and R 2 may be para-methoxybenzyl, ortho, para-methoxybenzyl, benzyl or para- fluorobenzyl.
  • R 1 may be dansyl and R 2 may be propyl.
  • particular compounds of formula (2) may be: (£/Z)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (3),
  • the compound may be for use in a method of killing or inhibiting the growth of tumour cells and/or treating cancer, such as lung cancer, oesophageal cancer, cervical cancer or breast cancer.
  • a pharmaceutical composition comprising a compound described above and a pharmaceutically acceptable carrier.
  • the composition may be for use in killing or inhibiting the growth of tumour cells and/or for treating cancer.
  • the compound of formula (16) may be acylated with thiolacetic acid via a radical mechanism using a radical initiator, such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, e.g. 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN), to initiate the reaction or using palladium (0) coupling of a vinyl halide.
  • a radical initiator such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, e.g. 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN)
  • the thiol may be produced by hydrolysis of the compound of formula (17) in an alcoholic solvent using an alkali metal base, such as sodium or potassium hydroxide.
  • the alcoholic solvent may be methanol or ethanol.
  • the compound of formula (19) may be oxidized by reacting it with an oxidizing agent to produce the compound of formula (2).
  • the oxidizing agent may be m- chloroperoxybenzoic acid (m-CPBA), peroxybenzoic acid or hydrogen peroxide.
  • the method may further include the step of separating the E- and Z-isomers of the compound of formula (2).
  • a compound of formula (2) in the manufacture of a medicament for the inhibition of the growth of tumour cells and/or for the treatment of cancer.
  • a method of inhibiting the growth of tumour cells and/or treating cancer including the step of administering to a person or animal in need of treatment a pharmaceutically effective amount of a compound of formula (2).
  • Ri and R 2 are linear or branched C1- C5 alkyl; linear or branched C1- C5 alkenyl, with the proviso that R 1 is not prop-1-enyl (allyl); substituted linear or branched C1- C5 alkenyl; substituted linear or branched C1- C5 alkyl; in which the substituents are selected from OR 3 ; NR 4 R 5 ; COOR 6 ;
  • R 3 is selected from H, CORg, para-methoxybenzyl and trialkylsilyl, in which R 9 is alkyl or substituted alkyl;
  • R 4 and R 5 are independently alkyl or R 4 and R 5 together form a phthalimido group;
  • R 6 is alkyl or substituted alkyl;
  • R 7 and R 8 are independently alkyl or substituted alkyl.
  • R 1 is not allyl.
  • Preferred alkyl groups include methyl, ethyl, propyl, butyl, isopropyl and isobutyl.
  • Preferred alkenyl groups include prop-1-enyl (allyl), 1-propenyl, 1-butenyl, 2- butenyl, 3-butenyl and 1-methyl-2-butenyl.
  • R 2 can be prop-1-enyl and R 1 can be any one of propyl, 3- hydroxypropyl, 3-phthalimidopropyl, te/t-butyl, benzyl, para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl, 3-(ferf-dimethylsilyloxy)propyl; or R 1 can be para- methoxybenzyl and R 2 can be one of para-methoxybenzyl, benzyl, para-fluorobenzyl; or R-i can be dansyl and R 2 can be propyl. More specific examples of compounds of formula (2) are shown in Table 1. Table 1: Examples of compounds of formula (2)
  • the compounds of the formula (2) can be used for inhibiting the growth of tumour cells. They can therefore be used for treating cancer, such as lung cancer, oesophageal cancer or breast cancer, by administering an effective amount of the compound to a patient in need of treatment.
  • the compound would typically be included in a pharmaceutical composition with a pharmaceutically acceptable carrier.
  • the composition may include a mixture of the E- and Z-isomers of the compound, only the E-isomer or only the Z-isomer.
  • the compounds can be made by the following method:
  • the compound of formula (16) can be prepared by reacting a substituted leaving group, such as a substituted halide of formula (25) (where X is a halide), with thiourea to form a corresponding thiourea salt (25a) and, typically in a one-pot reaction, reacting the salt with a base and with a propargyl halide or tosylate to form a compound of formula (26).
  • a substituted leaving group such as a substituted halide of formula (25) (where X is a halide)
  • thiourea to form a corresponding thiourea salt (25a)
  • reacting the salt with a base and with a propargyl halide or tosylate to form a compound of formula (26).
  • the substituted halide of formula (25) is generally reacted with the thiourea in an aprotic solvent, such as acetonitrile.
  • the thiouronium salt (25a) is generally hydrolysed in an alcoholic solvent such as methanol or ethanol using an alkali metal base such as sodium or potassium hydroxide.
  • Suitable propargyl halides include propargyl bromide, chloride or iodide.
  • the compound of formula (16) can be acylated with thiolacetic acid via a radical mechanism using a radical initiator, such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, such as 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN), to initiate the reaction or using palladium (0) coupling of a vinyl halide.
  • a radical initiator such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, such as 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN)
  • the thiol of (17) can be produced by hydrolysis of (17) in an alcoholic solvent, such as methanol or ethanol, using an alkali metal base, such as sodium or potassium hydroxide.
  • the compound of formula (19) can be oxidized by reaction with an oxidizing agent, such as m-chloroperoxybenzoic acid (m-CPBA), peroxybenzoic acid or hydrogen peroxide, to produce the compound of formula (2).
  • m-CPBA m-chloroperoxybenzoic acid
  • a mixture of the E- and Z-isomers of the compound of formula (2) can be produced by this method.
  • the E- and Z-isomers of the compound of formula (2), in which R 2 is allyl are represented by formula (20.1) and formula (20.2), respectively.
  • the E- and Z-isomers may, in certain cases, be in the form of an inseparable mixture and, in other cases, in the form of a separable mixture. In the cases where the isomers are in the form of a separable mixture, they may be separated. Suitable separation methods include silica gel column chromatography; preparative TLC and high performance liquid chromatography (HPLC).
  • the compounds were evaluated for their in vitro ability to inhibit cell growth of cultured tumour cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
  • TTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
  • WI38 cells which are normal embryonic lung fibroblasts
  • CT-1 cells which are transformed WI38 fibroblasts
  • WHC01 cells which are oesophageal epithelial cancer cells.
  • EPC2 cells which are normal oesophageal epithelial cells.
  • MDA-MB-231 cells which are human breast epithelial cancer cells.
  • MCF12a cells which are normal human breast epithelial cells.
  • WHCO1 oesophageal squamous cell carcinoma cell line WHCO1 originally established from surgical biopsies of primary oesophageal squamous cell carcinomas, a transformed fibroblast cell line CT-1 or human breast epithelial cells MDA-MB-231 were cultured in DMEM containing 10% foetal calf serum and 1% penicillin and streptomycin at 37 0 C in a humidified atmosphere of 5% CO 2 .
  • 3 x 10 3 cells were plated in 96-well plates in 90 ⁇ L DMEM per well.
  • IC 50 Inhibitory concentration of drug used to cause 50% inhibition of cell growth
  • analogue synthesized display superior activity to ajoene.
  • the most active analogue EZ-11 is fifteen times more active than ajoene at inhibiting growth of WHCO1 oesophageal cancer cells in vitro.
  • the four most active drug candidates that display IC 50 activities under 10 ⁇ M are shown below.
  • Formula (27) General structure of the active-class of compounds with R- 1 0-R 13 being electron donating / lipophilic groups.
  • the applicant has developed a synthetic route to access compounds of formula (2), being analogues of ajoene, with varying substituents at the Ri and R 2 positions.
  • the strongest drug candidates synthesized to date are those compounds with either benzyl or para-methoxybenzyl groups at positions R 1 and/or R 2 of formula (2). These compounds display in vitro IC 50 killing activities on WHC01 and MDA cancer cells at drug concentrations under 10 ⁇ M.

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Abstract

Organosulfur compounds of the general formula (2) are described, wherein R1 and R2 are linear or branched C1- C5 alkyl; linear or branched C1- C5 alkenyl with the proviso that R1 is not prop-1-enyl (allyl); substituted linear or branched C1- C5 alkenyl or substituted linear or branched C1- C5 alkyl, in which the substituents are selected from OR3, NR4R5, COOR6, CONR7R8, in which R3 is selected from H, COR9, para- methoxybenzyl and trialkylsilyl, in which R9 is alkyl or substituted alkyl; R4 and R5 are alkyl or R4 and R5 together form a phthalimido group; R6 is alkyl or substituted alkyl; and R7 and R8 are alkyl or substituted alkyl; substituted or unsubstituted aromatic specifically where R1 and R2 are benzyl, para-methoxybenzyl and/or ortho,para- methoxybenzyl and substituted or unsubstituted heteroaromatic. The compounds can be used for inhibiting the growth of tumour cells and for treating cancer. A pharmaceutical composition and a method of preparing the compounds are also described.

Description

ORGANOSULFUR COMPOUNDS, A METHOD OF MAKING ORGANOSULFUR COMPOUNDS AND THEIR USE FOR INHIBITING THE GROWTH OF TUMOUR
CELLS
BACKGROUND OF THE INVENTION
This invention relates to new compounds, to compounds useful for the inhibition of the growth of tumour cells, to a new process for the synthesis of said compounds, to the use of the compounds in the preparation of medicaments for the inhibition of the growth of tumour cells, and to methods for the inhibition of the growth of tumour cells.
Garlic {allium sativum) dietary supplements have a demonstrated ability to reduce the risk of cancer in human beings. The potential chemo-preventative effect of garlic has in the past been the subject of various clinical trials. The outcomes of these trials were contradictory depending on the type of tumour examined and the garlic preparation used. This is due to the fact that crude extracts of garlic contain numerous organosulfur compounds with varying stability and biological activity.
Many of the organosulfur compounds present in garlic have been well characterized. These compounds include allyl disulfides, allyl thiosulfinates and cysteine sulfoxides. Upon maceration of the garlic bulb one of these organosulfides, S- allylcysteine-S-oxide (alliin), is converted to 2-propenethiosulfinate (allicin) by the enzyme allinase. Two molecules of allicin can then combine and rearrange by a thio- Claisen rearrangement to yield an E / Z mixture of 4,5,9-trithiadodeca-1 ,6,11-triene-9- oxide (£-ajoene and Z-ajoene), the structures of which are represented in Formulae 1.1 and 1.2 respectively. Structurally, ajoene contains interesting sulfoxide (S=O) and unusual vinyl disulfide (=S-S) motifs. Synthesis of ajoene can be conducted in vitro in low yield by thermal decomposition of allicin in an acetone-water mixture.
Figure imgf000003_0001
Formula 1.1 (E-ajoene) Formula 1.2 (Z-ajoene)
Ajoene has been shown to offer strong protection against TPA-promoted carcinogenesis on mouse skin, and to strongly inhibit metastasis to the lungs in the B16/BL6 melanoma tumour model in C57BL/6 mice. In one clinical trial topical application of ajoene to the tumours of a group of human patients with either nodular or superficial basal cell carcinoma caused a reduction in tumour size in a large percentage of subjects.
It has also been shown that ajoene is able to induce apoptosis in a number of tumour cell lines including human breast, bladder, colorectal, hepatic, prostate, lymphoma, leukemia and skin. Apoptosis is a form of physiological cell death characterized by chromatin condensation, cytoplasmic blebbing, and DNA fragmentation. Two major pathways mediating drug-induced apoptosis have been characterized. One involves the triggering of cell surface death receptors and the other the targeting of mitochondria without the involvement of a receptor / ligand system. It is hypothesized that ajoene induces apoptosis via the latter pathway. Ajoene has been shown to induce apoptosis and arrest HL60 leukemic cells in the G2 / M phase of the cell cycle in a dose-dependent manner. Ajoene-treated leukemia cells undergo a time- dependent reduction in the anti-apoptotic Bcl-2 protein, resulting in the release of cytochrome C and activation of caspase 3. These results support the hypothesis that ajoene-induced apoptosis in leukemia cells proceeds via a mitochondria-dependent caspase cascade.
Ajoene has also been shown to decrease the expression of α4β1 integrin in murine melanoma cells, and to induce complete disassembly of the microtubule network in HL60 cells. SUMMARY OF THE INVENTION
According to a first embodiment of the invention, there is provided a compound of formula (2)
O O O IΛO
Il *
O
(2) wherein:
Figure imgf000004_0001
linear or branched C1- C5 alkyl; linear or branched C1- C5 alkenyl, with the proviso that R1 is not prop-1-enyl (allyl); substituted linear or branched C1- C5 alkenyl; or substituted linear or branched C1- C5 alkyl; in which the substituents are selected from
OR3; NR4R5; COOR6; CONR7R8; substituted or unsubstituted aromatic; substituted or unsubstituted heteroaromatic, in which
R3 is selected from H, COR9, para-methoxybenzyl, and trialkylsilyl, in which Rg is alkyl or substituted alkyl; R4 and R5 are independently alkyl or R4 and R5 together form a phthalimido group;
R6 is alkyl or substituted alkyl; and
R7 and R8 are independently alkyl or substituted alkyl.
The alkyl groups may be independently selected from methyl, ethyl, propyl, butyl, isopropyl and isobutyl.
The alkenyl groups may be independently selected from prop-1-enyl (allyl), 1- propenyl, 1-butenyl, 2-butenyl, 3-butenyl and 1-methyl-2-butenyl. In particular, R1 may be propyl, 3-hydroxypropyl, 3-phthalimidopropyl, ferf-butyl, benzyl, para-methoxybenzyl, o/#7o, para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl, dansyl or 3-(ferf-dimethylsilyloxy)propyl.
In particular, R2 may be propyl, prop-1-enyl, para-methoxybenzyl, ortho,para- methoxybenzyl, benzyl or para-f luorobenzyl.
More particularly, R1 may be propyl, 3-hydroxypropyl, 3-phthalimidopropyl, tert- butyl, benzyl, para-methoxybenzyl, orfr/o,para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl or 3-(fen?-dimethylsilyloxy)propyl and R2 may be prop-1-enyl.
More particularly, R1 may be para-methoxybenzyl or ortr»o,para-methoxybenzyl and R2 may be para-methoxybenzyl, ortho, para-methoxybenzyl, benzyl or para- fluorobenzyl.
Even more particularly, R1 may be dansyl and R2 may be propyl.
For example, particular compounds of formula (2) may be: (£/Z)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (3),
(EZZ)- 4,8,9-trithiadodeca-6,11-diene-1-ol-4-oxide (4),
(E/Z)-12-phthalimido-4,5,9-trithiadodeca-1,6-diene-9-oxide (5),
(£/Z)-10,10-dimethyl-4,5,9-trithiaundeca-1 ,6-diene-9-oxide (6),
(EZZ)-10-phenyl-4,5,9-trithiadeca-1 ,6-diene-9-oxide (7), (EZZ)- 10-(p-methoxyphenyl)-4,5,9-trithiadeca-1 ,6-diene-9-oxide (8),
(EZZ)-I 2-(p-methoxybenzyloxy)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (9),
(£ZZ)-1-(p-fluorophenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide (10),
(£/Z)-1-(p-methoxyphenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide (11),
(£/Z)-1-phenyl-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide (12), (E/Z)-1-(dansylamino)-4,5,9-trithiadodeca-6-ene-9-oxide (13),
(EZZ)- 4,5,9-trithiadodeca-6-ene-9-oxide (14), or
(EZZ)- 2, 3, 7-trithiadeca-4-ene-7-oxide (15). The compound may be for use in a method of killing or inhibiting the growth of tumour cells and/or treating cancer, such as lung cancer, oesophageal cancer, cervical cancer or breast cancer.
According to a further embodiment of the invention, there is provided a pharmaceutical composition comprising a compound described above and a pharmaceutically acceptable carrier. The composition may be for use in killing or inhibiting the growth of tumour cells and/or for treating cancer.
According to a further embodiment of the invention, there is provided a method of making a compound of formula (2), the method including the steps of:
(i) acylating a compound of formula (16), wherein R1 is as described in claim 1 , with thiolacetic acid to form a thioacetate compound of formula (17)
Figure imgf000006_0001
(16)
Figure imgf000006_0002
(17)
(ii) generating a thiol by treating the compound of formula (17) with a base;
(iii) reacting the thiol with a compound of formula (18), prepared from a tosylate, halide or amide of R2, wherein R2 is as described in claim 1, to produce a compound of formula (19)
Figure imgf000006_0003
(iv) and oxidizing the compound of formula (19) to produce the compound of formula (2)
Figure imgf000006_0004
(2).
The compound of formula (16) may be acylated with thiolacetic acid via a radical mechanism using a radical initiator, such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, e.g. 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN), to initiate the reaction or using palladium (0) coupling of a vinyl halide.
The thiol may be produced by hydrolysis of the compound of formula (17) in an alcoholic solvent using an alkali metal base, such as sodium or potassium hydroxide. The alcoholic solvent may be methanol or ethanol.
The compound of formula (19) may be oxidized by reacting it with an oxidizing agent to produce the compound of formula (2). The oxidizing agent may be m- chloroperoxybenzoic acid (m-CPBA), peroxybenzoic acid or hydrogen peroxide.
The method may further include the step of separating the E- and Z-isomers of the compound of formula (2).
According to a further embodiment of the invention, there is provided the use of a compound of formula (2) in the manufacture of a medicament for the inhibition of the growth of tumour cells and/or for the treatment of cancer.
According to a further embodiment of the invention, there is provided a method of inhibiting the growth of tumour cells and/or treating cancer, the method including the step of administering to a person or animal in need of treatment a pharmaceutically effective amount of a compound of formula (2).
DETAILED DESCRIPTION OF THE INVENTION
A new family of organosulfur compounds of the general formula (2) is described herein,
Figure imgf000007_0001
(2) wherein Ri and R2 are linear or branched C1- C5 alkyl; linear or branched C1- C5 alkenyl, with the proviso that R1 is not prop-1-enyl (allyl); substituted linear or branched C1- C5 alkenyl; substituted linear or branched C1- C5 alkyl; in which the substituents are selected from OR3; NR4R5; COOR6;
CONR7R8; substituted or unsubstituted aromatic, in particular para-methoxybenzyl or orf/7O,para-methoxybenzyl; substituted or unsubstituted heteroaromatic, in which
R3 is selected from H, CORg, para-methoxybenzyl and trialkylsilyl, in which R9 is alkyl or substituted alkyl;
R4 and R5 are independently alkyl or R4 and R5 together form a phthalimido group; R6 is alkyl or substituted alkyl; and
R7 and R8 are independently alkyl or substituted alkyl.
More particularly, R1 is not allyl.
Preferred alkyl groups include methyl, ethyl, propyl, butyl, isopropyl and isobutyl. Preferred alkenyl groups include prop-1-enyl (allyl), 1-propenyl, 1-butenyl, 2- butenyl, 3-butenyl and 1-methyl-2-butenyl.
For example, R2 can be prop-1-enyl and R1 can be any one of propyl, 3- hydroxypropyl, 3-phthalimidopropyl, te/t-butyl, benzyl, para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl, 3-(ferf-dimethylsilyloxy)propyl; or R1 can be para- methoxybenzyl and R2 can be one of para-methoxybenzyl, benzyl, para-fluorobenzyl; or R-i can be dansyl and R2 can be propyl. More specific examples of compounds of formula (2) are shown in Table 1. Table 1: Examples of compounds of formula (2)
Figure imgf000009_0001
Figure imgf000010_0002
The compounds of the formula (2) can be used for inhibiting the growth of tumour cells. They can therefore be used for treating cancer, such as lung cancer, oesophageal cancer or breast cancer, by administering an effective amount of the compound to a patient in need of treatment. The compound would typically be included in a pharmaceutical composition with a pharmaceutically acceptable carrier. The composition may include a mixture of the E- and Z-isomers of the compound, only the E-isomer or only the Z-isomer.
The compounds can be made by the following method:
R1
(16)
acylating a compound of formula (16), wherein R1 is as described above, with thiolacetic acid to form a thioacetate compound of formula (17);
Figure imgf000010_0001
(17)
generating a thiol of (17) by treatment of (17) with a base and then reacting the thiol with a compound of formula (18), prepared from a tosylate, halide or amide of a desired R2-substituent, to produce a compound of formula (19); and
Figure imgf000011_0001
oxidizing the compound of formula (19) to produce the compound of formula (2).
The compound of formula (16) can be prepared by reacting a substituted leaving group, such as a substituted halide of formula (25) (where X is a halide), with thiourea to form a corresponding thiourea salt (25a) and, typically in a one-pot reaction, reacting the salt with a base and with a propargyl halide or tosylate to form a compound of formula (26).
Figure imgf000011_0002
(25) (25a) (26)
The substituted halide of formula (25) is generally reacted with the thiourea in an aprotic solvent, such as acetonitrile. The thiouronium salt (25a) is generally hydrolysed in an alcoholic solvent such as methanol or ethanol using an alkali metal base such as sodium or potassium hydroxide. Suitable propargyl halides include propargyl bromide, chloride or iodide.
The compound of formula (16) can be acylated with thiolacetic acid via a radical mechanism using a radical initiator, such as azobisisobutyronitrile (AIBN) or a substituted variant thereof, such as 1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN), to initiate the reaction or using palladium (0) coupling of a vinyl halide.
The thiol of (17) can be produced by hydrolysis of (17) in an alcoholic solvent, such as methanol or ethanol, using an alkali metal base, such as sodium or potassium hydroxide. The compound of formula (19) can be oxidized by reaction with an oxidizing agent, such as m-chloroperoxybenzoic acid (m-CPBA), peroxybenzoic acid or hydrogen peroxide, to produce the compound of formula (2). A mixture of the E- and Z-isomers of the compound of formula (2) can be produced by this method. The E- and Z-isomers of the compound of formula (2), in which R2 is allyl, are represented by formula (20.1) and formula (20.2), respectively.
The applicant has found that the formation of the Z-isomer is generally favoured over the E-isomer in a 2:1 ratio.
Figure imgf000012_0001
E-isomer (20.1 ) Z-isomer (20.2)
The E- and Z-isomers may, in certain cases, be in the form of an inseparable mixture and, in other cases, in the form of a separable mixture. In the cases where the isomers are in the form of a separable mixture, they may be separated. Suitable separation methods include silica gel column chromatography; preparative TLC and high performance liquid chromatography (HPLC).
Example
The invention will now be described in more detail by way of the following non- limiting example.
The preparation of (EZZ)- 4,5,9-trithiadodeca-1 ,6-diene-9-oxide (3) is described below. It will, however, be apparent to a person skilled in the art that other compounds of formula (2) can readily be made using the same method.
1. toluene / AIBN
2. thiolacetic acid
Figure imgf000013_0002
Figure imgf000013_0003
Figure imgf000013_0001
Scheme 1 : Synthetic route to synthesize analogue (3)
In the above synthesis, 4-thia-1-heptyne (21) is converted to the thioester [E1Z)-
S-1-(4-hiahept-1-enyl) ethanethioate (22) with thiolacetic acid via radical addition using
1 ,1'-azobis(cyclohexanecarbonitrile) (ACCN) in toluene and as a 2:1 E:Z mixture of the thioester products (22). The thiol anion of (22) is then formed in KOH / methanol at - 4O0C (low temperature bath with acetonitrile / N2) and then alkylated using the allylated thiotosylate (24) to produce the disulfide 4,5,9-trithiadodeca-1 ,6-diene (23) in high yield. The sulfide is then oxidized up to the sulfoxide at low temperature with m- chloroperbenzoic acid in dichloromethane to give analogue (3) in 51% overall yield for
3 steps. The E- and Z- isomers were separated using silica gel column chromatography.
The detailed stepwise reaction sequence to produce compound (3) is shown below. All other analogues are synthesized according to the same procedure.
S-1-(4-Thiahept-1-enyl) ethanethioate (22)
Figure imgf000013_0004
Thiolacetic acid (7.35 g, 96.6 mmol), alkyne 21 (10.0 g, 87.6 mmol) and 1 ,1'- azobis(cyclohexanecarbonitrile) (ACCN) (2.14 g, 8.76 mmol) were refluxed in toluene at 80°C under N2 for 2 h. Toluene was then removed under vacuum and the residue dried on a pump to give an orange oil, which was purified directly on a silica-gel column using neat petroleum ether as eluent to give compound 22 (9.80 g, 80 % based on recovery of 2.64 g of alkyne 21 and as a 3:2 mixture of TJE stereoisomers: vmax / cm"1 2957 (C-H), 1698 (C=O), 634 (C-S): Z-22 δH (400 MHz, CDCI3) 0.95 (3H, t, J 7.4 Hz, H-10), 1.60 (2H, m, H-9), 2.37 (3H, s, H-1), 2.41 (2H, t, J 7.3 Hz, H-8), 3.16 (2H, dd, J 1.3, 7.8 Hz, H-6), 5.85 (1 H, dt, J 7.8, 10.4 Hz, H-5), 6.65 (1 H, dd, J 1.3, 10.4 Hz, H-4); δc (100 MHz, CDCI3), 13.4 (C-10), 22.8 (C-9), 30.8 (C-1), 31.0 (C-6), 33.2 (C-8) , 119.1 (C-4), 130.5 (C-5), 191.2 (C-9). £-22 δH (400 MHz, CDCI3), 0.96 (3H, t, J 7.4 Hz, H-10), 1.60 (2H, m, H-9), 2.32 (3H, s, H-1), 2.41 (2H, t, J 7.3 Hz, H-8), 3.21 (2H, dd, J 1.3, 7.8 Hz, H-6), 5.80 (1H, dt, J 7.8, 15.7 Hz, H-5), 6.52 (1 H, dt, J 1.3, 15.7 Hz, H-4); δc (100 MHz, CDCI3), 13.4 (C-10), 22.6 (C-9), 30.5 (C-1), 33.0 (C-8), 34.0 (C-6), 118.8 (C-4), 130.5 (C-5), 192.8 (C-9).
(E,Z)-4,5,9-trithiadodeca-1 ,6-diene (23)
Figure imgf000014_0001
KOH (1.44 g, 25.2 mmol) was dissolved in methanol (20 ml) at O0C and added dropwise to thioate ester 22 (2.40 g, 12.6 mmol) in methanol (10 ml) at -30°C under N2. The reaction was left stirring for 20 min before cooling to -78°C, whereupon S-prop-2- enyl 4-methylbenzenesulfonothioate (2.94 g, 12.9 mmol in methanol (5.0 ml) was added dropwise. The reaction was allowed to warm to O0C and left stirring for 2 h before quenching with aqueous ammonium chloride (20 ml). Water (40 ml) was added and the product was then extracted with CH2CI2 (3 x 50 ml). The CH2CI2 extracts were washed with saturated brine solution (2 x 20 ml), dried and reduced under vacuum. The resultant orange liquid was purified on a silica-gel column using petroleum ether as eluent to give compound 23 as a 3:2 mixture of Z:E geometrical stereoisomers (2.46 g, 89% yield):
Z- 23 δH (400 MHz, CDCI3) 0.99 (3H, t, J 7.3 Hz, H-12), 1.63 (2H, m, H-11), 2.48 (2H, t, J 7.3 Hz, H-10), 3.27 (2H, dd, J 1.1, 7.4 Hz, H-8), 3.37 (2H, m, H-3), 5.18 (2H, m, H-1), 5.69 (1H, dt, J 7.4, 9.7 Hz, H-7), 5.87 (1H, m, H-2), 6.23 (1H, dt, J 1.1, 9.7 Hz, H-6); δc (100 MHz1 CDCI3), 13.5 (C-12), 23.0 (C-11), 29.5 (C-8), 33.4 (C-10), 42.1 (C-3), 118.9 (C-1), 128.7 (C-7), 131.7 (C-6), 132.9 (C-2).
E-23 δH (400 MHz1 CDCI3), 0.99 (3H, t, J 7.3 Hz, H-12), 1.60 (2H, m, H-11), 2.45 (2H, t, J 7.3 Hz, H-10), 3.19 (2H, dd, J 1.2, 7.7 Hz, H-8), 3.35 (2H, m, H-3), 5.17 (1 H, d, J 8.9 Hz, H-1 trans), 5.29 (1H, d, J 16.8 Hz, H-1 cis), 5.86 (2H, m, H-2, H-7), 6.11(1H, dt, J 1.2, 14.6 Hz, H-6); δc (100 MHz, CDCI3), 13.5 (C-12), 22.7 (C-11), 33.1 (C-10), 33.5 (C- 8), 41.3 (C-3), 118.9 (C-1), 127.5 (C-7), 128.6 (C-6), 132.8 (C-2).
(E1Z)- 4,5,9-Trithiadodeca-1 ,6-diene 9-oxide (3)
Figure imgf000015_0001
3a 3b
m-Cpba (396 mg, 2.30 mmol) was dissolved in CH2CI2 (5 ml) and added dropwise to compound 23 (460 mg, 2.09 mmol) in CH2CI2 (5 ml) at -78°C under N2. The reaction was allowed to warm to room temperature and left stirring for 3 h, before quenching with saturated aq. sodium bicarbonate (15 ml) and extracting with ethyl acetate (3 x 30 ml). The organic extracts were dried with MgSO4, and the solvent removed under vacuum. The resultant residue was then purified on a silica-gel column using 40% ethyl acetate in petroleum ether as eluent to give compound 3 (400 mg, 82%) as a separable 3:2 mixture of Z:E geometrical stereoisomers.
Z-3a IR vmax (neat) / cm"1 2976 (HC=C), 2246 (HC-C aliphatic), 1368 (C=C), 1302 (S=O), 651 (C-S), 450 (S-S); δH (400 MHz, CDCI3) 1.08 (3H, t, J 7.3 Hz, H-12), 1.81 (2H, m, H-11 ), 2.61 (1 H, dt, J 8.2, 13.7 Hz, H-1 Oa), 2.71 (1 H, dt, J 8.2, 13.7 Hz, H-1 Ob), 3.37 (2H, m, H-3), 3.56 (1 H, dd, J 8.1 , 13.2, Hz, H-8a), 3.63 (1 H, dd, J 8.1 , 13.2, Hz, H- 8b), 5.16 (2H, m, H-1), 5.76 (1 H, dt, J 8.1 , 9.2, Hz, H-7), 5.84 (1H, m, H-2), 6.54 (1H, d, J 9.2 Hz, H-6); δc (100 MHz, CDCI3), 13.4 (C-12), 16.2 (C-11), 42.1 (C-3), 50.9 (C-8), 53.5 (C-10), 118.5 (C-7), 119.2 (C-1), 132.6 (C-2), 138.2 (C-6). E-Zb IR vmax (neat) / cπf1 2970( HC=C), 2232 (HC-C aliphatic), 1400 (C=C), 1018 (S=O), 644 (C-S), (S-S). δH (400 MHz, CDCI3), 1.06 (3H, t, J 7.3 Hz, H-12), 1.80 (2H, m, H-11), 2.61 (2H, m, H-10), 3.33 (2H, m, H-3), 3.43 (1 H, dd, J 7.9, 13.2 Hz, H-8a), 3.51 (1H, dd, J 7.9, 13.2 Hz, H-8b), 5.15 (2H, m, H-1), 5.82 (1H, m, H-2), 5.91 (1H, dt, J 7.9, 14.6 Hz, H-7), 6.34 (1H, d, J 14.6, H-6); δc (100 MHz, CDCI3), 13.3 (C-12), 16.1 (C-11), 41.2 (C-3), 53.0 (C-10), 54.5 (C-8), 117.1 (C-7), 119.1 (C-1), 132.4 (C-2), 134.2 (C-6).
Other compounds synthesized by this synthetic route include: (E/Z)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (3), or [EZZ)- 4,8,9-trithiadodeca- 6,11-diene-1-ol-4-oxide (4), (£/Z)-12-phthalimido-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (5), (£/Z)-10,10-dimethyl-4,5,9-trithiaundeca-1,6-diene-9-oxide (6), EZZ)-10-phenyl- 4,5,9-trithiadeca-1,6-diene-9-oxide (7), (EZZ)- 10-(p-methoxyphenyl)-4,5,9-trithiadeca- 1 ,6-diene-9-oxide (8), [EZZ)A 2-(p-methoxybenzyloxy)-4,5,9-trithiadodeca-1 ,6-diene-9- oxide (9), (£/Z)-1-(p-fluorophenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7- oxide (10), (EZZ)-I -(p-methoxyphenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7- oxide (11), (E/Z)-1-phenyl-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide (12), (E/Z)-1-(dansylamino)-4,5,9-trithiadodeca-6-ene-9-oxide (13), (EZZ)- 4,5,9- trithiadodeca-6-ene-9-oxide (14) and (EZZ)- 2, 3, 7-trithiadeca-4-ene-7-oxide (15).
in vitro anti-cancer activity
The compounds were evaluated for their in vitro ability to inhibit cell growth of cultured tumour cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. These cell types include:
1. WI38 cells which are normal embryonic lung fibroblasts; 2. CT-1 cells which are transformed WI38 fibroblasts;
3. WHC01 cells which are oesophageal epithelial cancer cells; and
4. EPC2 cells which are normal oesophageal epithelial cells; and
5. MDA-MB-231 cells which are human breast epithelial cancer cells; and
6. MCF12a cells which are normal human breast epithelial cells.
One oesophageal squamous cell carcinoma cell line WHCO1 originally established from surgical biopsies of primary oesophageal squamous cell carcinomas, a transformed fibroblast cell line CT-1 or human breast epithelial cells MDA-MB-231 were cultured in DMEM containing 10% foetal calf serum and 1% penicillin and streptomycin at 370C in a humidified atmosphere of 5% CO2. For the MTT assay, 3 x 103 cells were plated in 96-well plates in 90 μL DMEM per well. Compounds solubilized in DMEM + 1% DMSO (10 μl) were added to cells and DMEM + 1% DMSO, (10 μl) alone was added to the control to give a final concentration of 0 - 200 μM compound and 0.1% DMSO. After 48 hours incubation period, 10 μL of the MTT labelling reagent (final concentration 0.45 mg / ml) was added to each well and incubated for 4 hours in a humidified atmosphere at 37°C. One hundred micro litres of the solubilization solution was added to each well and the plates were incubated overnight at 370C. The spectrophotometric absorbance of the wells was measured at 595 nm using a microtiter plate reader. 0
The results are displayed as in vitro IC50 1S in Table 2, defined as the concentration of drug required to inhibit cell growth of half of the cell population.
5 Table 2: In vitro ICgfΛ/alues Obtained for Aioene-Analogues on CT-1, WHC01 and 1 MDA cell lines
CT1 WHC01 MDA
Compound
IC50/ IC50/ IC50/ number n 95 % Cl n 95 % Cl n 95 % Cl μM μM μM
E-ajoene 6 17.6 17.4-17.8 6 20.9 20.7-21.1 ND Z-ajoene 6 15.5 15.3-15.6 6 20.5 20.2 - 20.8 ND
E-3 5 26.7 26.5-26.9 10 35.8 35.5-36.0 2 25.9 25.5 -26.2 Z-3 5 17.0 16.8-17.2 12 21.6 21.5-21.7 5 19.2 19.0-19.3
E-4 5 23.1 23.0-23.3 6 26.8 24.7 - 29.2 ND Z-4 6 22.8 22.5-23.1 6 37.9 34.5-41.6 ND
E-5 4 95.9 95.0 - 96.8 5 68.9 61.2-77.6 ND Z-5 4 34.6 34.3 - 34.9 6 36.0 32.9 - 39.5 ND
EZ-6 5 33.1 32.5-33.6 6 25.4 23.4 - 27.6 ND
EZ-7 5 16.6 16.5-16.7 6 8.8 8.7-8.8 ND
EZ-8 12 11.2 11.1 -11.3 6 7.12 7.07-7.17 3 6.60 5.85-7.35 E-9 5 23.5 23.2 - 23.8 8 19.1 18.9 -19.3 ND Z-9 6 21.7 21.6 - 21.9 6 16.9 16.7 - 17.0 ND
EZ-10 ND 3 13.6 13.5 -13.7 ND
EZ-11 ND 2 1.34 1.33 - 1.36 4 0.82 0.82 - 0.83
EZ-12 ND 2 3.05 3.01 - 3.10 1 1.19
EZ-13 5 17.7 17.5 - 17.9 3 14.2 14.1 - 14.3 42.6
E-14 4 9.32 9.11 - 9.36 4 33.8 34.2 - 33.4 ND Z-14 ND ND ND
E-15 6 10.3 10.2 - 10.4 5 47.4 47.1 - 47.7 ND Z-15 ND ND ND
ND = not determined n = number on independent experimental determinations
IC50 = Inhibitory concentration of drug used to cause 50% inhibition of cell growth
95% Cl = 95% confidence interval
From the in vitro data, it was found that both the Z- and E- isomers of ajoene have equivalent activity at inhibiting cell growth of CT-1 and WHC01 cancer cells. For the synthesized ajoene analogues, with the exception of analogue 4 on WHCO1 cells, the Z-isomers are all more active at inhibiting cancer cell growth than their corresponding E-isomers.
Some of the analogues synthesized display superior activity to ajoene. In particular, the most active analogue EZ-11, is fifteen times more active than ajoene at inhibiting growth of WHCO1 oesophageal cancer cells in vitro. The four most active drug candidates that display IC50 activities under 10 μM are shown below.
Figure imgf000019_0001
(EZ-7) IC50 (WHCO1) = 8.8 uM
Figure imgf000019_0003
Figure imgf000019_0002
(EZ-8) IC50 (WHCO1) = 7.1 uM (EZ-11 ) IC50 (WHCO1 ) = 1.34 uM
It appears that strongly electron donating groups and / or lipophilic groups at Rio> Rii, Ri2 and Ri3 positions in the compound of formula (27) are important for strong in vitro activity. It also appears that lipophilic substituents at R1 and R2 in formula (2) are also important for good in vitro anti-cancer activity.
Figure imgf000019_0004
Formula (27): General structure of the active-class of compounds with R-10-R13 being electron donating / lipophilic groups.
In summary, the applicant has developed a synthetic route to access compounds of formula (2), being analogues of ajoene, with varying substituents at the Ri and R2 positions. The strongest drug candidates synthesized to date are those compounds with either benzyl or para-methoxybenzyl groups at positions R1 and/or R2 of formula (2). These compounds display in vitro IC50 killing activities on WHC01 and MDA cancer cells at drug concentrations under 10 μM.

Claims

CLAIMS:
1. A compound of formula (2)
Figure imgf000020_0001
(2) wherein:
R1 and R2 are linear or branched C1- C5 alkyl; linear or branched C1- C5 alkenyl, with the proviso that R1 is not prop- 1-enyl (allyl); substituted linear or branched C1- C5 alkenyl; or substituted linear or branched C1- C5 alkyl; in which the substituents are selected from OR3; NR4R5; COOR6; CONR7R8; substituted or unsubstituted aromatic; substituted or unsubstituted heteroaromatic, in which
R3 is selected from H, COR9, para-methoxybenzyl, and trialkylsilyl, in which R9 is alkyl or substituted alkyl;
R4 and R5 are independently alkyl or R4 and R5 together form a phthalimido group;
R6 is alkyl or substituted alkyl;
R7 and R8 are independently alkyl or substituted alkyl.
2. A compound according to claim 1 , wherein the alkyl groups are independently selected from the group consisting of methyl, ethyl, propyl, butyl, isopropyl and isobutyl.
3. A compound according to claim 1 or 2, wherein the alkenyl groups are independently selected from the group consisting of prop-1-enyl (allyl), 1- propenyl, 1-butenyl, 2-butenyl, 3-butenyl and 1-methyl-2-butenyl.
4. A compound according to any one of claims 1 to 3, wherein R1 is selected from the group consisting of propyl, 3-hydroxypropyl, 3-phthalimidopropyl, terf-butyl, benzyl, para-methoxybenzyl, ort/?o,para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl, dansyl and 3-(ferf-dimethylsilyloxy)propyl.
5. A compound according to any one of claims 1 to 4, wherein R2 is selected from the group consisting of propyl, prop-1-enyl, para-methoxybenzyl, ortho.para- methoxybenzyl, benzyl and para-fluorobenzyl.
6. A compound according to any one of claims 1 to 5, wherein R1 is selected from the group consisting of propyl, 3-hydroxypropyl, 3-phthalimidopropyl, ferf-butyl, benzyl, para-methoxybenzyl, o/#}o,para-methoxybenzyl, 3-(para- methoxybenzyloxy)propyl and 3-(tert-dimethylsilyloxy)propyl and R2 is prop-1- enyl.
7. A compound according to any one of claims 1 to 5, wherein R1 is para- methoxybenzyl or orf/?o,para-methoxybenzyl and R2 is selected from the group consisting of para-methoxybenzyl, orf/?o,para-methoxybenzyl, benzyl and para- fluorobenzyl.
8. A compound according to any one of claims 1 to 5, wherein R1 is dansyl and R2 is propyl.
9. A compound according to any one of claims 1 to 5, which is: (E/Z)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (3),
(EZZ)- 4,8,9-trithiadodeca-6,11-diene-1-ol-4-oxide (4),
(EZZ)A 2-phthalimido-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (5),
(E/Z)-10,10-dimethyl-4,5,9-trithiaundeca-1 ,6-diene-9-oxide (6),
(EfZ)A 0-phenyl-4,5,9-trithiadeca-1 ,6-diene-9-oxide (7),
(EZZ)- 10-(p-methoxyphenyl)-4,5,9-trithiadeca-1 ,6-diene-9-oxide (8),
(EZZ)A 2-(p-methoxybenzyloxy)-4,5,9-trithiadodeca-1 ,6-diene-9-oxide (9), (E/Z)-1 -(p-fluorophenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide
(10),
(£/Z)-1 -(p-methoxyphenyl)-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide
(11 ).
(E/Z)-1-phenyl-8-(p-methoxyphenyl)-2, 3, 7-trithiaocta-4-ene-7-oxide (12),
[EZZ)A -(dansylamino)-4,5,9-trithiadodeca-6-ene-9-oxide (13),
(EZZ)- 4,5,9-trithiadodeca-6-ene-9-oxide (14), or
(EZZ)- 2, 3, 7-trithiadeca-4-ene-7-oxide (15).
10. A compound according to any one of claims 1 to 9, for use in a method of killing or inhibiting the growth of tumour cells and/or treating cancer.
11. A compound according to claim 10, wherein the cancer is selected from the group consisting of lung cancer, oesophageal cancer, cervical cancer and breast cancer.
12. A pharmaceutical composition comprising a compound of any one of claims 1 to 10 and a pharmaceutically acceptable carrier.
13. A pharmaceutical composition according to claim 11 , for use in a method of killing or inhibiting the growth of tumour cells and/or for treating cancer.
14. A pharmaceutical composition according to claim 13, wherein the cancer is selected from the group consisting of lung cancer, oesophageal cancer, cervical cancer and breast cancer.
15. A method of making a compound of formula (2) according to any one of claims 1 to 11 , the method including the steps of:
(v) acylating a compound of formula (16), wherein R1 is as described in claim 1 , with thiolacetic acid to form a thioacetate compound of formula (17)
Figure imgf000022_0001
(16)
Figure imgf000023_0001
(17)
(vi) generating a thiol by treating the compound of formula (17) with a base; (vii) reacting the thiol with a compound of formula (18), prepared from a tosylate, halide or amide of R2, wherein R2 is as described in claim 1 , to produce a compound of formula (19)
Figure imgf000023_0002
(18) (19)
(viii) and oxidizing the compound of formula (19) to produce the compound of formula (2)
S S R2
H O
(2)
16. A method according to claim 15, wherein the compound of formula (16) is acylated with thiolacetic acid via a radical mechanism using a radical initiator to initiate the reaction.
17. A method according to claim 16, wherein the radical initiator is azobisisobutyronitrile (AIBN) or a substituted variant thereof.
18. A method according to claim 17, wherein the substituted variant is 1 ,1 '- azobis(cyclohexanecarbonitrile) (ACCN).
19. A method according to claim 15, wherein the compound of formula (16) is acylated with thiolacetic acid using palladium (0) coupling of a vinyl halide.
20. A method according to any one of claims 15 to 19, wherein the thiol is produced by hydrolysis of the compound of formula (17) in an alcoholic solvent using an alkali metal base.
21. A method according to claim 20, wherein the alkali metal base is sodium or potassium hydroxide.
22. A method according to claim 20 or 21 , wherein the alcoholic solvent is methanol or ethanol.
23. A method according to any one of claims 15 to 22, wherein the compound of formula (19) is oxidized by reacting it with an oxidizing agent to produce the compound of formula (2).
24. A method according to claim 23, wherein the oxidizing agent is selected from the group consisting of m-chloroperoxybenzoic acid (m-CPBA), peroxybenzoic acid and hydrogen peroxide.
25. A method according to any one of claims 15 to 24, which further includes the step of:
(ix) separating the E- and Z-isomers of the compound of formula (2).
26. Use of a compound of any one of claims 1 to 11 in the manufacture of a medicament for the inhibition of the growth of tumour cells and/or for the treatment of cancer.
27. The use according to claim 26, wherein the cancer is selected from the group consisting of lung cancer, oesophageal cancer, cervical cancer and breast cancer.
28. A method of inhibiting the growth of tumour cells and/or treating cancer, the method including the step of administering to a person or animal in need of treatment a pharmaceutically effective amount of a compound of any one of claims 1 to 11.
29. A method according to claim 28, wherein the cancer is selected from the group consisting of lung cancer, oesophageal cancer, cervical cancer and breast cancer.
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CN102171184A (en) 2011-08-31
EP2321271A4 (en) 2012-07-25

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