AU2008200718B2 - Mevalonic acid derivatives - Google Patents

Description

AUSTRALIA Patents Act COMPLETE SPECIFICATION (ORIGINAL) Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: Ohgen Research Laboratories Ltd. Actual Inventor(s): Kiyoshi Akiyama Address for Service and Correspondence: PHILLIPS ORMONDE & FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: MEVALONIC ACID DERIVATIVES Our Ref: 822663 POF Code: 1349/160761 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1 - 2 5 10 SPECIFICATION 15 MEVALONIC ACID DERIVATIVES Technical Field This invention relates to novel mevalonic acid derivatives and to 20 their utilization as antitumor agents. Background Art Many geranyl compounds having 1,5-diene structure are present in vivo, and are known as in vivo precursors of substances having polyene 25 structure and exhibiting various physiological activities. These substances having 1,5-diene structure and polyenes derived therefrom invariably start from mevalonic acid and biosynthesized. I noticed, as such geranyl compounds having 1,5-diene structure, geranic acid or geranylamine, and furthermore mevalonic acid which is the 30 base for biosynthesis of polyenes, synthesized various derivatives of geranic acid or geranylamine and mevalonic acid and investigated their physiological activities, in particular, antitumor activity and toxicity, and have come to complete the present invention.

3 The discussion of the background to the invention herein is included to explain the context of the invention. This is not to be taken as an admission that any of the material referred to was published, known or part of the common general knowledge in Australia as at the priority date of any 5 of the claims. Throughout the description and claims of the specification the word "comprise" and variations of the word, such as "comprising" and comprises", is not intended to exclude other additives, components, integers or steps. 10 Disclosure of the Invention This invention provides mevalonic acid derivatives represented by the following formula (I-4): R4 15 HO O H H OH H

CH

3 (1-4) H OH | H NH-C-CH 2 -- ;-C-CH 2

CH

2 -OH 20 0 OH in which R 4 stands for -CH 2 OH or -CH 3 . The mevalonic acid derivatives of above formula (1-4) include, for example, the following: 25 N-glucosylmevalonamide

CH

2 OH H O H H OH H (25) CH 30 HO OH CH, H NH-C-CH -C-CH 2

CH

2 OH 0 OH N-galactosylmevalonamide 4 CH 20H HO O H H OH H HCH (26) HOH OH3 5 H NH-C--- I (CH2CH2OH O OH and N-fucosemevalonamide 10

CH

3 HO O H H OH H H OH

H

3 (27) 15 H NH-C- CH2--CH2CH20H O OH Those mevalonic acid derivatives of the formula (1-4) can be prepared, for example, by reacting sugaramine represented by the following formula: 20 R4 HO O H H OH H (mi) H OH 25 H NH2 in which R4 has the earlier given signification, or salt thereof with mevalolactone or mevaloyl halide. Said reaction of a sugaramine of the formula (III) or a salt 30 thereof with mevalolactone or mevaloyl halide (e.g., mevaloyl chloride) can be conducted in water or an adequate inert organic solvent (e.g., N,N-dimethylformamide, tetrahydrofuran, chloroform or 5 the like) at temperatures between room temperature to reflux temperature of the solvent, preferably from about 400 to about 70 0 C. The use ratio of mevalolactone or mevaloyl halide to a sugaramine of the formula (III) is not strictly limited, but it is 5 normally preferred to use 1-2 moles of mevalolactone or mevaloyl halide per mole of the sugaramine of the formula (III). Where a salt of a sugaramine of the formula (III) or mevaloyl halide is used as the starting material, it is generally desirable to carry out the above reaction with addition of a base, for example, 10 tertiary amine such as N-methylpiperidine; or an inorganic base such as sodium hydroxide, potassium hydroxide, potassium carbonate and the like. The mevalonic acid derivatives of the formula (1-4) produced through above reactions can be isolated from the reaction mixtures 15 and purified by conventional means, such as extraction, crystallization, chromatography or the like. Compounds of the formula (1-4) offered by the present invention possess excellent antitumor activity, as is clear from the following measurement results of antitumor effect. 20 Measurement of antitumor effect Carcinoma of HuH-7 cells (dendriform cell strain of human hepatoma) hypodermically implanted or subimplanted in the backs of 5-weeks old female nude mice (BALB/c, Ninox) was aseptically taken 25 out and crushed into 5 x 5 mm sized pieces in a phosphate buffer solution (PBS), a piece of which then being hypodermically implanted in the backs of the nude mice. Each test substance was dissolved in corn oil, and the solution was intraperitoneally administered to the nude mice consecutively 30 once per day at a rate of 250 jig/mouse for 3 weeks, starting from a week after the implantation. After termination of the administration, the carcinomas were taken out and weighed to calculate the antitumor effect and the mice' weight loss by the following equations. For the test six mice per group were used, and 35 the group administered with the solvent (corn oil) alone was made the 6 control group: Antitumor effect (%) average tumor weight of a test group 5 ~ x 100 average tumor weight of the control group Weight loss (%) average body weight of a test group mice 10 X 100 average body weight of the control group mice Evaluations of antitumor effect and weight loss were conducted according to the following standard, where the control 15 group values were held to be 100%. Antitumor effect -: >100%, +/-: 100-75%, +: 75-50%, ++: 50-25%, +++: 25-0% 20 Weight loss -: >110%, +/-: 110-100%, +: 100-95%, ++: 95-90%, +++: <90% Death rate (death rate during the test period) -- none 25 +/- death occurred with high concentration administration (500 pig/mouse) +: 1-3 mice dead .++: 3-5 mice dead +++: all 6 mice dead 30 Synthetic evaluation -: weak antitumor effect and very strong toxicity against host mice +/-: weak antitumor effect recognizable and toxicity against host 35 mice also observable +: a fixed level of antitumor effect observable but toxicity 7 against host mice also strong ++: strong antitumor effect observed and toxicity against host mice week +++: strong antitumor effect observed and no toxicity against 5 host mice The results are shown in the following Tables 1-3. Table 1 10 Test Substance Antitumor W ih Dth Synthetic Effect Weight Death Evaluation loss rate N-geranyl glucuronamide ++ +/- - ++ N-geranyl galacturonamide N-galactosylgeranamide ++ + -+++ N-fucosegeranamide ++ +- -+++ Table 2 'Test Substance Antitumor 'Ibxicity Synthetic Effect Weight Death Evaluation loss rate N,N'-digeranylmalic diamide + + - ++ N,N'-digeranylfumaric diamide ++ +/- - ++ N-geranyl-4 pyruvoamnnobenzamide - + N-geranoyltyrosine ++ +/- - ++ Tyrosine geranylamide + - + N-acetyltyrosine geranylamide + +/- - + 8 Table 3 rbst Substance Antitumor Toxicity Synthetic Effect Weight Death Evaluation loss rate N-glucosylmevalonamide ++ +- -+++ As is clear from the shown results, the compounds of the invention 5 of the formula (1-4) possess excellent antitumor effect against HuH-7 cells and furthermore almost no toxicity, and are expected to be useful as antitumor agents for treatment and therapy of various solid cancer represented by liver cancer. Where a compound of the present invention is used as a 10 medicine such as an antitumor agent, it can be administered orally or parenterally (e.g., intravenous injection, intramuscular injection, hypodermic injection, or the like). The effective dose is variable over a broad range depending on individual patients' symptoms, seriousness of the illness, body weight, age, and the doctor's diagnosis, 15 etc. Normally, however, taking a case of administration by injection, the dose can be about 1 - about 50 mg/kg/day, which may be administered at a single time or at plural times dividedly in a day. Where a compound of the present invention is used as a medicine, an effective dose of the compound can be formulated with 20 pharmaceutically acceptable carrier or diluent (e.g., excipient, solvent or other adjuvants) into a preparation form suitable for unit dose administration, for example, tablet, powder, granule, capsule, enteric coated pill, troche, syrup, elixer, liquid, suspension, emulsion and the like. 25 As carriers or diluents useful for the formulation, for example, excipients such as starch, lactose, sucrose, mannitol, carboxymethyl cellulose and the like; lubricants such as magnesium stearate, sodium laurylsulfate, tale and the like; binders such as dextrin, microcrystalline cellulose, polyvinylpyrrolidone, gum Arabic, corn 30 starch, gelatin and the like; disintegrators such as potato starch, carboxymethyl cellulose and the like; and diluent solvents such as 9 water for injection, physiological saline, aqueous dextrose solution, vegetable oil for injection, propylene glycol, polyethylene glycol and the like can be named. Furthermore, taste- or odor-correcting agent, colorant, isotonicity, stabilizer, antiseptic, soothing agent and the like 5 may be incorporated where necessary. In the pharmaceutical preparations according to the invention, moreover, other pharmacologically active substance(s) may be incorporated where necessary. Hereinafter the invention is explained still more specifically, 10 referring to working Examples. Examples Synthesis Example 1: Synthesis of N-glucosylmevalonamide 15 Glucosamine hydrochloride (2.16 g, 10 mmols) was dissolved in 20 ml of water, and to the aqueous solution 10 ml of 1N sodium hydroxide and mevalolactone (1.30 g, 10 mmols) were added, followed by 5 hours' heating under stirring at 55 0 C. After termination of the reaction, the reaction mixture was condensed under reduced pressure. 20 To the residue 100 ml of methanol was added and whereupon separated precipitate was filtered off. The filtrate was condensed again with an evaporator and the residue was separated on silica gel column chromatography, to provide 1.45 g of the object product from the ethanol distillate. The yield was 47%. Because the as-obtained 25 product was a viscous oily substance, a minor amount of dichloromethane was added thereto to effect crystallization. Upon suction filtering, 1.10 g of the title compound was obtained, which was strongly hygroscopic and its melting point could not be measured: 30 1 H NMR(DMSO-d6) 6=1.00(3H, s), 1.44-1.59(2H, m), 2.47(2H, s), 2.96-3.74(10H, m), 4.04-5.08(3H, m).

10 Synthesis ExamIe 2: Synthesis of N-galactosylmevalonamide Synthesis Example 1 was repeated except that the glucosamine hydrochloride was replaced with galactosamine hydrochloride, to provide the title compound: 5 IH NMR(DMSO-d6) 8=1.08(3H, s), 1.51:1.61(2H, m), 2.44(2H, s), 2.74-5.16(13H, m). Synthesis Example 3: Synthesis of N-fucosemevalonamide 10 Synthesis Example I was repeated except that the glucosamine hydrochloride was replaced with fucosamine hydrochloride, to provide the title compound: 'H NMR(DMSO-d6) 8=1.06(3H, s), 1.20(3H, d, J=24.OHz), 15 :1.54-1.62(2H, m), 2.44(2H, s), 2.74-5.15(12H, m). Formulation Example Two (2) g of N-glucosylmevalonamide was dissolved in 1 liter of water for injection at ambient temperature, isotonized with sodium 20 ~ chloride and sealed into ampoules. One (1) ml of this injection contains 2 mg of the active ingredient. 25 30

Claims (7)

1. Mevalonic acid derivatives represented by the following formula (I-4): R4 5 HO OH H OH H CH H OH 1 (1-4 H NH-C-CH 2 -- C--CH 2 CH--OH ii I 10 0 OH in which R 4 stands for -CH 2 OH or -CH.

2. An antitumor agent containing, as the active ingredient, a mevaloic acid derivative of the formula (1-4) as given in (aim 1. 15

3. A pharmaceutical composition comprising an activity-effective amount a mevalonic acid derivative of the formula (I-4) as given in Claim 1, and a pharmaceutically acceptable carrier or diluent. 20

4. Method of treating cancer comprising administration of antitumorically effective amount of a mevalonic acid derivative of the formula (1-4) as given in Claim 1 to a patient.

5. Use of a mevalonic acid derivative of the formula (1-4) as given in 25 Claim 1 for treatments against cancer.

6. Use of a mevalonic acid derivative of the formula (1-4) as given in Claim 1 for production of pharmaceutical preparations. 30

7. A mevalonic acid derivtive according to claim 1 substantially as herein described with reference to any one of the Examples.