WO2023238114A1

WO2023238114A1 - Combinations and methods for the treatment of cancer

Info

Publication number: WO2023238114A1
Application number: PCT/IL2022/050612
Authority: WO
Inventors: Hinanit KOLTAI; Nurit SHALEV
Original assignee: The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Institute)
Priority date: 2022-06-08
Filing date: 2022-06-08
Publication date: 2023-12-14

Abstract

The present disclosure provides combinations of CBC and THC for use in reducing cell viability and treating cancer.

Description

COMBINATIONS AND METHODS FOR THE TREATMENT OF CANCER

TECHNOLOGICAL FIELD

The present disclosure relates to combinations, compositions and methods for the treatment of cancer.

BACKGROUND ART

References considered to be relevant as background to the presently disclosed subject matter are listed below:

- Ramer, R.; Hinz, B. Cannabinoids as anticancer drugs. In, Advances in Pharmacology 80, ed. by Kendall, D; S.P.H. Alexander (Elsevier Academic Press, 2017), 2017, pp. 397-436

- Velasco, G.; Hernandez-Tiedra, S.; Davila, D.; Lorente, M. The use of cannabinoids as anticancer agents. Prog. Neuropsychopharmacol. Biol. Psychiatry 2016, 64, 259-266

- Anis, O., Vinayaka, A.C., Shalev, N., Namdar, D., Nadarajan, S., Anil, S.M., Cohen, O., Belausov, E., Ramon, J., Mayzlish Gati, E. and Koltai, H., Cannabis-Derived Compounds Cannabichromene and A9-Tetrahydrocannabinol Interact and Exhibit Cytotoxic Activity against Urothelial Cell Carcinoma Correlated with Inhibition of Cell Migration and Cytoskeleton Organization. Molecules, 2021, 26(2), p.465.

- Shalev N., Kendall K., Anil SM., Vinayaka AC., and Koltai, H., A9- tetrahydrocannabinol (THC)-rich compositions from cannabis have cytotoxic activity against ovarian cancer cells and act synergistically with niraparib in vitro, 2021

Acknowledgement of the above references herein is not to be inferred as meaning that these are in any way relevant to the patentability of the presently disclosed subject matter. BACKGROUND

Studies using Cannabis sativa have shown that phytocannabinoids possess anticancer activities including in skin, prostate, pancreatic, bladder, lung, breast, and glioma cancer cells.

These activities include cytotoxicity, induction of apoptosis, inhibition of cell proliferation and angiogenesis and inhibition of cell migration (Ramer, R. et al).

THC was previously suggested to have anticancer activity for a variety of cancer types, whereas CBC was mostly associated with anticancer activity on prostate carcinomas (Velasco, G. et al).

Moreover, THC and CBC were shown to interact to confer cytotoxicity against urothelial carcinoma cells (Anis, O. et al).

Further, it was shown that THC, CBC with other cannabinoids, confer high level of cytotoxic activity against ovarian cancer cells (Shalev N. et al.)

GENERAL DESCRIPTION

The present disclosure provides in accordance with some aspects, a combination comprising CBC and THC for use in reducing cell viability, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The present disclosure provides in accordance with further aspects, a combination comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The present disclosure provides in accordance with some aspects, a pharmaceutical composition comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL. The present disclosure provides in accordance with some aspects, a method of reducing cell viability, the method comprising contacting the cell with an effective amount of a composition comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The present disclosure provides in accordance with some aspects, a method of treating, preventing, or delaying the onset of cancer in a subject in need thereof, the method comprising administering to the subject a combination comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The present disclosure provides in accordance with some aspects, a kit comprising CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and optionally comprising instructions for using said CBC and said THC in a combination for beating, preventing, or delaying the onset of cancer in a subject in need thereof.

The present disclosure provides in accordance with some aspects, use of CBC and THC in the preparation of a pharmaceutical composition for heating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said pharmaceutical composition comprises CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL

BRIEF DESCRIPTION OF THE DRAWINGS

In order to better understand the subject matter that is disclosed herein and to exemplify how it may be carried out in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which:

Figs. 1A-1H are graphs of cell viability following treatment with THC or CBC at different concentrations for IC50 value calculation, Figs. 1A and Fig. IB - IC50 calculation of THC and CBC, respectively in HTB161 cells; Figs. 1C and Fig. ID - IC50 calculation of THC and CBC, respectively in HTB75 cells; Figs. IE and Fig. IF - IC50 calculation of THC and CBC, respectively in A549 cells; Figs. 1G and Fig. 1H - IC50 calculation of THC and CBC, respectively in HCT116 cells; results obtained from 5P logistic curve fit using GraphPad Prism version 6.1, error bars indicate ± SE (n=3).

Figs. 2A-2C show synergistic interactions between THC and CBC on cell viability on HTB75 cells, Fig. 2A at 24 hours, Fig. 2B at 48 hours, Fig. 2C at 72 hoursfollowing combined treatments, synergy of cytotoxic activity calculated based on the Bliss independence drug interaction model; delta value of observed values minus expected values are shown in the Y axis, HSD; P < 0.05, for the synergistic delta values of THC and CBC on the cell line.

Figs. 3A-3C show synergistic interactions between THC and CBC on cell viability in HTB161 cells, Fig. 3A at 24 hours, Fig. 3B at 48 hours, Fig. 3C at 72, following combined treatments, synergy of cytotoxic activity calculated based on the Bliss independence drug interaction model; delta value of observed values minus expected values are shown in the Y axis, HSD; P < 0.05, for the synergistic delta values of THC and CBC on the cell line.

Fig. 4 show synergistic interactions between THC and CBC on cell viability at 48h of A549 cells following combined treatments, synergy of cytotoxic activity calculated based on the Bliss independence drug interaction model; delta value of observed values minus expected values are shown in the Y axis, n=3; HSD; P < 0.05, for the synergistic delta values of THC and CBC on the cell line.

Fig. 5 show synergistic interactions between THC and CBC on cell viability at 48h of HCT116 cells following combined treatments, synergy of cytotoxic activity calculated based on the Bliss independence drug interaction model; delta value of observed values minus expected values are shown in the Y axis, n=3; HSD; P < 0.05, for the synergistic delta values of THC and CBC on the cell line.

DETAILED DESCRIPTION OF EMBODIMENTS

The present disclosure is based on the findings that combinations of cannabinoids, including, inter alia, cannabichromene (CBC) and tetrahydrocannabinol (THC) are effective in reducing cell viability of various cell types, specifically various cancers. As shown in the Examples below, various combinations of CBC and THC had an effect and specifically a synergistic effect in reducing cell viability in a variety of cancers. This reduced cell viability was superior compared to the effect of each one of the cannabinoids (THC and CBC) on cell viability when administered as single compounds.

As also shown in the Examples below, this effect was observed in a broad range of CBC and THC concentrations and concentrations ratios in multiple cancer types.

Hence, it was suggested that CBC and THC can interact synergistically to confer cell death in different cancer cells.

Surprisingly, the Examples below show that synergism was observed in a broad range of CBC to THC combinations, including, inter alia, combinations in which CBC is in excess as compared to THC and combinations in which THC is in excess as compared to CBC.

Thus, the present disclosure provides a synergistic combination comprising CBC and THC for use in reducing cell viability.

In accordance with some aspects, the present disclosure provides, a combination comprising CBC and THC for use in reducing cell viability, wherein CBC is present at a concentration of at least about 0.5 pg/mL and THC is present at a concentration of at least about 0.5 pg/mL.

The present disclosure also provides, by some aspects a combination comprising CBC and THC for use in reducing cell viability, wherein CBC is present at a concentration of at least about 0.5 pg/mL and about 30 pg/mL, wherein THC is present at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

Cell viability is a measure of live cells and is often defined as the percentage of living cells in a sample.

Cell viability can be correlated with cell proliferation, as both measure live cells with proliferation also being a measure of the rate of growth and production of daughter cells of a cell population.

In some embodiments, the combination is for use in inducing cell cycle arrest and/or reducing migration of cancer cells and/or reducing/inhibiting cell proliferation. In some other embodiments, the cell is obtained from a subject suffering from cancer or who is predicted to suffer from cancer or who has been diagnosed to potentially develop cancer.

Based on the above detailed findings, it was suggested by the inventors that combinations of these cannabinoids can be used in cancer therapy.

Thus, the present disclosure provides a synergistic combination comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof.

In accordance with another aspect of the invention, the present disclosure provides a combination comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination comprises CBC at a concentration of at least about 0.5 pg/mL and THC at a concentration of at least about 0.5 pg/mL.

In some embodiments, the combination for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof comprises CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In the following text, when referring to a combination it is to be understood as also referring to a composition, pharmaceutical composition, kit, uses or methods disclosed herein. Thus, whenever providing a feature with reference to the combination, it is to be understood as defining the same feature with respect to the compositions, pharmaceutical compositions, kits, uses and methods, mutatis mutandis.

Thus, the present invention also provides combinations, compositions, , kits, uses and methods as herein defined and exemplified.

As described the combination comprises cannabinoids, including, inter alia, CBC and THC.

As used herein, the term cannabinoid refers to compounds found in cannabis plant as well as to synthetic compounds that are capable of interacting with an endocannabinoid system. The term cannabichromene (CBC) (also denoted as cannabichrome, cannanbichromene, pentylcannabichromene or cannabinochromen), having IUPAC name 2-methyl-2-(4-methylpent-3-enyl)-7-pentyl-5-chromenol and CAS Number 20675-51-8 as used herein encompasses a naturally occurring compound, synthetic compound, semisynthetic compound or analogues thereof.

Non-limiting examples of CBC analogs include isocannabichromene, cannabichromene-cO, cannabichromene-cl, isocannabichromene-cO, CBCan.

Tetrahydrocannabinol (THC) having IUPAC name (6a/?,10aR)-6,6,9-Trimethyl-

3-pentyl-6a,7,8,10a-tetrahydro-6Z7-benzo[c]chromen-l-ol and CAS Number 1972-08-3 as used herein encompasses a naturally occurring compound, synthetic compound, semisynthetic compound or analogues thereof.

Non-limiting examples of THC analogs include (-)-A9-trans- Tetrahydrocannabinol-C5 (A9-THC), (-)-A8-trans-(6aR,10aR)-Tetrahydrocannabinol-C5 (A8-THC), A8-THC-C2, A9-trans-Tetrahydrocannabivarin A9-THCV (THCV), (-)-A8-trans- THC-C3 (A8-THCV), A7-tetrahydrocannabivarin-C3 (A7-THCV), (-)-A9-trans- Tetrahydrocannabinol-C4 (A9-THC-C4), (-)-trans-A8-THC-C4, (-)-A9-trans- Tetrahydrocannabinol-C5 (A9-THC), (-)-A8-trans-(6aR,10aR)- Tetrahydrocannabinol-C5 (A8- THC), (-)-A9-(6aS,10aR-cis)-Tetrahydrocannabinol-C5 ((-)-cis-A9-THC), (-)-A7-trans- (1R,3R,6R)-Isotetrahydrocannabinol-C5 (trans-iso-A7-THC).

At times, THC refers to the delta-9-THC isomer having a chemical name (■■)■ /raris- A⁹-tetrahydrocannabinol.

Reference to a CBC analog should be understood as a compound having the effect of CBC as described herein, either alone or in combination with THC. In other words, a CBC analogue has an anti-cancer activity as described herein and specifically a synergistic effect as described herein.

Reference to a THC analog should be understood as a compound having the effect of THC as described herein, either alone or in combination with CBC. In other words, a THC analogue has an anti-cancer activity as described herein and specifically a synergistic effect as described herein.

In accordance with the present disclosure, CBC and THC can be obtained from any possible source.

In some embodiments, CBC is a naturally obtained CBC. In some embodiments, THC is a naturally obtained THC.

Naturally occurring CBC and/or THC can be obtained from an extract of a cannabis plant.

In some embodiments, at least one of the CBC and/or the THC is obtained from an extract of a cannabis plant.

The cannabis plant is not limited and, in some embodiments, the extract is obtained from any one of sativa, indica, or ruderalis species. It should be noted that when both CBC and/or THC are obtained naturally obtained, each one can be obtained from a different cannabis plant, or both can be obtained from the same cannabis plant.

The “extract” (also referred to as "concentrate"') obtained from the cannabis plant is a product of separation (or isolation) of one or more substances from a plant or a plant material using a solvent, or by using any other extraction technique known in the art. The extract may be obtained by extraction the plant material with water or a water containing solvent, or from a non-aqueous system comprising at least one suitable solvent, e.g., pentane, decane, cyclohexane, hexane, petroleum ether, chloromethane, ethanol, butanol, acetone, dichloromethane, chloroform, isopropanol, propanol, ethyl acetate, methanol, butylene glycol, propylene glycol, pentylene glycol, glycerol, ethers, oils, water or any other suitable solvent, and any mixture of these solvents. Extraction may be done by any conventional method in the art or such methods specifically suitable for extracting cannabis components.

It should be noted that in accordance with some embodiments in which at least one of the CBC and/or the THC is obtained from a cannabis plant, the combination may comprise other cannabinoids that are obtained together with the at least one of the CBC and/or the THC.

In other words, as the cannabis plant extract may be obtained by, e.g., extraction directly from a cannabis plant, it can include in addition to CBC and THC, at least one natural cannabinoid, i.e., cannabidiol (CBD), cannabigerol (CBG), cannabicyclol (CBL), CBV (cannabivarin), THCV (tetrahydrocannabivarin), CBDV (cannabidivarin), CBCV (cannabichrome varin), CBGV (cannabigerovarin) and CBGM (cannabigerol monomethyl ether).

It should be appreciated that the material composition and other properties of the extract may vary and further may be tailored to meet the desired properties of a combination according to the invention.

In some embodiments, CBC is not naturally occurring CBC.

In some embodiments, THC is not naturally occurring THC.

In some embodiments, at least one of the CBC and/or the THC is synthetically produced. As appreciated, the at least one of the CBC and/or the THC can be synthesized by any known method in the art.

In some embodiments, the combination is a synthetic combination.

In some embodiments, CBC is not synthetically produced CBC.

In some embodiments, THC is not synthetically produced THC.

In some embodiments, the combination does not comprise cannabidiol (CBD).

In some embodiments, the combination does not comprise cannabigerol (CBG). In some embodiments, the combination does not comprise cannabicyclol (CBL).

In some embodiments, the combination does not comprise CBV (cannabivarin).

In some embodiments, the combination does not comprise THCV (tetrahydrocannabivarin). In some embodiments, the combination does not comprise CBDV (cannabidivarin). In some embodiments, the combination does not comprise CBCV (cannabichromevarin). In some embodiments, the combination does not comprise CBGV (cannabigerovarin). In some embodiments, the combination does not comprise CBGM (cannabigerol monomethyl ether). In some embodiments, the combination is free of cannabidiol (CBD), and cannabigerol (CBG).

In some embodiments, the combination comprises CBC and THC as major components. In some embodiments, the combination comprises at least about 50% CBC and THC, at least about 60% CBC and THC, at least about 70% CBC and THC, at least about 80% CBC and THC, at least about 90% CBC and THC, at times at least about 95% CBC and THC.

In accordance with the present disclosure, a combination being free of a substance also referred to as not comprising a substance, also encompasses a combination comprising said substance in an amount that can not be detected by methods known in the art, i.e. below a detection level.

In some embodiments, the combination comprises two cannabinoids, the two cannabinoids being CBC and THC. In other words, while the combination can comprise additional compounds, in addition to CBC and THC, none of them is a cannabinoid.

In some embodiments, the combination consists of CBC and THC.

As shown herein, the examples demonstrate multiple active combinations of CBC and THC at varying concentrations and varying ratios of the two cannabinoids. The combination described herein may be considered as a therapeutic combination.

In some embodiments, the combination comprises the CBC at a concentration of between about 0.5 pg/mL and about 25 pg/mL and the THC at a concentration of between about 0.5 pg/mL and about 30pg/mL, at times THC at a concentration of between about 0.5 pg/mL and about 25 pg/mL.

It should be noted that when referring to a combination of a range of CBC concentrations and a range of THC concentrations, it encompasses all possible combinations comprising each CBC concentration and each THC concentration. For example, a combination comprises CBC at a concentration of between about 1 pg/mL and about 1.5 pg/mL and THC at a concentration of between about 1 pg/mL and about 1.5 pg/mL, encompasses a matrix of concentrations such that the combination comprises CBC at any concentration between about 1 pg/mL and about 1.5 pg/mL with THC at any concentration between about 1 pg/mL and about 1.5 pg/mL. In other words, any value which is between any one of the values and/or any range between one or more of the above values is within the scope of the present disclosure.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 20 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In some embodiments, the combination comprises the CBC at a concentration of between about 0.5 pg/mL and about 16 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 17.5 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 17.5 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 17.5 pg/mL. In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 14 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 14 pg/mL, at times THC at a concentration of between about 0.7 pg/mL and about 14 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 14 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 14 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 12.5 pg/mL, at times THC at a concentration of between about 0.7 pg/mL and about 12.5 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 12.5 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 12.5 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 10 pg/mL and THC at a concentration of between about 10 pg/mL and about 20 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 1 pg/mL and about 25 pg/mL and THC at a concentration of between about 1 pg/mL and about 25 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 1 pg/mL and about 20 pg/mL and THC at a concentration of between about 1.1 pg/mL and about 25 pg/mL, at times THC at a concentration of between about 2.5 pg/mL and about 25 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 25 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 1 pg/mL and about 20 pg/mL and THC at a concentration of between about 5 pg/mL and about 15 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 5 pg/mL and about 15 pg/mL and THC at a concentration of between about 10 pg/mL and about 15 pg/mL.

In some embodiments, the combination comprises CBC at a concentration of between about 5 pg/mL and about 25 pg/mL and THC at a concentration of between about 1 pg/mL and about 25 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 20 pg/mL.

In some embodiments, the combination comprises CBC and THC, each one of CBC and THC in at least one concentration according to Table 1.

It should be noted that each combination of CBC and THC in Table 1 (for example CBC at 5 pg/mL with THC at 15 pg/mL) is considered a combination in accordance with the present disclosure. Additionally, each combination of CBC and THC in Table 1 (for example CBC at 5 pg/mL with THC at 15 pg/mL) is considered as an embodiment of the invention. This applies mutatis mutandis to each one of Tables mentioned herein below and those which are provided in the Examples below, all form part of the disclosure.

In some embodiments, the combination comprises CBC and THC, each in at least one concentration according to any one of Table 2A, Table 2B, Table 3, Table 4, Table 5 or any combination thereof.

As cab be seen, Table 1, Table 2A, Table 2B, Table 3, Table 4 and Table 5 provides multiple combinations of CBC and THC such that each one of the rows in these Tables correspond to different concentrations of CBC and each one of the column in these Tables corresponds to different concentrations of THC. An active combination as used herein is or comprises a combination that is considered as having an effect in reducing cell viability and/or treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof. This active combination is marked in any one of Table 1, Table 2 A, Table 2B, Table 3, Table 4 and Table 5 with a

It should be noted that the present disclosure also encompasses combinations having CBC to THC ratio as described in any one of Table 1, Table 2A, Table 2B, Table 3, Table 4 and Table 5. For example, in an active combination comprises CBC at 12 pg/mL and THC at 6 pg/mL, the CBC to THC ratio is 2 and hence the present disclosure encompasses combinations in which according to this specific example the CBC to THC ratio is 2. The same applies to all combinations being active combinations in any one of Table 1, Table 2 A, Table 2B, Table 3, Table 4 and Table 5.

In some embodiments, the combination comprises CBC and THC in a concentration and a concentration ratio according to Table 2A, Table 2B or combination thereof. In some embodiments, the combination is at least one active combination as defined in Table 2 A, Table 2B or combination thereof.

In some embodiments, the combination comprises CBC and THC in a concentration and a concentration ratio according to Table 3. In some embodiments, the combination is at least one active combination as defined in Table 3.

In some embodiments, the combination comprises CBC and THC in a concentration and a concentration ratio according to Table 4. In some embodiments, the combination is at least one active combination as defined in Table 4.

In some embodiments, the combination comprises CBC and THC in a concentration and a concentration ratio according to Table 5. In some embodiments, the combination is at least one active combination as defined in Table 5.

As described herein, the combinations of CBC and THC have a synergistic effect and specifically a synergistic effect on cancer cell viability.

The term synergistic effect or synergistic activity are interchangeably herein interchangeably.

In some embodiments, the combination comprising CBC and THC has a synergistic activity as compared to each of said CBC and said THC when administered individually.

Hence, the combination comprising CBC and THC comprising CBC and THC may be considered as a synergistic combination.

A synergistic combination as used herein refers to a quantity of a combination comprising CBC and THC that is statistically significantly more effective/active than the additive effects of CBC and THC when used individually (e.g. not in combination).

A “synergistic amount” for a component in a combination (e.g. CBC and THC) is defined as an amount (e.g. concentration) providing a synergistic effect.

In some embodiments, each one of CBC and THC is present in the combination in a concentration capable of providing a synergistic effect. In other words, and in accordance with some other embodiments, each one of CBC and THC is present in the combination in a synergistic amount. As detailed herein and shown in the Examples, the synergistic amount may be provided by a range of concentrations for both CBC and THC, such that various combinations of CBC and THC provide a synergistic effect.

Hence, in accordance with the present disclosure, the synergistic amount of CBC may be a range of CBC concentrations or may be a single CBC concentration.

Further and in accordance with the present disclosure, the synergistic amount of THC may be a range of THC concentrations or may be a single THC concentration.

A determination of synergism between CBC and THC may be based on the results obtained from the methods described herein, specifically methods described in the Examples below.

For example, the synergistic effect of CBC and THC on cell viability can be determined using any known assay known in the art, including, cell viability assay described in the Examples below.

In some embodiments, the synergistic effect of CBC and THC on cell viability is determined as a synergistic combination using a Bliss independence drug interaction model method.

In some other embodiments, the synergistic effect of CBC and THC on cell viability is determined as a synergistic combination using a Loewe additivity model method.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC, wherein each CBC is present in the combination a synergistic amount and THC is present in the combination a synergistic amount.

As appreciated, each one of CBC and THC may have the same synergistic amount (either a range of concentrations or a single concentration as shown herein) or a different synergistic amount (either a range of concentrations or a single concentration as shown herein). Additionally, or alternatively, in case the synergistic amount is range of concentrations, the ranges may partially overlap.

For example, Table 1 may be considered as showing synergistic amount of CBC and synergistic amount THC in the combination, as provided by their concentrations (range of concentrations) and the corresponding ratio. It should be noted that refence to CBC and THC concentrations in Table 1 may be considered as±5%, at times according ±10%. The same applied mutatis mutandis to each one of Table 2A, Table 2B, Table 3, Table 4 and Table 5.

As shown in the Examples below, the combinations of the invention exhibited an effect and specifically a synergistic effect in reducing cell viability in various cancer cells.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability the combination, wherein CBC is present in the combination in a synergistic amount and THC is present in the combination in a synergistic amount.

The cancer cell can be a cell obtained from a subject suffering from cancer or a subject who is predicted to suffer from cancer or a subject who has been diagnosed to potentially develop cancer.

Hence, it was suggested that the combinations of the present disclosure can be beneficial in beating, preventing, or delaying the onset of cancer in a subject in need thereof.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein each CBC is present in the combination a synergistic amount and THC is present in the combination a synergistic amount.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 1.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 2 A.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 2B.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 3.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 4.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 5.

In some embodiments, the subject in need thereof is a subject suffering from cancer or a subject who is predicted to suffer from cancer or a subject who has been diagnosed to potentially develop cancer.

Cancer as defined herein is a disorder displaying cell division and growth that is not part of normal cellular turnover, metabolism, growth, or propagation of the whole organism. Unwanted proliferation of cells is seen in tumors and other pathological proliferation of cells, does not serve normal function, and for the most part will continue unbridled at a growth rate exceeding that of cells of a normal tissue in the absence of outside intervention. A pathological state that ensues because of the unwanted proliferation of cells is referred herein as a "hyper-proliferative disease" or "hyper- proliferative disorder." It should be noted that the term “proliferative disorder”, “cancer”, “tumor” and “malignancy” all relate equivalently to a hyperplasia of a tissue or organ.

It should be noted that the term cancer when used herein encompasses non- invasive cancer, invasive cancer and metastatic cancer.

"Non-invasive" cancer is to be understood as a cancer that do not grow into or invade normal tissues within or beyond the primary location, for example the ovary.

"Invasive cancers" is to be understood as cancer that invades and grows in normal, healthy tissues to form metastasis.

"Metastatic cancer" or "metastatic status" is to be understood as a cancer that has spread from the place where it first started to another place in the body. Such a tumor formed by metastatic cancer cells is called a metastatic tumor or a metastasis.

Malignancy encompasses any one of carcinomas, melanomas, lymphomas, leukemias, myeloma and sarcomas.

Non-limiting examples of cancers include blastoma, carcinoma, lymphoma, leukemia, sarcoma, mesothelioma, glioma, germinoma, choriocarcinoma, melanoma, glioblastoma, lymphoid malignancies, squamous cell cancer (e.g. epithelial squamous cell cancer), lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, esophageal cancer as well as head and neck cancer.

In some embodiments, the cell is not selected from the group consisting of urothelial cancer, uterine cervix carcinoma, prostate cancer and melanoma. In some embodiments, the cell is not urothelial cancer cell.

In some embodiments, the cell is not a glioblastoma cell.

In some embodiments, the cancer may be any one of a carcinoma.

The term "carcinoma" refers herein to any tumor tissue derived from putative epithelial cells, or epithelial tissue that has glandular origin, and in many cases are associated with epithelial-mesenchymal transition (EMT).

In some embodiments, the cancer is not selected from the group consisting of urothelial cancer, uterine cervix carcinoma, prostate cancer and melanoma.

In some embodiments, the cancer is not urothelial cancer.

In some embodiments, the cancer is not glioblastoma.

In some embodiments, the cancer is a solid tumor.

In some embodiments, the cancer is selected from the group consisting of ovarian cancer, lung cancer and colon cancer.

In some embodiments, the cancer is lung cancer or colon cancer.

In some embodiments, the cancer is ovarian cancer.

The term ovarian cancer as referred herein encompasses several different types of cancers that arise from cells of the ovary, including, inter alia, epithelial ovarian carcinomas, germ cell tumors, and stromal cell tumors.

In some embodiments, the combination has a synergistic activity against ovarian cancer and specifically reducing cell viability in ovarian cancer cells

Hence, in accordance with some aspects, the present disclosure provides a combination comprising a synergistic amount of CBC and a synergistic amount of THC, for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the concentration of CBC in the combination is at least about 0.5% (w/w), at times at least about 1%, at times at least about 2%, at times at least about 3%, at times at least about 4%, at times at least about 5%, at times at least about 6%, at times at least about 10%, at times at least about 15%, at times at least about 20%, at times at least about 25%, at times at least about 30%, at times at least about 35%, at times at least about 40%, at times at least about 45%, at times at least about 50%, at times at least about 55%, at times at least about 60%, at times at least about 65%, at times at least about 70%, at times at least about 80%, at times at least about 85%, at times at least about 90%, at times at least about 95%. Any value which is between any one of the above values and/or any range between one or more of the above values is within the scope of the present disclosure.

In some embodiments, the concentration of CBC in the combination is between about 0.5% and about 97%.

In some embodiments, the concentration of THC in the combination is at least about 0.5% (w/w), at times at least about 1%, at times at least about 2%, at times at least about 3%, at times at least about 4%, at times at least about 5%, at times at least about 6%, at times at least about 10%, at times at least about 15%, at times at least about 20%, at times at least about 25%, at times at least about 30%, at times at least about 35%, at times at least about 40%, at times at least about 45%, at times at least about 50%, at times at least about 55%, at times at least about 60%, at times at least about 65%, at times at least about 70%, at times at least about 80%, at times at least about 85%, at times at least about 90%, at times at least about 95%. Any value which is between any one of the above values and/or any range between one or more of the above values is within the scope of the present disclosure.

In some embodiments, the concentration of THC in the combination is between about 0.5% and about 97%.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 16 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 17.5 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 17.5 pg/mL, at times THC at a concentration of between about 5pg/mL and about 17.5 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 14 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 14 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 14 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 14 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 12.5 pg/mL, at times THC at a concentration of between about 1 pg/mL and about 12.5 pg/mL, at times THC at a concentration of between about 5 pg/mL and about 12.5 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and THC at a concentration of between about 5 pg/mL and about 15.5 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and THC at a concentration of between about 5pg/mL and about 12.5pg/mL, (ii) CBC at a concentration of about 8 pg/mL and THC at a concentration of about 0.5 pg/mL; or (iii) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of about 12.5pg/mL, (ii) CBC at a concentration of between about 4 pg/mL and about 12 g/mL and THC at a concentration of about 10 pg/mL and about 12.5 pg/mL; (iii) CBC at a concentration of between about 8 pg/mL and about 12 g/mL and THC at a concentration of about 5 pg/mL and about 12.5pg/mL or (iii) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and THC at a concentration of between about 5pg/mL and about 12.5pg/mL. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and 8 pg/mL THC at a concenhation of between about 7.5pg/mL and about 12.5pg/mL, (ii) CBC at a concentration of between about 4 pg/mL and about 8 g/mL and THC at a concentration of about 5 pg/mL and about 10 pg/mL; (iii) CBC at a concentration of about 8 pg/mL and THC at a concentration of about 0.5 pg/mL and about lOpg/mL or (iii) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of between about 7.5pg/mL and about lOpg/mL (ii) CBC at a concentration of about 4 pg/mL and THC at a concentration of between about 5 pg/mL and about 10 pg/mL (iii) CBC at a concentration of about 8 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 10 pg/mL; or (iv) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of about 0.5 pg/mL and THC at a concenhation of about 30 pg/mL, (ii) about 5 pg/mL and THC at a concenhation of between about 20 pg/mL and about 30 pg/mL (iii) CBC at a concentration of about 10 pg/mL and THC at a concentration of between about 10 pg/mL and 30 pg/mL, (iv) CBC at a concentration of between about 15 pg/mL and about 20 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL, or (v) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 5 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 25 pg/mL (ii) CBC at a concentration of about 0.5 pg/mL and THC at a concentration of between about 15 pg/mL and 30 pg/mL, (iii) CBC at a concentration of between about 15 pg/mL and about 20 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 5 pg/mL, (iv) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 0.5 pg/mL or (v) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of 0.5 pg/mL and THC at a concentration of between about 15 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 10 pg/mL and 15 pg/mL, (iii) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 10 pg/mL or (iv) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and 5 pg/mL and THC at a concentration of between about 15 pg/mL and about 20 pg/mL, (ii) CBC at a concentration of about 10 pg/mL and THC at a concentration of between about 10 pg/mL and about 20 pg/mL (iii) CBC at a concentration of about 15 pg/mL and THC at a concentration of between about 5 pg/mL and 15 pg/mL, or (iv) any combinations thereof. In such embodiments, t such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof. In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of 4 pg/mL a and THC at a concentration of between about 5 pg/mL and about 10 pg/mL, (ii) CBC at a concentration of 8 pg/mL a and THC at a concentration of between about 0.5 pg/mL and about 12.5 pg/mL, (iii) CBC at a concentration of 12 pg/mL a and THC at a concentration of between about 5 pg/mL and about 12.5 pg/mL, or (iv) any combinations thereof. In such embodiments, such a combination is a synergistic combination is for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of (i) CBC at a concentration of between about 0.5 pg/mL and THC at a concenhation of 5 pg/mL, (ii) CBC at a concentration of between about 0.5 pg/mL and THC at a concenhation of 7.5 pg/mL (iii) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of 10 pg/mL, (iv) CBC at a concenhation of between about 0.5 pg/mL and THC at a concenhation of 12.5 pg/mL, (v) CBC at a concenhation of between about 4 pg/mL and THC at a concentration of 5 pg/mL, (vi) CBC at a concentration of between about 4 pg/mL and THC at a concentration of 7.5 pg/m, (vii) CBC at a concentration of between about 4 pg/mL and THC at a concenhation of 10 pg/m, (viii) CBC at a concentration of between about 4 pg/mL and THC at a concenhation of 12.5 pg/m, (ix) CBC at a concenhation of between about 8 pg/mL and THC at a concenhation of 0.5 pg/m, (x) CBC at a concentration of between about 8 pg/mL and THC at a concentration of 5 pg/m, (xi) CBC at a concentration of between about 8 pg/mL and THC at a concentration of 7.5 pg/m, (xii) CBC at a concentration of between about 8 pg/mL and THC at a concentration of 10 pg/m, (xiii) CBC at a concentration of between about 8 pg/mL and THC at a concentration of 12.5 pg/m, (xiv) CBC at a concentration of between about 12 pg/mL and THC at a concentration of 5 pg/mL , (xv) CBC at a concentration of between about 12 pg/mL and THC at a concentration of 7.5 pg/m, (xvi) 12: 10, (xvii) 12: 12.5 or (xviii) any combination thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof. In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of about 12.5pg/mL (ii) CBC at a concentration of about 4 pg/mL and THC at a concentration of between about 10 pg/mL and about 12.5 pg/mL (iii) CBC at a concentration of between about 8 pg/mL and about 12.5 pg/mL and THC at a concentration of between about 5 pg/mL and about 12.5 pg/mL; or (iv) any combinations thereof. In such embodiments, such a combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or for use in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination comprises between about 3% and about 96% CBD for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

It should also be noted, that a synergism combination in accordance with the present disclosure encompasses one or more combinations of CBC concentrations and THC concentrations, from Table 1 as well as other Tables provided herein including those in the Examples below.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a synergistic combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 2 A.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a synergistic combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 2B.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a synergistic combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 3.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination (CBC and THC) of Table 6 A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 6 A marked with *.

Table 6A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combinations of Table 6D marked with *

SUBSTITUTE SHEET (RULE 26) Table 6D ratio of CBC to THC

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 7A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 7A marked with * or **.

Table 7A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combinations of Table 7D marked with * Qj> **

SUBSTITUTE SHEET (RULE 26) Table 7D CBC to THC ratio

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 8 A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 8 A marked with *.

Table 8A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 8D marked with * o **

SUBSTITUTE SHEET (RULE 26) Table 8D CBC to THC ratio

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 9 A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 9 A marked with *.

Table 9A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 9D marked with * o **

SUBSTITUTE SHEET (RULE 26) Table 9D CBC to THC ratio

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 10A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 10A marked with * or ***.

Table 10A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 10D marked with * QJ> ***

SUBSTITUTE SHEET (RULE 26) Table 10D CBC to THC ratio

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 11A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 11A marked with *.

Table 11A delta values calculated according to the Bliss model

In some embodiments, the combination is a synergistic combination for use in reducing viability of ovarian cancer cells and/or in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 1 ID marked with *

SUBSTITUTE SHEET (RULE 26) Table 11D CBC to THC ratio

It should be noted that Table 6A, Table 6D, Table 7A, Table 7D, Table 8A, Table 8D, Table 9A, Table 9D, Table 106A, Table 10D, Table 11A and Table 1 ID are identical to Table 6A, Table 6D, Table 7A, Table 7D, Table 8A, Table 8D, Table 9A, Table 9D, Table 106A, Table 10D, Table 11A and Table 11D provided in the Examples below and are reproduced herein. As detailed in the Examples below and noted herein, synergistic combinations are marked in Table 6A, Table 6D, Table 7A, Table 7D, Table 8A, Table 8D, Table 9A, Table 9D, Table 106A, Table 10D, Table 11A and Table 11D by *, ** or ***. Such combinations in such Tables are synergistic combinations.

In some embodiments, the combination has a synergistic activity in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the combination has a synergistic activity in heating, preventing, or delaying the onset of ovarian cancer in a subject suffering from ovarian cancer or who is predicted to suffer from ovarian cancer or who has been diagnosed to potentially develop ovarian cancer.

In some embodiments, the cancer is lung cancer.

The term lung cancer as referred herein encompasses several different types of cancer that arise in any part of the lung including, inter alia, small-cell lung carcinoma (SCLC), also called oat cell cancer, and non-small-cell lung carcinoma (NSCLC).

Hence, in accordance with some aspects, the present disclosure provides a combination comprising a synergistic amount of CBC and a synergistic amount of THC, for use in reducing viability of lung cancer cells.

In accordance with some other aspects, the present disclosure provides a combination comprising a synergistic amount of CBC and a synergistic amount of THC,

SUBSTITUTE SHEET (RULE 26) for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of at least 1 pg/mL, at times at least 5 pg/mL and THC at a concentration of 1 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 0.5 pg/mL and about 25 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 25 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 5 pg/mL and about 25 pg/mL and THC at a concentration of between about 1 pg/mL and about 20 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 5 pg/mL and about 20 pg/mL and THC at a concentration of between about 1 pg/mL and about 20 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of between about 5 pg/mL and about 10 pg/mL and THC at a concentration of between about 10 pg/mL and about 15 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 10 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 15 pg/mL and 20 pg/mL, (iii) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 5 pg/mL, (iv) CBC at a concentration of about 20 pg/mL and THC at a concentration of between about 1 pg/mL and about 5 pg/mL or (v) any combinations thereof. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC and THC in a concentration ratio of at least one of (i) CBC at a concentration of about 5 pg/mL and THC at a concentration of about 15 pg/mL, (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of about 20 pg/mL, (iii) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 10 pg/mL, (iv) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 15 pg/mL, (v) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 20 pg/mL, (vi) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 5 pg/mL, (vii) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 10 pg/mL, (viii) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 15 pg/mL, (ix) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 20 pg/mL, (x) CBC at a concentration of about 20 pg/mL and THC at a concentration of about 1 pg/mL, (xi) CBC at a concentration of about 20 pg/mL and THC at a concentration of about 5 pg/mL (xii) CBC at a concentration of about 20 pg/mL and THC at a concentration of about 10 pg/mL, (xiii) CBC at a concentration of about 20 pg/mL and THC at a concentration of about 15 pg/mL, (xiv) CBC at a concentration of about 20 pg/mL and THC at a concentration of about 20 pg/mL or (xv) any combination thereof. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of about 5 pg/mL and THC at a concentration of about 15 pg/mL, (ii) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 10 pg/mL or (iii) any combinations thereof. In some embodiments, such a combination is a synergistic combination for use in reducing viability of lung cancer cells and/or for use in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof. In some embodiments, the combination has a synergistic activity against lung cancer.

In some embodiments, the combination has a synergistic activity in reducing cell viability in lung cancer cells.

In some embodiments, the combination comprises between about 10% and about 96% CBD for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a synergistic combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 4.

In some embodiments, the combination is a synergistic combination for use in reducing viability of lung cancer cells and/or in heating, preventing, or delaying the onset of lung cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 12A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of lung cancer cells and/or in heating, preventing, or delaying the onset of lung cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 12A marked with * or ***.

Table 12A delta values calculated according to the Bliss model

SUBSTITUTE SHEET (RULE 26) In some embodiments, the combination is a synergistic combination for use in reducing viability of lung cancer cells and/or in beating, preventing, or delaying the onset of lung cancer in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 12D marked with * or ***.

Table 12D CBC to THC ratio

It should be noted that Table 12A and Table 12D are identical to Table 12A and Table 12D provided in the Examples below and are reproduced herein. As detailed in the Examples below and noted herein, synergistic combinations are marked in Table 12A and Table 12D by *, ** or ***. Such combinations in such Tables are synergistic combinations.

In some embodiments, the combination has a synergistic activity in heating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the combination has a synergistic activity in heating, preventing, or delaying the onset of lung cancer in a subject suffering from lung cancer or who is predicted to suffer from lung cancer or who has been diagnosed to potentially develop lung cancer.

In some embodiments, the cancer is colon cancer.

Colorectal cancer (CRC), also known as bowel cancer, colon cancer, or rectal cancer, as referred herein encompasses several different types of cancer that arise from the colon or rectum (parts of the large intestine), including, inter alia, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, primary colorectal lymphomas and squamous cell carcinomas.

SUBSTITUTE SHEET (RULE 26) Hence, in accordance with some aspects, the present disclosure provides a combination comprising a synergistic amount of CBC and a synergistic amount of THC, for use in reducing viability of colon cancer cells.

In accordance with some other aspects, the present disclosure provides a combination comprising a synergistic amount of CBC and a synergistic amount of THC, for use in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In some embodiments, the combination comprises CBC at a concentration of at least 1 pg/mL and THC at a concentration of 1.1 pg/mL and having a synergistic activity on colon cancer.

In some embodiments, the combination comprises said CBC at a concentration of between about 1 pg/mL and about 15 pg/mL and said THC at a concentration of between about 5 pg/mL and about 20 pg/mL. In some embodiments, such a combination is a synergistic combination for use in reducing viability of colon cancer cells and/or for use in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of between about 5 pg/mL and about 10 pg/mL and THC at a concentration of between about 5 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 15 pg/mL and THC at a concentration of between about 5 pg/mL and 15 pg/mL, (iii) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 10 pg/mL (iv) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 20 pg/mL or (v) any combinations thereof. In some embodiments, such a combination is a synergistic combination for use in reducing viability of colon cancer cells and/or for use in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In some embodiments, the combination comprises at least one of: (i) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 5 pg/mL and about 10 pg/mL, (ii) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 10 pg/mL or (iii) any combinations thereof. In some embodiments, such a combination is a synergistic combination for use in reducing viability of colon cancer cells and/or for use in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof. In some embodiments, the combination has a synergistic activity on colon cancer.

In some embodiments, the combination has a synergistic activity in reducing cell viability in colon cancer cells.

In some embodiments, the combination comprises between about 2% and about 90% CBD for use in reducing viability of ovarian cancer cells and/or for use in beating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In accordance with some embodiments that can be considered as aspects of the present disclosure, it is provided a synergistic combination comprising CBC and THC for use in reducing cancer cell viability and/or treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one active combination or the combination having a CBC to THC ratio as in at least one active combination as provided in Table 5.

In some embodiments, the combination is a synergistic combination for use in reducing viability of colon cancer cells and/or in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 13A having a delta value >1.

In some embodiments, the combination is a synergistic combination for use in reducing viability of colon cancer cells and/or in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof, wherein the synergistic combination is or comprises at least one combination of Table 13A marked with * or ***.

Table 13A delta values calculated according to the Bliss model

SUBSTITUTE SHEET (RULE 26) In some embodiments, the combination is a synergistic combination for use in reducing viability of colon cancer cells and/or in treating, preventing, or delaying the onset of colon in a subject in need thereof, wherein the synergistic combination having a CBC to THC ratio as in any one of the combination of Table 13D marked with * or ***.

Table 13D CBC to THC ratio

It should be noted that Table 13A and Table 13D are identical to Table 13A and Table 13D provided in the Examples below and are reproduced herein. As detailed in the Examples below and noted herein, synergistic combinations are marked in Table 13A and Table 13D by *, ** or ***. Such combinations in such Tables are synergistic combinations.

In some embodiments, the combination has a synergistic activity in beating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In some embodiments, the combination has a synergistic activity in heating, preventing, or delaying the onset of colon cancer in a subject suffering from colon cancer or who is predicted to suffer from colon cancer or who has been diagnosed to potentially develop colon cancer.

As described herein, the present disclosure refers to combinations of CBC and THC. Combinations of CBC and THC encompasses combinations in which each one of CBC and THC is in a different dosage form as well as combinations in which CBC and THC are present together in a single dosage forms.

In some embodiments, CBC is present in first composition and THC is present in a second, different composition. In other words, CBC and THC are not in the same composition.

SUBSTITUTE SHEET (RULE 26) In some embodiments, CBC and THC are present in the same composition.

Thus, the present disclosure also provides a pharmaceutical composition comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The present disclosure also provides a pharmaceutical composition comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In some embodiments, the concentration of CBC in the pharmaceutical composition is at least about 0.5% (w/w), at times at least about 1%, at times at least about 2%, at times at least about 3%, at times at least about 4%, at times at least about 5%, at times at least about 6%, at times at least about 10%, at times at least about 15%, at times at least about 20%, at times at least about 25%, at times at least about 30%, at times at least about 35%, at times at least about 40%, at times at least about 45%, at times at least about 50%, at times at least about 55%, at times at least about 60%, at times at least about 65%, at times at least about 70%, at times at least about 80%, at times at least about 85%, at times at least about 90%, at times at least about 95%. Any value which is between any one of the above values and/or any range between one or more of the above values is within the scope of the present disclosure.

In some embodiments, the concentration of CBC in the pharmaceutical composition is between about 0.5% and about 97%.

In some embodiments, the concentration of THC in the pharmaceutical composition is at least about 0.5% (w/w), at times at least about 1%, at times at least about 2%, at times at least about 3%, at times at least about 4%, at times at least about 5%, at times at least about 6%, at times at least about 10%, at times at least about 15%, at times at least about 20%, at times at least about 25%, at times at least about 30%, at times at least about 35%, at times at least about 40%, at times at least about 45%, at times at least about 50%, at times at least about 55%, at times at least about 60%, at times at least about 65%, at times at least about 70%, at times at least about 80%, at times at least about 85%, at times at least about 90%, at times at least about 95%. Any value which is between any one of the above values and/or any range between one or more of the above values is within the scope of the present disclosure.

In some embodiments, the concentration of THC in the pharmaceutical composition is between about 0.5% and about 97%.

In some embodiments, the pharmaceutical composition comprises at least one pharmaceutically acceptable excipient or carrier.

As used herein “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated.

The pharmaceutical composition of the invention can be administered and dosed by the methods of the invention, in accordance with good medical practice. The combinations, compositions used in the methods and kits of the invention, described herein after, may be adapted for administration by systemic, parenteral, intraperitoneal, transdermal, oral (including buccal or sublingual), rectal, topical (including buccal or sublingual), vaginal, intranasal and any other appropriate routes. Such formulations may be prepared by any method known in the art of pharmacy, for example by bringing into association the active ingredient with the carrier(s) or excipient(s).

The phrases "systemic administration," "administered systemically," "peripheral administration" and "administered peripherally" as used herein mean the administration of a compound, drug or other material other than directly into the central blood system, such that it enters the patient's system and, thus, is subject to metabolism and other like processes. The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion. Regardless of the route of administration selected, CBC and/or THC may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically-acceptable dosage forms by conventional methods known to those of skill in the art.

The pharmaceutical forms suitable for injection use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

Pharmaceutical compositions used to treat subjects in need thereof according to the invention generally comprise a buffering agent, an agent who adjusts the osmolarity thereof, and optionally, one or more pharmaceutically acceptable carriers, excipients and/or additives as known in the art. Supplementary active ingredients can also be incorporated into the compositions. The carrier can be solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

Local administration to the area in need of treatment may be achieved by, for example, local infusion during surgery, topical application, direct injection into the specific organ, etc.

Compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, lozenges (including liquid-filled), chews, multi- and nano-particulates, gels, solid solution, liposome, films, ovules, sprays or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations may also include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents. In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations and concentrations ratios according to at least one of (i) Table 1, (ii) Table 2 A, (iii) Table 2B, (iv) Table 3, (v) Table 4, (vi) Table 5 or (vii) any combinations thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations according to any one of Table 6A, Table 7A, Table 8A, Table 9A, Table 10A, Table 11A or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC having least one ratio according to any one of Table 6D, Table 7 D, Table 8 D, Table 9 D, Table 10 D, Table 1 ID or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations according to any one of Table 6A, Table 7A, Table 8A, or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC having least one ratio according to any one of Table 6D, Table 7D, Table 8 D, or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations according to any one of Table 9 A, Table 10A, Table 11A or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC having least one ratio according to any one of Table 9D, Table 10D, Table 11D or any combination thereof.

In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations according to any one of Table 12 A.

In some embodiments, the pharmaceutical composition comprising CBC and THC having least one ratio according to any one of Table 12D.

In some embodiments, the pharmaceutical composition comprising CBC and THC in concentrations according to any one of Table 13 A.

In some embodiments, the pharmaceutical composition comprising CBC and THC having least one ratio according to any one of Table 13D. In some embodiments, the pharmaceutical composition has a synergistic activity on ovarian cancer.

In some embodiments, the pharmaceutical composition has a synergistic activity in reducing cell viability in ovarian cancer cells.

In some embodiments, the pharmaceutical composition has a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises CBC at a concentration of at least 0.5 pg/mL and THC at a concentration of at least 0.5 pg/mL and having a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations of any one of Table 6A, Table 7A, Table 8A or any combination of Table 6A, Table 7A and Table 8A and having a synergistic activity in reducing viability of ovarian cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations of any one of Table 9 A, Table 10 A, Table 11A or any combination of Table 9 A, Table 10A and Table 11A and having a synergistic activity in reducing viability of ovarian cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations having a CBC to THC ratio of any one of Table 6D, Table 7 D, Table 8D, Table 9D, Table 10D, Table 11D or any combination of Table 6D, Table 7D, Table 8D, Table 9D, Table 10D and Table 1 ID and having a synergistic activity in reducing viability of ovarian cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations having a CBC to THC ratio of any one of Table 6D, Table 7D, Table 8D, or any combination of Table 6D, Table 7D, Table 8D and having a synergistic activity in reducing viability of ovarian cancer cells and/or having a synergistic activity in beating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises synergistic combinations having a CBC to THC ratio of any one of Table 9D, Table 10D, Table 1 ID or any combination of Table 9D, Table 10D and Table 11D and having a synergistic activity in reducing viability of ovarian cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition has a synergistic activity on lung cancer.

In some embodiments, the pharmaceutical composition comprises CBC at a concentration of at least 1 pg/mL and THC at a concentration of 1.1 pg/mL and having a synergistic activity in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations of any one of Table 12A and having a synergistic activity in reducing viability of lung cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations having a CBC to THC ratio of any one of Table 12D and having a synergistic activity in reducing viability of lung cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of lung cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition has a synergistic activity on colon cancer.

In some embodiments, the pharmaceutical composition comprises CBC at a concentration of at least 0.5 pg/mL and THC at a concentration of 0.5 pg/mL and having a synergistic activity in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

In some embodiments, the pharmaceutical composition comprises synergistic combinations of any one of Table 13A and having a synergistic activity in reducing viability of colon cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof. In some embodiments, the pharmaceutical composition comprises synergistic combinations having a CBC to THC ratio of any one of Table 13D and having a synergistic activity in reducing viability of colon cancer cells and/or having a synergistic activity in treating, preventing, or delaying the onset of colon cancer in a subject in need thereof.

According to a further aspect of the present disclosure it is provided a method for reducing cell viability, the method comprising contacting the cell with a combination comprising CBC and THC, wherein the CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and the THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

The term “contacting” means to bring, put, incubates or mix together. As such, a first item is contacted with a second item when the two items are brought or put together, e.g., by touching them to each other or combining them. In the context of the present invention, the term "contacting" includes all measures or steps which allow interaction between CBC, THC and a cell, specifically a cancer cell. In some examples, the cell is an ovarian cancer cell, a lung cancer cell, a colon cancer cell or any combination thereof.

In some embodiments, the method of the present disclosure comprises inducing cell cycle arrest and/or reducing migration of cancer cells.

The methods of the invention are capable of at least one of (i) reducing cell viability, (ii) inducing cell cycle arrest, (iii) reducing migration of cancer cells or (iv) any combinations thereof.

The methods of the invention can be in vitro methods and/or in vivo methods.

In some embodiments, the method is an in vitro method.

In some other embodiments, the method is an in in vivo method.

As appreciated, any one of (i) reducing cell viability, (ii) inducing cell cycle arrest, (iii) reducing migration of cancer cells or (iv) any combinations thereof is effective in cancer therapy.

According to a further aspect of the present disclosure it is provided a method of treating, preventing, or delaying the onset of cancer in a subject in need thereof, the method comprising administering to the subject a combination comprising CBC and THC, wherein the CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and the THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In accordance with some embodiments, the subject in need thereof is a subject suffering from cancer or who is predicted to suffer from cancer or who is predicted to suffer from cancer or who has been diagnosed to potentially develop cancer.

In accordance with the present invention, the subject to be treated with a combination, as disclosed herein, is one which has been found or determined to possibly benefit from the treatment with the combination and specifically the synergistic combination, and for whom a therapeutic protocol has been tailored, specifying amounts of the CBC and THC, such that the amount of one component is adjusted to or determined based on the amount of the other component or to any one other protocol parameter which may, inter alia, depend on subject health and personal factors as well as on time of administration, sequence and administration regimen.

As noted herein above, the combination of CBC and THC can be present in a single composition or each one of CBC and THC is present in a different composition.

Hence, it should be understood that methods of the invention making use of administration of CBC with THC in "combination" (at times denoted "together"') refers to a multiple’s treatment schedules.

In accordance with some embodiments, combination denotes a regimen involving administration of at least CBC and THC by the methods of the invention in a single treatment cycle. As may be appreciated, administration of the combination requires an assessment of various parameters, inter alia, treatment schedule, predetermined ratio of the CBC and THC (dosing), number of treatment cycles, and others. Assessment of the combination may be carried out before onset of treatment.

The CBC and THC described herein can be administered and dosed by the methods of the invention, in accordance with good medical practice, such as systemically, for example by parenteral, e.g. intravenous, intraperitoneal or intramuscular injection.

In some embodiments, the administration of CBC and THC by the methods of the invention may be by the same or different modes of administration and at the same or different frequency, i.e. at the same or different time points. In accordance with some embodiments, administration of the CBC and the THC by the methods of the invention is at different dosage forms. In other words, CBC and the THC are not present in the same composition.

As detailed above, various modes of administration are known in the art, including, but are not limiting to, topical administration, enteral administration, parental administration. For example by injection (e.g., using a subcutaneous, intramuscular, intravenous, intraperitoneal, or intradermal injection), infusion (e.g. intraperitoneal), transdermal, transmucosal, intranasal administration, by inhalation or sublingually.

In some embodiments, CBC and the THC are administered by the methods of the invention in different routes.

In some embodiments, each of CBC and the THC are administered by the methods of the invention separately, i.e. not within the same composition/dosage form.

In some other embodiments, the CBC and the THC are administered by the methods of the invention simultaneously.

As noted above and in accordance with some embodiments, the CBC and THC are present in the same composition.

In some embodiments, the method is of treating, preventing, or delaying the onset of ovarian cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In some embodiments, the method is of treating, preventing, or delaying the onset of lung cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In some embodiments, the method is of treating, preventing, or delaying the onset of colon cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In accordance with some further aspects, the present disclosure provided a kit comprising CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

In some embodiments, the kit comprises CBC and THC each in separate reservoirs.

In some embodiments, the different reservoirs are different syringes or different formulation containers comprising the actives in solid or liquid or solution forms.

In some embodiments, the kit comprise a container means for containing separate compositions; such as a divided bottle or a divided foil packet.

In some embodiments, the kit includes directions for the administration of the separate components. It should be understood that such a kit is advantageous when the separate components are preferably administered in different dosage forms, are administered at different dosage intervals, or when titration of the individual components of the combination is desired by the prescribing physician.

It should be appreciated that each of the multiple components of the kit may be administered simultaneously.

Alternatively, each of said multiple dosage forms may be administered sequentially in either order.

Further, the kits described herein can include a composition as described, or in separate multiple dosage unit forms, as an already prepared dosage form ready for administration or, alternatively, can include the composition as described as a solid pharmaceutical composition that can be reconstituted with a solvent to provide a liquid oral dosage form.

When the kit includes a solid pharmaceutical composition that can be reconstituted with a solvent to provide a liquid dosage form (e.g., for oral administration), the kit may optionally include a reconstituting solvent.

In some embodiments, the kit comprises CBC and THC in the same composition. In some embodiments, the kit comprising instructions for using CBC and THC in a combination for treating, preventing, or delaying the onset of cancer in a subject in need thereof.

In accordance with some further aspects, the present disclosure provides use of CBC and THC in the preparation of a pharmaceutical composition, the pharmaceutical composition comprising CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

As noted herein, the CBC and the THC may each be administrated to a subject in need thereof. Administration of each one of CBC and THC is in an effective amount or a therapeutically effective dose.

The term "effective amount" or “therapeutically effective dose” as used herein is that determined by such considerations as are known to the man of skill in the art. The amount must be sufficient to treat, prevent or delay the onset of cancer and specifically, a cancer selected from ovarian cancer, lung cancer and colon cancer.

As known, the "effective amount" for purposes herein may be determined by such considerations as known in the art. The amount must be effective to achieve the desired therapeutic effect, depending, inter alia, on the type and severity of the disease to be treated and the treatment regime. As generally known, the effective amount depends on a variety of factors including for example the distribution profile within the body, a variety of pharmacological parameters such as half-life in the body, on undesired side effects, if any, on factors such as age and gender, and others. In the context of the present disclosure and as described herein, the effective amount of the cannabis plant extract and the effective amount of GA are selected such that the combination has the desired therapeutic effect.

In specific embodiments, the amount must be sufficient to prevent or inhibit relapse of a disease, specifically cancer as described herein.

The effective amount and/or therapeutically effective dose of CBC and THC in accordance with some embodiments is a synergistic amount.

It should be noted that determination of an effective amount, therapeutically effective amount or synergistic amount or any of these amounts is well within the skills of a person skilled in the art as described herein and specifically in view of the disclosure herein, including the Examples provided below. Any combination or pharmaceutically composition used in the invention, for example in the methods of the invention, the effective amount, therapeutically effective amount or synergistic amount or dose can be estimated from in vitro and cell culture assays described herein. For example, a dose can be formulated in animal models to achieve a required concentration or titer. Such information can be used to accurately determine useful doses in humans.

As appreciated, doses determined in the animal model such as mouse model can be converted for the treatment other species such as human and other animals diagnosed with the disease, using conversion Tables known to the skilled in the art.

Hence, as appreciated by those skilled in the art, the in vitro data shown herein from cell culture assays can be used with animal studies to formulate a range of dosage for use in human.

The term “treatment or prevention” as used herein, refers to the complete range of therapeutically positive effects of administrating to a subject including inhibition, reduction of, alleviation of, and relief from, a cancerous condition and illness, cancer symptoms or undesired side effects or cancer related disorders. More specifically, treatment or prevention of relapse or recurrence of the disease in response to a treatment with a non-effective, or deleterious therapeutic agent, includes the prevention or postponement of development of the disease, prevention or postponement of development of symptoms and/or a reduction in the severity of such symptoms that will or are expected to develop. These further include ameliorating existing symptoms, preventing- additional symptoms and ameliorating or preventing the underlying metabolic causes of symptoms. It should be appreciated that the terms "inhibition", "moderation", “reduction” or "attenuation" as referred to herein, relate to the retardation, restraining or reduction of a process by any one of about 1% to 99.9%, specifically, about 1% to about 5%, about 5% to 10%, about 10% to 15%, about 15% to 20%, about 20% to 25%, about 25% to 30%, about 30% to 35%, about 35% to 40%, about 40% to 45%, about 45% to 50%, about 50% to 55%, about 55% to 60%, about 60% to 65%, about 65% to 70%, about 75% to 80%, about 80% to 85% about 85% to 90%, about 90% to 95%, about 95% to 99%, or about 99% to 99.9%. It is understood that the interchangeably used terms "associated", “linked” and "related", when referring to pathologies herein, mean diseases, disorders, conditions, or any pathologies which at least one of: share causalities, co-exist at a higher than coincidental frequency, or where at least one disease, disorder condition or pathology causes the second disease, disorder, condition or pathology.

The present invention relates to the treatment of subjects, or patients, in need thereof. By “patient” or “subject in need” it is meant any organism who may be affected by the above-mentioned conditions, and to whom the treatment methods herein described are desired, including humans, domestic and non-domestic mammals such as canine and feline subjects, bovine, simian, equine and murine subjects, rodents, domestic birds, aquaculture, fish and exotic aquarium fish. It should be appreciated that the treated subject may be also any reptile or zoo animal. More specifically, the methods and compositions of the invention are intended for mammals. By “mammalian subject” is meant any mammal for which the proposed therapy is desired, including human, equine, canine, and feline subjects, most specifically humans. It should be noted that specifically in cases of non-human subjects, the method of the invention may be performed using administration via injection, drinking water, feed, spraying, oral gavage and directly into the digestive tract of subjects in need thereof. It should be further noted that particularly in case of human subject, administering of the compositions of the invention to the patient includes both self-administration and administration to the patient by another person.

The term "about" as used herein indicates values that may deviate up to 1%, more specifically 5%, more specifically 10%, more specifically 15%, and in some cases up to 20% higher or lower than the value referred to, the deviation range including integer values, and, if applicable, non-integer values as well, constituting a continuous range. As used herein the term "about" refers to ± 10 %.

It should be noted that various embodiments of this invention may be presented in a range format. The description of a range should be considered to have specifically disclosed all the possible sub ranges as well as individual numerical values within that range inclusive the end points values. For example, description of a range such as from 1 to 6 or between 1 and 6 should be considered to have specifically disclosed sub ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. It should be noted that the term “at least one” refers to one or more of the following and encompasses any one of 1, 2, 3, or more of the specified information.

It is to be understood that the terminology used herein is used for the purpose of describing particular embodiments only and not intended to be limiting since the scope of the present invention will be limited only by the appended claims and equivalents thereof.

Throughout this specification and the Examples and claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

It must be noted that, as used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise.

The following examples are representative of techniques employed by the inventors in carrying out aspects of the present invention. It should be appreciated that while these techniques are exemplary of preferred embodiments for the practice of the invention, those of skill in the art, in light of the present disclosure, will recognize that numerous modifications can be made without departing from the spirit and intended scope of the invention.

It should be noted that the various embodiments and examples detailed herein in connection with various aspects of the invention may be applicable to one or more aspects disclosed herein. It should be further noted that any embodiment described herein, for example, related to components of the microbial consortium, may be applied separately or in various combinations. Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples. The phrases “in another embodiment” or any refence made to embodiment as used herein do not necessarily refer to different embodiment, although it may. Thus, various embodiments of the invention can be combined (from the same or from different aspects) without departing from the scope of the invention. Table 1 Concentrations and ratios of CBC and THC having activity against multiple cancer cells

Table 1 (Cont.)

Table 2A Concentrations and ratios of CBC and THC with activity in ovarian cancer.

Table 2B Concentrations and ratios of CBC and THC having activity in ovarian cancer

Table 3 Concentrations and ratios of CBC and THC having activity in ovarian cancer

Table 4 Concentrations and ratios of CBC and THC having activity in lung cancer.

Table 5 Concentrations and ratios of CBC and THC having activity in colon cancer.

NON-LIMITING EXAMPLES

Material and Methods

Standard/material preparation and use

The cannabinoid standards (at a concentration of 1 mg/mL in methanol) used in this study included delta-9 -tetrahydrocannabidiol (A-9 THC, Restek catalog no. 34067) and cannabichromene (CBC, Restek catalog no. 34092).

Cell culture

Cell lines: HTB75 (ATCC, CAOV-3; Adenocarcinoma), A549 (ATCC, CCL-185; Epithelial Lung Carcinoma), were cultured in Dulbecco's Modified Eagle Medium (DMEM) medium (01-055-1A, Biological Industries, Israel) supplemented with 10% fetal bovine serum (FBS) (04-127-1 A, Biological Industries, Israel), HTB 161 (ATCC, OVCAR-3; Adenocarcinoma) was cultured in RPMI medium (01-100-1A, Biological Industries, Israel), supplemented with 20% FBS. HCT116 (ATCC, CCL-247; Epithelial Colorectal Carcinoma) was cultured in McCoy's 5 A Medium (01 -075-1 A, Biological Industries, Israel) supplemented with 10% fetal bovine serum (FBS) (04-127-1A, Biological Industries, Israel). All media were supplemented with 1% Pen-Strep, 1% L- Glutamine and 0.02% plasmocin. Cells were incubated in 37 °C in a humidified atmosphere, in environmental containing 5% CO2-95% air.

Cell viability assay

Cells were seeded into 96- well plates at density of 2 x 10⁴ per well (100 pL/well) and were incubated at 37 °C overnight to allowed attachment. The following day, cells were treated with cannabinoid standards in volume of 100 pL/well at different concentrations. Solvents used as a vehicle control. Treated cells were incubated for 48 h at 37 °C. Subsequently, XTT reagents (2, 3, -bis (2-methoxy- 4-nitro- 5-sulfophenyl)- 5- [(phenylamino)- carbonyl]-2H- tetrazolium inner salt) (20-300-1000; Biological Industries, Israel) were added to the cells for 2 h at 37 °C in a humidified 5% CC>2-95% air atmosphere. Absorbance was recorded by a Synergy Hl hybrid reader photometer (BioTek) at 490 nm with 650 nm of reference wavelength. Cell viability was estimated from the equation: (A490 — A650) of treatment

% Cell Viability = 100 x

(A490 — A650) of solvent contro

A490 and A650 are the absorbencies of the XTT colorimetric reaction. Absorbance of medium alone (blank) was subtracted from the readings. For dose response assays, data points were connected by non-linear regression lines of the sigmoidal dose-response relation. GraphPad Prism version 6.1 (https://www.graphpad.com/scientific- software/prism/, GraphPad Software Inc., San Diego, USA) was employed to produce dose-response curves and determination of IC50 values.

Analysis of combined drug effects

Analysis of combined drug effect was determined from XTT results. Drug synergy was determined by two synergy models.

1. The Bliss independence drug interaction model: defined by the following equation: Exy = Ex + Ey - (ExEy), where (Exy) is the additive effect of the drugs x and y as predicted (calculated) by their individual effects (Ex and Ey). The synergy expressed by the delta of EXy and the observed value. The delta values differ on a scale between 0 [no synergy] to 1 [high synergy].

2. The Loewe additivity model: Assuming that two compounds have the same mechanism of action and therefore one compound can be a substitute for another. Here we compare the effect (% viability) of the treatment combined with two active components (dl and d2) to each one of them alone, xl and x2 are the doses at which each drug alone produces the response, respectively, fl (x) and 2(v) are the dose-response functions for dl and d2, respectively, While the function is the required concentration of each drug to achieve the same response when given alone. In case of no interaction between the compounds, the dose combinations will have the same response and will follow the equation: xl/fl + xl/fl = 1. Values below 1 will represent synergism and above 1 antagonism. (Lederer, S., Dijkstra, T.M. and Hcskes, T., 2018. Additive dose response models: explicit formulation and the loewe additivity consistency condition. Frontiers in pharmacology. 9. p.31.)

Statistical Analysis: Data were processed using the JMP statistical package (https://www.jmp.com/en_us/home.html, SAS Inc, NC, USA); mean of replicates compared between two groups were made using the Student’s t-test. Mean of replicates compared between more than 2 groups were made with analysis of variance (ANOVA) followed by Tukey-Kramer's honest significant difference (HSD) test as post hoc. P values < 0.05 were considered significant. Levels with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Results

Example 1: Combinations ofTHC and CBC have synergistic cytotoxic activity against multiple carcinoma cells

The activity of THC or CBC on different epithelial cancer cell lines was determined. These include ovarian adenocarcinoma (HTB75 and HTB161), lung adenocarcinoma (A549) and colorectal carcinoma (HCT116).

ICso of the compounds are presented in Figs. 1A-1H Following, activity of different concentrations of THC and CBC were examined on multiple cancer cells.

In HTB161 cells, IC50 value for THC was 21.63 pg/mL (Fig. 1A) and for CBC was 20.15 pg/mL (Fig. IB).

In HTB75 cells, IC50 value for THC was 12.33 pg/mL (Fig. 1C) and for CBC was 9.83 pg/mL (Fig. ID).

In A549 cells, IC50 value for THC was 24.27 pg/mL (Fig. IE) and for CBC was 22.45 pg/mL (Fig. IF).

In HCT116 cells, IC50 value for THC was 20.73 pg/mL (Fig. 1G) and for CBC was 19.38 pg/mL (Fig. 1H).

Example 2: Ovarian adenocarcinoma cell lines

A significant synergy was found for combinations of THC and CBC on ovarian cancer (OC) cells, HTB75, at 24h, 48h and 72h.

Fig. 2A shows synergy for combinations of THC and CBC at 24 hours, Fig. 2B shows synergy for combinations ofTHC and CBC at 48 hours and Fig.2C shows synergy for combinations of THC and CBC at 72 hours in HTB75 cells. Tables 6A-6C demonstrate synergistic activity on cell viability using combination of CBC and THC in various concentrations in HTB75 cells after 24 hours, Table 6A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 6B shows standard error values calculated for n=3, Table 6C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 6D and 6E show calculations of the CBC and THC ratios, Table 6D CBC/THC and Table 6E THC/CBC.

Table 6 A and Table 6D in this example are identical to Table 6 A and Table 6D provided in the detailed description part.

Combinations of CBC and THC concentrations marked by ** that were found to be synergistic using Loewe model and in* are concentrations that were found to be synergistic using Bliss model.

Tables 6A to 6E: synergistic activity against HTB75 cell viability after 24 hours.

Table 6A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 24 hours.

Table 6B Standard error calculations n=3

Table 6C synergism statistics

SUBSTITUTE SHEET (RULE 26) Table 6D ratio of CBC to THC

Table 6E ratio of THC to CBC

SUBSTITUTE SHEET (RULE 26) Tables 7A - 7C demonstrate synergistic activity on cell viability using CBC and THC in various concentrations of CBC and THC in HTB75 cells after 48 hours, Table 7A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 7B shows standard error values calculated for n=3, Table 7C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 7D and 7E show calculations of the CBC and THC ratios, Table 7D CBC/THC and Table 7E THC/CBC. Combinations of concentrations marked by ** that were found to be synergistic using Loewe model and in* are concentrations that were found to be synergistic using Bliss model.

Table 7A and Table 7D in this example are identical to Table 7A and Table 7D provided in the detailed description part.

Tables 7A to 7E: synergistic activity against HTB75 cell viability after 48 hours.

Table 7A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 48 hours,

Table 7B Standard error calculations

SUBSTITUTE SHEET (RULE 26) Table 7C synergism statistics

Table 7D CBC to THC ratio

Table 7E THC to CBC ratio

SUBSTITUTE SHEET (RULE 26) Tables 8A - 8C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in HTB75 cells after 72 hours, Table 8A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 8B shows standard error values calculated for n=3, Table 8C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 8D and 8E show calculations of the CBC and THC ratios, Table 8D CBC/THC and Table 8E THC/CBC.

Table 8 A and Table 8D in this example are identical to Table 8 A and Table 8D provided in the detailed description part.

Combinations of concentrations marked by ** that were found to be synergistic using Loewe model and in* are concentrations that were found to be synergistic using Bliss model.

Tables 8A to 8E: synergistic activity against HTB75 cell viability after 72 hours.

Table 8A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 72 hours

Table 8B Standard error calculations n=3

SUBSTITUTE SHEET (RULE 26) Table 8C synergism statistics

Table 8D CBC to THC ratio

SUBSTITUTE SHEET (RULE 26) Table 8E THC to CBC ratio

As shown in Figs. 2A-2C and Tables 6A-6E, 7A-7E and 8A-8E, synergy was observed in a variety of combinations of THC and CBC in all tested times (24h, 48h and 72h). when the tested time was increased to 72 hours, synergy was observed at lower concentration of THC and CBC, suggesting that the activity of THC and CBC is time deepened.

As further shown in Tables 7A and 7C, synergy calculated using the Loewe model, was observed in a CBC concentration of 0.5 pg/mL and THC concentration of 5 pg/mL. The Loewe value for this combination was 0.52.

SUBSTITUTE SHEET (RULE 26) Example 3: Ovarian adenocarcinoma cell lines

A significant synergy was found for combinations of THC and CBC on ovarian cancer (OC) cells, HTB161, at 24h, 48h and 72h.

Fig. 3A shows synergy for combinations of THC and CBC at 24 hours, Fig. 3B shows synergy for combinations of THC and CBC at 48 hours and Fig. 3C shows synergy for combinations of THC and CBC at 72 hours in HTB161 cells.

Tables 9A-9C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in HTB161 cells after 24 hours such that synergism is considered when the delta value is>0, Table 9A shows delta values calculated according to the Bliss model, Table 9B shows standard error values calculated for n=3, Table 9C shows synergy statistics._Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 9D and 9E show calculations of the CBC and THC ratios, Table 9D CBC/THC and Table 9E THC/CBC.

Table 9 A and Table 9D in this example are identical to Table 9 A and Table 9D provided in the detailed description part.

Tables 9A to 9E: synergistic activity against HTB161 cell viability after 24 hours.

Table 9A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 24 hours

Table 9B Standard error calculations n=3

Table 9C synergism statistics

SUBSTITUTE SHEET (RULE 26) Table 9D CBC/THC

Table 9E THC/CBC

SUBSTITUTE SHEET (RULE 26) Tables 10A-10C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in HTB161 cells after 48 hours, Table 10A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 10B shows standard error values calculated for n=3, Table 10C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 10D and 10E show calculations of the CBC and THC ratios, Table 10D CBC/THC and Table 10E THC/CBC.

Table 10A and Table 10D in this example are identical to Table 10A and Table 10D provided in the detailed description part.

Combinations of concentrations marked by *** that were found to be synergistic using both Bliss and Loewe models and in* are concentrations that were found to be synergistic using Bliss model.

Tables 10A to 10E: synergistic activity against HTB161 cell viability after 48 hours.

Table 10A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 48 hours.

Table 10B Standard error calculations n=3

SUBSTITUTE SHEET (RULE 26) Table IOC synergism statistics

Table 1OD CBC/THC

SUBSTITUTE SHEET (RULE 26) Table 10E THC/CBC

Tables 11A-11C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in HTB161 cells after 72 hours, Table 11A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 11B shows standard error values calculated for n=3, Table 11C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 11D and HE show calculations of the CBC and THC ratios, Table 11D CBC/THC and Table HE THC/CBC.

Table HA and Table HD in this example are identical to Table HA and Table 1 ID provided in the detailed description part.

Combinations of concentrations marked by ** were found to be synergistic using Loewe model and in * are concentrations that were found to be synergistic using Bliss model.

SUBSTITUTE SHEET (RULE 26) Tables 11A to HE: synergistic activity against HTB161 cell viability after 72 hours.

Table HA delta values calculated according to the Bliss model for various concentrations of CBC and THC after 72 hours.

Table 11B Standard error calculations n=3

Table 11C synergism statistics

SUBSTITUTE SHEET (RULE 26) Table 11D CBC/THC

Table HE THC/CBC

SUBSTITUTE SHEET (RULE 26) As can be seen in Figs. 3A-3C and Tables 9A-9E, lOA-lOE and 11A-11E, synergy was observed in a variety of combinations of THC and CBC in all tested times (24h, 48h and 72h). when the tested time was increased to 72 hours, synergy was observed at lower concentration of THC and CBC, suggesting that the activity of THC and CBC is time dependent.

As further shown in Tables 10A and IOC, synergy calculated using the Loewe model, was observed in multiple combinations of CBC concentrations and THC concentrations as follows: CBC concentration of 0.5pg/mL and THC concentration of 15pg/mL - Loewe value for this combination was 0.90; CBC concentration of 0.5pg/mL and THC concentration of 20pg/mL- Loewe value for this combination was 0.77; CBC concentration of 5pg/mL and THC concentration of lOpg/mL- Loewe value for this combination was 0.25; CBC concentration of 5pg/mL and THC concentration of 15pg/mL- Loewe value for this combination was 0.86; CBC concentration of lOpg/mL and THC concentration of lOpg/mL- Loewe value for this combination was 0.57.

Example 4: Lung adenocarcinoma cell line

A significant synergy was found for combinations of THC and CBC on Lung adenocarcinoma cell line (A549) at 48h (Fig. 4). In various concentrations, synergy was also found using the Loewe index.

Tables 12A -12C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in A549 cells after 48 hours, Table 12A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 12B shows standard error values calculated for n=3, Table 12C shows synergy statistics. Values with different letters are significantly different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 12D and 12E show calculations of the CBC and THC ratios, Table 12D CBC/THC and Table 12E THC/CBC.

Table 12A and Table 12D in this example are identical to Table 12A and Table 12D provided in the detailed description part.

Combinations of concentrations marked by *** were found to be synergistic using both Bliss model and Loewe model and in *are concentrations that were found to be synergistic using Bliss model.

Tables 12A to 12E: synergistic activity against A549 cell viability after 48 hours.

Table 12A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 48 hours.

SUBSTITUTE SHEET (RULE 26) Table 12B: Standard error calculations n=3

Table 12C synergism statistics

Table 12D CBC/THC

SUBSTITUTE SHEET (RULE 26)

Table 12E THC/CBC

As can be seen in Fig. 4 and Tables 12A-12E, synergy was observed in a variety of combinations of THC and CBC.

As further shown in Tables 12A and 12C, synergy calculated using the Loewe model, was observed in several combinations of CBC concentrations and THC concentrations as follows: CBC concentration of 5|ig/mL and THC concentration of 10|ig/mL - Loewe value for this combination was 0.67; CBC concentration of 10|ig/mL and THC concentration of 10|ig/mL- Loewe value for this combination was 0.85.

Example 5: Colorectal carcinoma cell line

A significant synergy was found for combinations of THC and CBC on colorectal carcinoma cell line (HCT116) at 48h (Fig. 5). In Table 3 concentrations that were found to be synergistic in Bliss and/or Loewe additivity and combination index.

Tables 13A - 13C demonstrate synergistic activity on cell viability using CBC an THC in various concentrations of CBC and THC in HCT116cells after 48 hours, Table 13A shows delta values calculated according to the Bliss model such that synergism is considered when the delta value is > 0, Table 13B shows standard error values calculated for n=3, Table 13C shows synergy statistics. Values with different letters are significantly

SUBSTITUTE SHEET (RULE 26) different from all combinations of pairs according to the Tukey-Kramer honest significant difference (HSD; P < 0.05).

Tables 13D and 13E show calculations of the CBC and THC ratios, Table 13D CBC/THC and Table 13E THC/CBC.

Table 13A and Table 13D in this example are identical to Table 13A and Table 13D provided in the detailed description part.

Combinations of concentrations marked by *** were found to be synergistic using both Bliss model and Loewe model and in * are concentrations that were found to be synergistic using Bliss model.

Tables 13A to 13E: synergistic activity against HCT116 cell viability after 48 hours.

Table 13A delta values calculated according to the Bliss model for various concentrations of CBC and THC after 48 hours.

Table 13B Standard error calculations n=3

Table 13C synergism statistics

SUBSTITUTE SHEET (RULE 26) Table 13D CBC/THC.

Table 13E THC/CBC

As can be seen in Fig. 5 and Tables 13A-13E, synergy was observed in a variety of combinations of THC and CBC.

As further shown in Tables 13A and 13C, synergy calculated using the Loewe model, was observed in several combinations of CBC concentrations and THC concentrations as follows: CBC concentration of Igg/mL and THC concentration of lOgg/mL - Loewe value for this combination was 0.23; CBC concentration of 5gg/mL and THC concentration of 5gg/mL- Loewe value for this combination was 0.61; CBC concentration of 5gg/mL and THC concentration of lOgg/mL- Loewe value for this combination was 0.67.

SUBSTITUTE SHEET (RULE 26)

Claims

CLAIMS:

1. A combination comprising CBC and THC for use in reducing cell viability, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

2. A combination comprising CBC and THC for use in treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

3. The combination for use of claim 1 or 2, wherein said CBC and said THC are each present in the combination in a concentration capable of providing a synergistic effect.

4. The combination for use of any one of claims 1 to 3, comprises CBC between about 0.5% and about 97%.

5. The combination for use of any one of claims 1 to 3, comprises said CBC and said THC in concentrations and concentrations ratios according to any one of Table 1 ± 10 %, Table 2A ± 10 %, Table 2B ± 10 %, Table 3 + 10 %, Table 4 ± 10 % or Table 5 + 10 %.

6. The combination for use of any one of claims 1 to 5, wherein the cancer is selected from the group consisting of ovarian cancer, lung cancer, colon cancer.

7. The combination for use of any one of claims 1 to 6, wherein said composition has a synergistic activity on ovarian cancer.

8. The combination for use of claim 7, comprises said CBC at a concentration of between about 0.5 pg/mL and about 16 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 17.5 pg/mL.

9. The combination for use of claim 7, comprises said CBC at a concentration of between about 0.5 pg/mL and about 16 pg/mL and said THC at a concentration of between about 5 pg/mL and about 17.5 pg/mL.

10. The combination for use of claim 7, comprises said CBC and said THC in a concentration according to any one of Table 6A, Table 7A, Table 8A, Table 9A, Table 10A, Table 11 A or combination thereof.

11. The combination for use of claim 7, comprises said CBC at a concentration of between about 0.5 pg/mL and about 12 pg/mL and said THC at a concentration of between about 5 pg/mL and about 15.5 pg/mL.

12. The combination for use of claim 7, comprises at least one of: (i) CBC at a concentration of between about 4 pg/mL and about 8 pg/mL and THC at a concentration of between about 5 pg/mL and about 12.5pg/mL (ii) CBC at a concentration of about 12 pg/mL and THC at a concentration of 10 pg/mL (iii) CBC at a concentration of between about 0.5 pg/mL and about 8 pg/mL and THC at a concentration of about 12.5pg/mL; (iv) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of about 5pg/mL or (v) any combinations thereof.

13. The combination for use of claim 7, comprises said CBC at a concentration of between about 0.5 pg/mL and about 20 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

14. The combination for use of claim 7, comprises said CBC and said THC in a concentration ratio according to any one of 6D, Table 7D, Table 8D, Table 9D, Table 10D, Table 11D or combination thereof.

15. The combination for use of claim 7, comprises at least one of: (i) CBC at a concentration of between about 5 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 25 pg/mL (ii) CBC at a concentration of about 0.5 pg/mL and THC at a concentration of between about 15 pg/mL and 30 pg/mL, (iii) CBC at a concentration of between about 15 pg/mL and about 20 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 5 pg/mL, (iv) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 0.5 pg/mL or (v) any combinations thereof.

16. The combination for use of claim 7, comprises at least one of: (i) CBC at a concentration of 0.5 pg/mL and THC at a concentration of between about 15 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 10 pg/mL and 15 pg/mL, (iii) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 10 pg/mL or (iv) any combinations thereof.

17. The combination for use of any one of claims 1-6, wherein said composition has a synergistic activity on lung cancer.

18. The combination for use of claim 17, comprises said CBC at a concentration of between about 1 pg/mL and about 25 pg/mL and said THC at a concentration of between about 1 pg/mL and about 25 pg/mL.

19. The combination for use of claim 17, comprises said CBC and said THC in a concentration according to any one of Table 12A.

20. The combination for use of claim 17, comprises said CBC and said THC in a concentration ratio according to any one of Table 12D.

21. The combination for use of claim 17, comprises said CBC at a concentration of between about 5 pg/mL and about 25 pg/mL and said THC at a concentration of between about 1 pg/mL and about 20 pg/mL.

22. The combination for use of claim 17, comprises at least one of: (i) CBC at a concentration of between about 10 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 15 pg/mL and 20 pg/mL, (iii) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 5 pg/mL, (iv) CBC at a concentration of about 20 pg/mL and THC at a concentration of between about 1 pg/mL and about 5 pg/mL or (v) any combinations thereof.

23. The combination for use of claim 17, comprises at least one of: (i) CBC at a concentration of about 5 pg/mL and THC at a concentration of about 15 pg/mL, (ii) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 10 pg/mL or (iii) any combinations thereof.

24. The combination for use of any one of claims 1 to 6, wherein said composition has a synergistic activity on colon cancer.

25. The combination for use of claim 24, comprises said CBC at a concentration of between about 1 pg/mL and about 20 pg/mL and said THC at a concentration of between about 1.1 pg/mL and about 25 pg/mL.

26. The combination for use of claim 24, comprises said CBC at a concentration of between about 1 pg/mL and about 15 pg/mL and said THC at a concentration of between about 5 pg/mL and about 20 pg/mL.

27. The combination for use of claim 24, comprises said CBC and said THC in a concentration according to any one of Table 13 A.

28. The combination for use of claim 24, comprises said CBC and said THC in a concentration ratio according to any one of Table 13D.

29. The combination for use of claim 24 comprises at least one of: (i) CBC at a concentration of between about 5 pg/mL and about 10 pg/mL and THC at a concentration of between about 5 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 15 pg/mL and THC at a concentration of between about 5 pg/mL and 15 pg/mL, (iii) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 10 pg/mL, (iv) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 20 pg/mL or (v) any combinations thereof.

30. The combination for use of claim 24, comprises at least one of: (i) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 5 pg/mL and about 10 pg/mL, (ii) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 10 pg/mL or (iii) any combinations thereof.

31. The combination for use of any one of claims 1- 30, being a synthetic composition.

32. The combination for use of any one of claims 1-30, obtained from a cannabis plant extract.

33. The combination for use of any one of claims 1-32, wherein said composition consisting of said CBC and said THC as cannabinoids.

34. A pharmaceutical composition comprising CBC and THC for use in beating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said combination comprises said CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

35. The pharmaceutical composition of claim 34, comprising said CBC and said THC in a concenhation and a concentration ratio according to at least one of Table 1 ± 10 %, Table 2A ± 10 %, Table 2B ± 10 %, Table 3 + 10 %, Table 4 ± 10 % or Table 5 + 10 %.

36. The pharmaceutical composition of claim 34 or 35, having a synergistic effect on at least one of (i) ovarian cancer, (ii) lung cancer, (iii) colon cancer or (iv) combination thereof.

37. The pharmaceutical composition of any one of claims 34-36, comprises at least one of:

(a) one or more of (i) CBC at a concentration of between about 4 pg/mL and about 8 pg/mL and THC at a concentration of between about 5 pg/mL and about 12.5pg/mL (ii) CBC at a concentration of about 12 pg/mL and THC at a concentration of 10 pg/mL (iii) CBC at a concentration of between about 0.5 pg/mL and about 8 pg/mL and THC at a concentration of about 12.5 pg/mL; (iv) CBC at a concentration of between about 0.5 pg/mL and THC at a concentration of about 5pg/mL or (v) any combinations thereof.

(b) one or more of (i) CBC at a concentration of between about 5 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 25 pg/mL (ii) CBC at a concentration of about 0.5 pg/mL and THC at a concentration of between about 15 pg/mL and 30 pg/mL, (iii) CBC at a concentration of between about 15 pg/mL and about 20 pg/mL and THC at a concentration of between about 0.5 pg/mL and about 5 pg/mL, (iv) CBC at a concentration of about 10 pg/mL and THC at a concentration of about 0.5 pg/mL or (v) any combinations thereof.

(c) one or more of (i) CBC at a concentration of between about 5 pg/mL and about 10 pg/mL and THC at a concentration of between about 5 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 15 pg/mL and THC at a concentration of between about 5 pg/mL and 15 pg/mL, (iii) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 10 pg/mL, (iv) CBC at a concentration of about 1 pg/mL and THC at a concentration of about 20 pg/mL or (v) any combinations thereof.

(d) one or more of (i) CBC at a concentration of between about 10 pg/mL and about 20 pg/mL and THC at a concentration of between about 10 pg/mL and about 20 pg/mL (ii) CBC at a concentration of about 5 pg/mL and THC at a concentration of between about 15 pg/mL and 20 pg/mL, (iii) CBC at a concentration of about 15 pg/mL and THC at a concentration of about 5 pg/mL, (iv) CBC at a concentration of about 20 pg/mL and THC at a concentration of between about 1 pg/mL and about 5 pg/mL or (v) any combinations thereof.

38. A method of reducing cell viability, the method comprising contacting the cell with an effective amount of a composition comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

39. The method of claim 38, being an in vivo method and/or an in vitro method.

40. A method of treating, preventing, or delaying the onset of cancer in a subject in need thereof, the method comprising administering to the subject a combination comprising CBC and THC, wherein said CBC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and said THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.

41. The method of any one of claims 38-40, wherein said combination comprises said CBC and said THC in a concentration and a concentration ratio according to at least one of Table 1 ± 10 %, Table 2A ± 10 %, Table 2B ± 10 %, Table 3 + 10 %, Table 4 ± 10 % or Table 5 ± 10 %.

42. The method of any one of claims 38-41, wherein said cell is from a subject suffering from a cancer selected from the group consisting of ovarian cancer, lung cancer, colon cancer.

43. The method of any one of claims 38-41, wherein said combination has a synergistic activity on at least one of (i) ovarian cancer, (ii) lung cancer, (iii) colon cancer or (iv) combination thereof.

44. A kit comprising CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL and optionally comprising instructions for using said CBC and said THC in a combination for treating, preventing, or delaying the onset of cancer in a subject in need thereof.

45. The kit of claim 44, wherein said CBC and said THC is in separate reservoirs.

46. Use of CBC and THC in the preparation of a pharmaceutical composition for treating, preventing, or delaying the onset of cancer in a subject in need thereof, wherein said pharmaceutical composition comprises CBC at a concentration of between about 0.5 pg/mL and about 30 pg/mL and THC is at a concentration of between about 0.5 pg/mL and about 30 pg/mL.