[0001] The present invention relates to compositions for the treatment and/or amelioration of cancer. In particular, the compositions of the invention have an inhibitory effect on Lewis lung carcinoma in vivo, and have an inhibitory effect on other types of lung carcinoma, melanoma, prostate cancer, renal cancer, CNS cancer, colon cancer, ovarian cancer and breast carcinoma in vitro. The invention therefore also relates to the use of the compositions in methods of treating and/or ameliorating such tumours.

BACKGROUND ART

[0002] Recent interest in, and preference for, natural medicines and products has revitalized interest in plant oils. One such oil is terpinen-4-ol which has been widely studied by many researchers. Terpinen-4-ol is a naturally occurring monoterpene, and has been shown to have a number of biological activities, including antibacterial and antifungal.

[0003] A compound related to monoterpenes is the alkylbenzene, p-cymene. The inventor has found that when terpinen-4-ol and p-cymene are combined with p-menth-2-cn- 1 ,4-diol (1,4- dihydroxy-p-menth-2-ene) and p-menthanc- 1 ,2,4-triol. complete tumor apoptosis in a mice xenograft model is achieved. Therefore, whilst the biological functions of p-menth-2-en- 1 ,4-diol and / -menthane- 1 ,2,4-triol are not well known, they clearly play an important role when combined with the monoterpene, terpinen-4-ol, and p-cymene.

[0004] It will be clearly understood that, if a prior art publication is referred to herein, this reference does not constitute an admission that the publication forms part of the common general knowledge in the art in Australia or in any other country.

SUMMARY OF INVENTION

[0005] The present invention is directed to a composition comprising one or more monoterpenes in combination with / -mcnih-2-en- 1 ,4-diol and p-menthane- 1 ,2,4-triol which may at least partially overcome the difficulties of treating cancers, such as Lewis lung carcinoma, or provide the consumer with a useful or commercial choice.

[0006] With the foregoing in view, the present invention in one form, resides broadly in a composition comprising at least one monoterpene in combination with p-menth-2-en-l,4-diol. [0007] In a second broad form, the invention resides in a composition comprising at least one monoterpene in combination with p-menlhane- 1 ,2,4-triol.

[0008] In a third broad form, the invention resides in a composition comprising at least one monoterpene in combination with p-menth-2-cn- l ,4-diol and p-menthane- 1 ,2,4-triol.

[0009] In a fourth broad form, the invention resides in a composition comprising terpinen-4- ol, p-cymene, p-menth-2-en- l,4-diol and p-menthane- 1 ,2,4-triol.

[0010] In a fifth broad form, the invention resides in a method of preparing a composition comprising at least one monoterpene in combination with p-menth-2-cn- 1 ,4-diol.

[0011] In a sixth broad form, the invention resides in a method of preparing a composition comprising at least one monoterpene in combination with p-menthane- 1 2,4-triol.

[0012] In a seventh broad form, the invention resides in a method of preparing a composition comprising at least one monoterpene in combination with p-mcnlh-2-en- 1 4-diol and p-menthane-l,2,4-triol.

[0013] In an eighth broad form, the invention resides in a method of preparing a composition comprising terpinen-4-ol, p-cymene, p-menth-2-en- 1 ,4-diol and p-menthane- 1 ,2,4- triol.

[0014] In a ninth broad form, the invention resides in a method of treating cancer, comprising administering a composition comprising at least one monoterpene in combination with p-menth-2-en- l,4-diol.

[0015] In a tenth broad form, the invention resides in a method of treating cancer, comprising administering a composition comprising at least one monoterpene in combination with p-menthane- 1, 2, 4-triol.

[0016] In an eleventh broad form, the invention resides in a method of treating cancer, comprising administering a composition comprising at least one monoterpene in combination with p-menth-2-en- l,4-diol and p-menthane- 1,2, 4-triol.

[0017] In a twelfth broad form, the invention resides in a method of treating cancer, comprising administering a composition comprising terpinen-4-ol, p-cymene, p-menth-2-en-l,4- diol and p-menthane- 1,2, 4-triol.

[0018] In a thirteenth broad form, the invention resides in use of a composition comprising at least one monoterpene in combination with p-menth-2-cn- l ,4-diol in the manufacture of a medicament for treating cancer.

[0019] In a fourteenth broad form, the invention resides in use of a composition comprising at least one monoterpene in combination with p-menthane- 1 ,2,4-triol in the manufacture of a medicament for treating cancer.

[0020] In a fifteenth broad form, the invention resides in use of a composition comprising at least one monoterpene in combination with p-menth-2-en- 1 ,4-diol and p-menthane- 1 ,2.4-triol in the manufacture of a medicament for treating cancer.

[0021] In a sixteenth broad form, the invention resides in use of a composition comprising terpinen-4-ol, p-cymene, p-menth-2-en-l,4-diol and p-menthane- 1 ,2,4-triol in the manufacture of a medicament for treating cancer.

[0022] In a seventeenth broad form, the invention resides in a composition comprising at least one monoterpene in combination with p-menth-2-cn- l ,4-diol for treating cancer.

[0023] In an eighteenth broad form, the invention resides in a composition comprising at least one monoterpene in combination with p-menthane- 1 ,2,4-triol for treating cancer.

[0024] In a nineteenth broad form, the invention resides in a composition comprising at least one monoterpene in combination with p-menth-2-en- 1 ,4-diol and r-menthane- 1 ,2.4-triol for treating cancer.

[0025] In a twentieth broad form, the invention resides in a composition comprising terpinen-4-ol, p-cymene, p-menth-2-en-l,4-diol and p-menthane- 1 ,2,4-triol for treating cancer.

[0026] Monoterpenes are a class of compounds produced in a variety of plants. They consist of two isoprene units, have the molecular formula C ₁₀ H ₁₆ or C ₁₀ H ₁₈ O, and can be linear or cyclic. Examples of monoterpenes, including terpinen-4-ol, are illustrated in Table I, together with their building block, isoprene. The alkylbenzene, p-cymene, which is related to monoterpenes is also included. [0027] Table I - Examples of monoterpenes

[0028] To contrast, p-menth-2-cn- l ,4-diol has a molecular formula of C10H18O2, molecular weight of 170.248 g/mol, and the following structure:

and para-menthane- 1 ,2,4-triol has a molecular formula of C10H20O3, molecular weight of 188.26 g/mol, and the following structure:

[0029] Without wishing to be bound by theory, it is thought that combining a monoterpene, such as terpinen-4-ol, and p-cymene, with p-menth-2-cn- 1 ,4-diol and pura-menthane- l ,2.4-triol results in a conjugate compound.

[0030] Any of the features described herein can be combined in any combination with any one or more of the other features described herein within the scope of the invention.

[0031] The reference to any prior art in this specification is not, and should not be taken as an acknowledgement or any form of suggestion that the prior art forms part of the common general knowledge.

BRIEF DESCRIPTION OF DRAWINGS

[0032] Preferred features, embodiments and variations of the invention may be discerned from the following Detailed Description which provides sufficient information for those skilled in the art to perform the invention. The Detailed Description is not to be regarded as limiting the scope of the preceding Summary of the Invention in any way. The Detailed Description will make reference to a number of drawings as follows:

[0033] Figure 1 shows chemical formulae for two enantiomers of /;-menth-2-cn- 1 ,4-diol and the NMR spectrum.

[0034] Figure 2 is a perspective view of para-menthane-l, 2,4-triol, showing the labelling scheme and 30% thermal ellipsoids.

[0035] Figure 3 is a packing diagram for p « ra - m c n On an e - 1 , 2 , 4 - ( ri o I , viewed along the a axis.

[0036] Figure 4 is a table of the fractional atomic coordinates (xlO ⁴ ) and equivalent isotropic displacement parameters (A ² xl0 ³ ) for para-menthane- 1 ,2,4-triol.

[0037] Figure 5 is a table listing the anisotropic displacement parameters (A ² xl0 ³ ) for para- menthane- 1 ,2,4-triol.

[0038] Figure 6 is a table setting out bond lengths for para-menthanc- 1.2,4-triol .

[0039] Figure 7 is a table setting out bond angles for para-menthane- 1 , 2,4-triol.

[0040] Figure 8 is a table listing the hydrogen bonds for para-menthane-l, 2,4-triol.

[0041] Figure 9 is a table setting out the torsion angles for para-menthane- 1 , 2,4-triol.

[0042] Figure 10 is a table listing the hydrogen atom coordinates (AxlO ⁴ ) and isotropic displacement parameters (A ² xl0 ³ ) for para-menthane- 1 ,2,4-triol .

[0043] Figure 11 is a ¹³ C NMR spectrum of para- menthane-1, 2,4-triol.

[0044] Figure 12 is solid state NMR spectrum of para-menthane-l, 2,4-triol.

[0045] Figure 13 is a mass spectrum of para-menthane- 1 , 2,4-triol.

[0046] Figure 14 is a graph showing the anti-tumour activity of a composition of the invention on various cancers.

[0047] Figure 15A is a series of dose-response curves showing the effect of a composition of the invention on various cancers.

[0048] Figure 15B is a series of dose -response curves showing the effect of an alternative composition of the invention on various cancers.

[0049] Figure 16A is a table of in vitro test results of the effect of a composition of the invention on various cancer cell lines.

[0050] Figure 16B is a table of in vitro test results of the effect of an alternative composition of the invention on various cancer cell lines.

[0051] Figure 17 A is a series of graphs showing the anti-tumour activity of a composition of the invention on various cancer cell lines.

[0052] Figure 17B is a series of graphs showing the anti-tumour activity of an alternative composition of the invention on various cancer cell lines.

[0053] Figure 18A is a graphical overlay of the dose-response curves of Figure 15A, showing the effect of a composition of the invention on various cancers.

[0054] Figure 18B is a graphical overlay of the dose-response curves of Figure 15B, showing the effect of an alternative composition of the invention on various cancers.

[0055] Figures 19A and 19B are graphs of in vivo anti-tumour activity for a composition of the invention in a mouse xenograft model.

[0056] Figures 20A, 20B and 20C are graphs of in vivo anti- tumour activity for a composition of the invention in a mouse xenograft model.

DESCRIPTION OF EMBODIMENTS

[0057] A preferred composition of the invention comprises 2-15% p-menth-2-en- l ,4-diol. 0.5-25% (w/w) para-menthane - 1.2 , 4 - tr 8-30% (w/w) p-cymcnc. and 45-80% (w/w) terpinen- 4-ol.

[0058] More preferably, a composition of the invention comprises 3-10% p-menlh-2-en- 1 ,4- diol, 2-12% (w/w) /v/ra-menthane- l .2,4-triol, 10-20% (w/w) p-cymene, and 50-70% (w/w) terpinen-4-ol.

[0059] Even more preferably, a composition of the invention comprises 4-8% p-menth-2-en- 1,4-diol, 3-8% (w/w) para-menthane- 1 ,2,4-triol. 12-18% (w/w) p-cymene, and 52-68% (w/w) terpinen-4-ol.

[0060] Preparation of composition -“Sample 1”

[0061] Commercially available g-terpinene and terpinen-4-ol were combined in a 42:58 (v/v) ratio. The mixture was subjected to oxidation under conditions of high pressure (greater than atmospheric pressure) and elevated temperature (25 °C - 75 °C). The progress of the reaction was monitored by gas chromatography. When about 95% of the g-terpinene had converted to p-cymene, such that the ratio of p-cymene to terpinen-4-ol was about 1:4, the reaction was stopped by reducing the pressure back to atmospheric pressure.

[0062] The resultant mixture was stirred at atmospheric pressure and elevated temperature (35 °C - 55 °C) in the presence of oxidation for 6-12 hours. Analysis of the product (Sample 1) confirmed the presence of p-menth-2-en-l,4-diol and para-menthan- 1 ,2,4-triol.

[0063] Preparation of composition -“Sample 2”

[0064] Commercially available g-terpinene and terpinen-4-ol were combined in a 42:58 (v/v) ratio. The mixture was subjected to oxidation under conditions of high pressure (greater than atmospheric pressure) and elevated temperature (25 °C - 75 °C). The progress of the reaction was monitored by gas chromatography. When about 95% of the g-terpinene had converted to p-cymene, such that the ratio of p-cymene to terpinen-4-ol was about 1:4, the reaction was stopped by reducing the pressure back to atmospheric pressure.

[0065] The resultant mixture was stirred at atmospheric pressure and elevated temperature (35 °C - 55 °C) in the presence of oxidation for 12- 16 hours. Analysis of the product (Sample 2) confirmed the presence of p-menth-2-en-l,4-diol and para-mcmhan- 1 ,2,4-triol.

[0066] The final state of the composition can be in any suitable form, either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid form preparations include solutions, suspensions and emulsions. Either solid or liquid is suitable for oral ingestion.

[0067] For obtaining a solid product, microcrystalline cellulose was used as an excipient. Specifically, microcrystalline cellulose was mixed with a composition comprising p-menth-2-en- l,4-diol, para-menthane- 1 ,2,4-triol, p-cymcnc, and terpinen-4-ol, in a ratio of 2.5 : 1 (microcrystalline cellulose : composition) at 40 °C in a rotary oven over two hours.

[0068] Characterisation of p-menth-2-en- l ,4-diol

[0069] The presence of p-menth-2-en-l, 4-diol in compositions of the invention was verified by taking a composition of the invention, such as Sample 1, and subjecting to analysis by nuclear magnetic resonance (NMR). The NMR spectrum of two enantiomers of p-menth-2-en- l, 4-diol is shown in Figure 1.

[0070] Characterisation of para-menthane- 1 ,2.4-triol

[0071 ] The presence of para-menthane- 1 ,2,4-triol in compositions of the invention was verified by taking a composition of the invention, such as Sample 1, and specifically crystallizing from that composition, para-menthane-1, 2,4-triol. The specific crystallization of para-menthane- 1.2.4-triol from the composition was confirmed by X-ray diffraction (XRD), NMR and mass spectrometry (MS).

[0072] X-ray diffraction

[0073] A perspective view of para- men thane- 1.2,4-triol showing the labelling scheme and

30% thermal ellipsoids is shown in Figure 2.

[0074] A packing diagram for para-menthane-1, 2,4-triol viewed along the a axis is shown in Figure 3. Dashed lines represent the O FT O hydrogen bonding interactions.

[0075] The crystal data and structure refinement information for para-menthane- 1 ,2,4-triol is set out in Table II.

[0076] Table II - Crystal data for para-menthane-1, 2,4-triol

[0077] A table of the fractional atomic coordinates (xlO ⁴ ) and equivalent isotropic displacement parameters (A ² xl0 ³ ) for para-menthanc- 1 ,2.4-triol is set out in Figure 4. U _eq is defined as 1/3 of the trace of the orthogonalised Uu tensor.

[0078] A table listing the anisotropic displacement parameters (A ² xl0 ³ ) for para- menthane- 1,2,4-triol is set out in Figure 5. The anisotropic displacement factor exponent takes the form:

[0079] A table setting out bond lengths for para- me n t h a n e - 1 , 2 , 4 r i o 1 is shown in Figure 6.

[0080] A table setting out bond angles for para-menthane-l,2,4-triol is shown in Figure 7.

[0081] Figure 8 is a table listing the hydrogen bonds for para-menthane- 1 2,4-triol. [0082] Figure 9 is a table setting out the torsion angles for para- menthane-1, 2,4-triol.

[0083] A table listing the hydrogen atom coordinates (AxlO ⁴ ) and isotropic displacement parameters (A ² x10 ³ ) for para-menthane- 1 , 2,4-triol is shown in Figure 10.

[0084] A ¹³ C NMR spectrum of pctra-menthane- 1.2,4-triol is presented in Figure 11.

[0085] A solid state NMR spectrum of para- menthane-1, 2,4-triol is shown in Figure 12.

[0086] A mass spectrum of para-menthane-l, 2,4-triol is shown in Figure 13. The formula was calculated from the MS data as set out in Table III.

[0087] Table III - MS calculation

[0088] Characterisation of p-cvmene and terpinen-4-ol

[0089] p-cymene and terpinen-4-ol were purchased from Sigma-Aldrich to use as comparative standards. Gas chromatography (GC) of the purchased compounds compared to GC traces of Sample 1 after crystallization of para-menthane-1, 2,4-triol from Sample 1, confirmed that p-cymene and terpinen-4-ol remained in solution after crystallization of para-menthane- 1, 2,4-triol from Sample 1.

[0090] Example 1 - LD50 toxicity test

[0091] The aim of this study was to evaluate the oral toxicity of a composition of the invention on ICR mice and Albino Wistar rats.

[0092] Food grade microcrystalline cellulose was mixed with Sample 1 at a ratio of 2.5: 1 at 40 °C in a rotary oven for two hours.

[0093] A group of 10 male and 10 female ICR mice and a group of 10 male and 10 female Albino Wistar rats were selected at an age of 6 weeks. Animals were ranked by weight and sex and then randomly allocated into four groups, being two group of females (m=l0, n ₂ =10) and two group of males (m=10, n ₂ =10). The treatment groups met the regulatory requirements of having the No Observable Effects Limit (“NOEL”) group, the Lowest Observable Effects Limit (“LOEL”) group, and the Mean Dose Lethality (“MDL”) or Maximum Toxicity Dose (“MTD”) group.

[0094] During pre-test of MTD intragastrical administration, the animals appeared to show no sign of any effect. Intragastrical administration was 2850 mg/kg body weight over a period of 14 days for each group, with mortality being nil. There were no signs of intoxication, no difficulty in respiration, no signs of sedation or drowsiness, no change in locomotors activity, or lacrimation seen any group. All animals appeared healthy through the entire 14 day period. None of the animals in the test dose groups appeared to have been affected and therefore it was determined that there was no toxic effect when Sample 1 was administered at 2850 mg/kg body weight. The absence of toxicity is suggestive that Sample 1 has no adverse effect at LD ₅₀ 2850 mg.

[0095] Screening of Samples 1 and 2

[0096] Samples 1 and 2 were submitted to the National Cancer Institute Developmental Therapeutics Program for testing with the NCI-60, a panel of human tumour cell lines. The testing was performed according to NCI protocols, as described in Monks et al. (1991), J Nat Cancer Inst 83(11):757-766; Boyd and Pauli (1995), Drug Dev Res 34(2):91 - 109; Boyd (1997), Cancer Drug Discovery and Development. Teicher BA (Ed) Volume 2. Totowa, NJ, Human Press pp. 23-43, USA; and Shoemaker (2006) Nat Rev Cancer 6(10):813-823. The results of a single dose screening of Sample 1 are set out in Figure 14.

[0097] The single dose screening of Sample 1 indicated appreciable levels of anticancer activity, such that Samples 1 and 2 were subjected to further screening. Specifically, Samples 1 and 2 were tested in vitro against human tumour cell lines derived from nine neoplastic diseases, including leukemia, non-small cell lung cancer, colon cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer (see Figures 15A and 16A for Sample 1, and Figures 15B and 16B for Sample 2). From Figure 16B, which is data for Sample 2, it can be seen that a lethality of -100% was achieved for six cell lines in the panel, and a lethality of great than -90% was achieved for more than 20 cell lines in the panel.

[0098] Three dose-response parameters were obtained for Samples 1 and 2, and the data is set out in Figures 17A and 17B, respectively. Specifically, the dose-response parameters are (i) growth inhibition of 50% (GI ₅₀ ), (ii) total growth inhibition (TGI); and (iii) LC ₅₀ -

[0099] The GI ₅₀ is a growth-inhibitory effect, the TGI is a cytostatic level of effect, and the LC50 is the lethal concentration or cytotoxicity parameter. Where the tested parameters, logioGl50, logioTGI and logioLC ₅₀ , have negative log values of greater than -4.3, the Sample is deemed active. From the results set out in Figures 17A and 17B, it can be seen that both Samples showed significant growth inhibition, with the mean logioGLo for Sample 1 being -6.41, and for Sample 2 being -6.6.

[0100] Cell lines

[0101] Human LLC cells line were grown in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% heat-inactivated (HI) fetal bovine serum (FBS), 1% glutamine and streptomycin/penicillin.

[0102] MTT cell viability assay

[0103] Cells were seeded in 96-well plates (1x10 ⁴ cells/well) in complete medium. On the following day, different concentrations of the above-described agents were added to the cells in triplicates. At 72 hours, the medium was replaced by fresh media (100 mL per well) containing 1 mg/mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and incubated for 2-4 hours. MTT-formazan crystals were dissolved by the addition of 100 mL extraction buffer. Absorbance at 570 nm and a reference wavelength of 690 nm were recorded on an automated microplate reader. The relative number of viable cells was compared to untreated cells.

[0104] Transfection and luciferase assay

[0105] Transfections were performed using jet PEI™ (Polyplus-transfection Inc, NY, USA) according to the manufacturer’s instructions. Cells were seeded in 6-well plates for Luc assays. The next day, when the cells were about 50% confluent, co-transfection with3 pg of PY4-SV40- LUC vector plus 0.3 ng of pRL-CMV(Promega) was performed. 24 hours after the transfection medium was replaced and the cells were exposed to different concentrations of a composition of the invention, or left untreated. Luc assay was performed 48 hours post- treatment. Briefly, cells were washed once with phosphate buffered saline (PBS) and then lyzed in 250 mL of lysis buffer for 5 minutes at room temperature. The lysates were centrifuged at 10,000 rpm for 5 minutes, and 20 mL of each lysate were used to measure Luc reporter gene expression. The Luc activity was normalized to Renilla Luc activity from a parallel co-transfection of a pRL-CMVDual Luc system (Pro mega).

[0106] Detection of cell death

[0107] Cells were seeded in 12-well plates (1x10 ^s cells/well) in complete medium and treated with Sample 1 at high ( > 60%) and low (30-60%) concentrations for 72 hours. Annexin V was detected according to the manufacturer’s protocol. The cells were washed with PBS and then incubated in a solution of the membrane-impermeable nuclear dye propidium iodide. The cells were then immediately analyzed by flow cytometry [FACSCalibur (Becton Dickinson, CA)], and the results were analyzed with the CELL Quest program (Becton Dickinson). Cell death was achieved at both high (> 60%) and low (30-60%) concentrations of Sample 1.

[0108] Xenograft model for measuring in vivo anti-tumor activity of Sample 1

[0109] Intratumoral Injection

[0110] Male athymic nude mice, 6-8 weeks old, (Laboratories standard) were housed in sterile cages and handled with aseptic precautions. For testing the therapeutic potential of Sample 1, cells were implanted subcutaneously into the flank of the mice. When tumors were palpable (0.3-0.4 cm ³ ), the mice were randomly divided into four groups, each group having 8 members and the treatment was started (intratumoral injection).

[0111] The groups were treated as follows:

Group 1 - PBS

Group 2 - cyclophosamide (CP) intratumoral injection 40 mg/kg per tumor

Group 3 - compound having low activity with respect to depressing triglycerides and regulating the immune system (labelled as M5816 in Figures 19A and 19B), diluted with absolute alcohol to 10%, and 100 mL per tumor injected intratumorally

Group 4 - Sample 1 (labelled as M8182 in Figures 19 A and 19B), diluted with absolute alcohol to 10%, and 100 mL per tumor injected intratumorally

[0112] Animals were treated daily for 18 days. Tumor volume was measured with a caliper and plotted every day starting from the onset the treatment. Animals were anesthetized and sacrificed by cervical dislocation after 18 days.

[0113] The results are set out in Figures 19A and 19B. From the data, it can be seen that Sample 1 completely inhibited one tumor from Day 4, and completely inhibited the remaining tumours from Day 5. There was no indication of any of the tumors recurring.

[0114] Intraperitoneal Injection [0115] Male athymic C57LB/6 mice, 6-8 weeks old, were housed in sterile cages and handled with aseptic precautions. For testing the therapeutic potential of Sample 1, cells were implanted subcutaneously into the flank of the C57LB/6 mice. When tumors were palpable (0.3-0.4 cm ³ ), the mice were randomly divided into four groups, each group having 8 members and the treatment was started (intraperitoneal injections).

[0116] The groups were treated as follows:

Group 1 - PBS

Group 2 - cyclophosamide (CP) intraperitoneal injection 40 mg/kg per mouse

Group 3 - Sample 1 was diluted with DMSO to give a solution of 0.4% (labelled as M5816 in Figures 20A, 20B and 20C), and 1.25 mL/kg per mouse was injected intraperitoneally

Group 4 - Sample 1 was diluted with DMSO to give a solution of 0.4% (labelled as M8182 in Figures 20A, 20B and 20C), and 2.5 mL/kg per mouse was injected intraperitoneally

[0117] The results are set out in Figures 20A, 20B and 20C. From the data, it can be seen that Sample 1 completely inhibited all but two tumors from Day 6, and completely inhibited all tumours from Day 7. There was no indication of any of the tumors recurring.

[0118] In summary, a composition of the invention, comprising a combination of p-menth- 2-en-l,4-diol, para-menthane-l,2,4-triol, p-cymene, and terpinen-4-ol shows activity in treating lung cancer, in particular Lewis lung carcinoma.

[0119] In the present specification and claims (if any), the word‘comprising’ and its derivatives including‘comprises’ and‘comprise’ include each of the stated integers but does not exclude the inclusion of one or more further integers.

[0120] Reference throughout this specification to‘one embodiment’ or‘an embodiment’ means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearance of the phrases‘in one embodiment’ or‘in an embodiment’ in various places throughout this specification are not necessarily all referring to the same embodiment.

Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more combinations. [0121] In compliance with the statute, the invention has been described in language more or less specific to structural or methodical features. It is to be understood that the invention is not limited to specific features shown or described since the means herein described comprises preferred forms of putting the invention into effect. The invention is, therefore, claimed in any of its forms or modifications within the proper scope of the appended claims (if any) appropriately interpreted by those skilled in the art.