GB2346325A

GB2346325A - Formulation comprising a brassica extract or sulforaphane and resveratrol

Info

Publication number: GB2346325A
Application number: GB9902304A
Authority: GB
Inventors: Raymond Sidney Matthews
Original assignee: WASSEN INT Ltd
Current assignee: WASSEN INT Ltd
Priority date: 1999-02-02
Filing date: 1999-02-02
Publication date: 2000-08-09
Also published as: WO2000045829A1; GB9902304D0; AU2305900A; GB2363571A8; GB2363571A; GB0118267D0

Abstract

The present invention discloses a composition suitable for pharmaceutical use which comprises at least one active ingredient from a brassica extract or an analogue of sulforaphane, and resveratrol or an analogue thereof. Suitably the composition comprises 1-100 mg of the brassica extract or sulforaphane analogue and 0.5-100 mg of resveratrol. Preferably the weight ratio of brassica extract to resveratrol is from 1:500 to 1:50. The composition may be used to treat cancer, especially testicular cancer.

Description

Formulation The present invention relates to compositions for retarding and/or preventing tumours.

There have been numerous attempts at treating tumours. It is accepted that tumour development can be a multistage process, ahd that it can be inhibited by interfering with various discrete elements in the overall process. Inhibition of the earliest stages would generally be considered the most desirable protective effect.

Many compounds have been tested for their efficacy in preventing tumours. One such compound is resveratrol. In vitro expeNiments have been undertaken with resveratrol and there is evidence from in vitro cell culture and bacterial mutagenicity studies that resveratrol might retard tumour initiation when a mutagenic or carcinogenic challenge is given. The improved scavenging of free radicals, possibly by the induction of the phase II enzyme quinone reductase, has been proposed to account for the protection of the cells (Jang t al, Science, 275,218-20 (1997)).

There is also evidence that resveratrol might inhibit cyclooxygenase, an enzyme considered to be involved in tumour promotion and various inflammatory conditions.

Anti-proliferative effects of resveratrol have, been noted in vitro and attributed to an inhibition of thmidine incorporation and the inhibition of esterase enzymes (Jang et al., (1997)). In addition, in a leukaemia cell vine, a marked inhibition of the enzyme ribonucleotide reductase has been reported (Tontcave et al., FEBS Letters, 421, 277-9 (1998)) and this should also have an anti-prqliferative effect in vivo.

Resveratrol is a stilbene. It has two forms, the trans form and the cis form. A limited number of stilbene-containing plants have bqen consumed by man, and of these, the best known is the grape.

Several organic isothiocyanates have been tested for anti-cancer activity. One of these is sulforaphane which can be extracted from plants of the genus Brassica. It has been shown that sulforaphane does not induce phase I drug metabolising enzymes ( Zhang et al (1992)). Later studies have shown that sulforaphane increased the activity of 2 isoforms of glutathione-S-transferase and decreased the major human cytochrome P450 CYP3A4 (Mahoe et al., Cancer Res. 57,3649-3652 (1998)).

Sulforaphane has been reported to reduce the activation of the Aflatoxin B, by human hepatocytes (Longuet et al. Molecular Toxicology 11,95-191,1998) and to reduce the incidence and multiplicity of mammary tumours following administration of DMBA (Verhoeven et al., 1997).

Some in vivo studies have also been performed on sulforaphane. Two studies showed a reduced binding of the aflatoxin B, following the administration of diets of cabbage and brussel sprouts. Reductions of DMBA or MNU-induced mammary tumours in rats have been found in 3 studies when cabbage, cauliflower, broccoli or brussel sprouts were included in the diet. Administration of cauliflower reduced the liver carcinogenesis induced by AFB I. A similar study, also with AFB 1 showed a reduction in the number of tumours in the liver while a study on mice receiving cabbage along with DMH showed a reduction in the total number of tumours.

Sulforaphane is one of a number of organic thiocyanates released on hydrolysis of the aliphatic glucosinolates.

The present invention is based on the fact khat a combination of resveratrol or its analogue and a brassica extract is surprisingly effective in treating tumours, especially testicular tumours. Accordingly, the present invention provides a composition suitable for pharmaceutical use which comprises at least one active ingredient obtainable from a brassica extract or an analogue of sulforaphane and resveratrol or an analogue thereof.

Resveratrol can also be obtained from plants. The composition can thus be obtained by mixing the plant extracts. The brassica extract may be obtained from any brassica vegetable which includes cabbage, kale, cauliflower, brocoli, mustard greens, kohlrabi, brussels sprouts and horseradish. The brassica extract is preferably a brocoli extract. Resveratrol may be extracted from grapes or other parts of grapevines or made via a synthetic preparation.

The analogues of resveratrol include stilbenfs, hydroxylated stilbenes, for example trihydroxy-stilbenes and tetrahydroxy-stilbepes, which are typically phytoalexins, with or without one or more attached sugars or alkyl groups such as methyl; oligomers and/or polymers thereof, as well as oxidation or reduction products thereof. In particular, 3,4', 5-trihydroxystilbete-3-beta-mono-D-glucoside or resveratrol which is preferred, may be used and the pharmacologically acceptable salts and esters thereof.

The sulforaphane analogue which can be used in the composition may be any compound having an isothiocyanate moiety tnd a polar functional group moiety, wherein the two moieties are linked by a chain of one or more carbon atoms and the compound contains no pyridyl moieties, or a pharmacologically acceptable salt of such a compound.

The sulforaphane analogue is preferably not a heteroaromatic compound and is preferably not an arylalkyl compound. The analogue is preferably an olefin and is preferably aliphatic. The second moiety is preferably a polar functional group selected from a carboxylic ester, a carboxylic acid, a hydrocarbonoxy, a halogen, a hydroxyl, a ketone, a cyano, a nitro, a phosphine oxide, a sulfide, a sulfone, a sulfoxide, a thioether, and a thioester group, more preferably selected from a hydroxyl, a ketone, a phosphine oxide, a sulfone, and a sulfoxide group. The carbon chain of the sulforaphane component preferably comprises at least 3 carbon atoms, more preferably 3 to 5 carbon atoms. The carbon chain is preferably part of a non aromatic ring.

The sulforaphane component is preferably selected from sulphoraphane itself, sulforaphene (4-isothiocyanato-(lR)-(methylsulfinyl)-l-(E)-butene), 6 isothiocyanato-2-hexanone, exo-2-acetyl-6-isothiocyanatonorbornane, exo-2- isothiocyanato-6-methylsulfonylnorbornane, 6-isothiocyanato-2-hexanol, 1- isothiocyanato-4-dimethylphosphonylbutane, exo-2- (1'-hydroxyethyl)-5- isothiocyanatonorbornane, exo-2-acetyl-5-isothiocyanatonorbornane, 1- isothiocyanato-5-methylsulfonylpentane and cis-or trans-3- (methylsulfonyl) cyclohexylmethylisothiocyanate and is preferably either form of sulforaphane, more preferably ( (-) 1-isothiocyanato- (4R)- (methylsulfinyl) butane).,-, Bertoin, alyssin, erucin, erysolin, iberverin, iberin and cheirolin may also be used.

Although the brassica extract and resveratrol and its analogues both show potential for reducing the incidence of cancers, surprisingly, when used together, they show a synergistic effect. The brassica extract or analogue of sulforaphane appears to act principally on the initiation phase of carcinogenesis. Whereas, resveratrol and its analogues may inhibit protein kinases in vivo and therefore affects the subsequent proliferative phase of cancer.

A further aspect of the invention provides for the use of the composition of the invention for treating tumours. The composition can be used in a method of treatment of In a further aspect, the composition may additionally comprise pharmaceutically acceptable diluents or excipients. It may alo comprise antioxidant compounds, vitamins and minerals, in particular, vitamip A, vitamin C, vitamin E, lycopene and selenium.

The composition preferably comprises the active ingredient obtainable from a brassica extract or a sulforaphane analogue nd resveratrol or its analogue in a weight ratio of 1 : 1000 to 1 : 10, preferably 1 : 500to i : 50, more preferably 1: 150 to 1: 75 and especially about 1 : 100. This last formulatioU typically contains sulforaphane and resveratrol in a ratio of 2 : 1.

The composition is preferably administered in doses containing I to 100mg of the active ingredient obtainable from a brassica extract or sulforaphane analogue and 0.5 to l OOmg of resveratrol or an analogue, preferably in a dose of l Omcg of active ingredient obtainable from a brassica extract or sulforaphane analogue: Img resveratrol or its analogues. The composition may be administered with a frequency of several times a day to once every two dayi, preferably daily. Treatment should be ongoing.!

Claims

CLAIMS 1. A composition suitable for pharmaceutical use which comprises at least one active ingredient from a brassica extract or an analogue of sulforaphane, and resveratrol or an analogue thereof.
2. A composition according to claim I in which the resveratrol or analogue thereof is an extract of a grape or a grapevine
3. A composition according to claim 1 or 2 which comprises resveratrol
4. A composition according to any one of the preceding claims which comprises sulforaphane.
5. A composition according to any one of the preceding claims which comprises brassica extract and in which the weight ratio of brassica extract to resveratrol or its analogue is from 1: 500 to 1: 50.
6. A composition according to any one of the preceding claims which comprises 1 to l OOmg of the brassica extract or sulforaphane analogue and 0.5 to I OOmg of resveratrol or analogue thereof.
7. A composition according to any one of the preceding claims which further comprises one or more of vitamin A, vitamin C, vitamin E, lycopene, lipoic acid, limonene, selenium and bromelain.
8. A composition according to claim 1 substantially as hereinbefore described.
9. A process for producing a composition according to any one of the preceding claims which comprises mixing the active ingredients together.
10. A composition according to any one of claims 1 to 8 for use in a method of treatment of the human or animal body by therapy.
11. Use of a composition according to any one of claims I to 8 in the manufacture of a medicament for use in the treatment of tumours.
12. Use according to claim 11 of a composition according to any one of claims I to 8 in the treatment of testicular tumours.
13. A product containing at least one active ingredient from a brassica extract or an analogue of sulforaphane, and resveratrol or an analogue thereof, for simultaneous, separate or sequential use in the treatment of tumours.