U.S. patent application number 16/434616 was filed with the patent office on 2020-05-07 for anti-tumoural effects of cannabinoid combinations.
This patent application is currently assigned to GW Pharma Limited. The applicant listed for this patent is GW Pharma Limited. Invention is credited to Manuel GUZMAN PASTOR, Mar LORENTE, Fatima RODRIGUEZ, Sofia TORRES, Guillermo VELASCO DIEZ.
Application Number | 20200138771 16/434616 |
Document ID | / |
Family ID | 39638149 |
Filed Date | 2020-05-07 |
United States Patent
Application |
20200138771 |
Kind Code |
A1 |
VELASCO DIEZ; Guillermo ; et
al. |
May 7, 2020 |
ANTI-TUMOURAL EFFECTS OF CANNABINOID COMBINATIONS
Abstract
The invention relates to the use of a combination of
cannabinoids, particularly tetrahydrocannabinol (THC) and
cannabidiol (CBD), in the manufacture of a medicament for use in
the treatment of cancer. In particular the cancer to be treated is
a brain tumour, more particularly a glioma, more particularly still
a glioblastoma multifonne (GBM).
Inventors: |
VELASCO DIEZ; Guillermo;
(Madrid, ES) ; GUZMAN PASTOR; Manuel; (Madrid,
ES) ; LORENTE; Mar; (Madrid, ES) ; TORRES;
Sofia; (Madrid, ES) ; RODRIGUEZ; Fatima;
(Madrid, ES) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GW Pharma Limited |
Cambridge |
|
GB |
|
|
Assignee: |
GW Pharma Limited
Cambridge
GB
|
Family ID: |
39638149 |
Appl. No.: |
16/434616 |
Filed: |
June 7, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16163340 |
Oct 17, 2018 |
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16434616 |
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14079994 |
Nov 14, 2013 |
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16163340 |
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12996124 |
Jan 28, 2011 |
8632825 |
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PCT/GB09/50621 |
Jun 4, 2009 |
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14079994 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/352 20130101; A61K 31/353 20130101; A61K 45/06 20130101;
A61K 31/05 20130101; A61P 35/02 20180101; A61K 31/05 20130101; A61K
31/353 20130101; A61P 43/00 20180101; A61K 2300/00 20130101; A61K
2300/00 20130101 |
International
Class: |
A61K 31/352 20060101
A61K031/352; A61K 31/05 20060101 A61K031/05; A61K 45/06 20060101
A61K045/06; A61K 31/353 20060101 A61K031/353 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 4, 2008 |
GB |
0810195.8 |
Claims
1. Use of a combination of cannabinoids in the manufacture of a
medicament for use in the treatment of cancer.
2. Use as claimed in claim 1, which comprises at least
tetrahydrocannabinol (THC) and cannabidiol (CBD).
3. Use as claimed in claim 2, wherein the THC and CBD are in a
ratio of from between 20:1 to 1:20 (THC:CBD).
4. Use as claimed in claim 2, wherein the THC and CBD are in a
ratio of approximately 1:1.
5. Use as claimed in claim 1, wherein the cannabinoid content is in
the range of between 5 and 100 mg of the total cannabinoids
present.
6. Use as claimed in any claim 1, where each cannabinoid is used at
a level which would be considered sub-optimal if being used
alone.
7. Use as claimed in claim 1, wherein the cancer to be treated is a
brain tumour.
8. Use as claimed in claim 7, wherein the brain tumour is a glioma
tumour.
9. Use as claimed in claim 8, wherein the brain tumour is a
glioblastoma multiforme (GBM).
10. Use as claimed in claim 1, wherein the one or more cannabinoids
are present as plant extracts, as pure compounds, or a combination
of the two.
11. Use as claimed in claim 10, wherein the plant extract is in the
form of a botanical drug substance.
12. Use as claimed in claim 1, wherein the one or more cannabinoids
are administered separately, sequentially, or simultaneously to one
another.
13. A method for preparing a botanical drug substance, wherein the
botanical drug substance comprises THC and CBD, the method
comprising: extracting THC and CBD from a cannabis plant to obtain
a primary extract; and purifying the primary extract by
supercritical or subcritical extraction, vaporization and
chromatography to obtain the botanical drug substance.
Description
[0001] The present invention relates to the use of a combination of
cannabinoids in the manufacture of a medicament for use in the
treatment of cancer. In particular the cancer to be treated is a
brain tumour, more particularly a glioma, more particularly still a
glioblastoma multiforme (GBM) and the preferred cannabinoid
combination comprises tetrahydrocannabinol (THC) and cannabidiol
(CBD).
BACKGROUND TO THE INVENTION
[0002] Cancer a disease in which a group of cells display the
traits of uncontrolled growth. This means that the cells grow and
divide beyond the levels of normal limits. The cells are also able
to invade and destroy surrounding tissues. In addition cancer cells
sometimes also metastasize, meaning that they spread to other
locations in the body via the blood or lymph.
[0003] Most cancers are caused by abnormalities in the genetic
material of the cells. These abnormalities may be due to the
effects of carcinogens. Other cancer-promoting genetic
abnormalities may be randomly acquired through errors in DNA
replication, or are inherited, and thus present in all cells from
birth.
[0004] Genetic abnormalities found in cancer typically affect two
general classes of genes. Cancer-promoting oncogenes are often
activated in cancer cells, giving those cells new properties, such
as hyperactive growth and division, protection against programmed
cell death, loss of respect for normal tissue boundaries, and the
ability to become established in diverse tissue environments.
[0005] Tumour suppressor genes are often inactivated in cancer
cells, resulting in the loss of normal functions in those cells,
such as accurate DNA replication, control over the cell cycle,
orientation and adhesion within tissues, and interaction with
protective cells of the immune system.
[0006] There are many different types of cancer and the cancer is
usually classified according to the type of tissue from which it
originated.
[0007] Cancer is usually treated by one or more of the following:
surgery, chemotherapy, radiation therapy, immunotherapy and
monoclonal antibody therapy. The type of therapy depends upon the
location and grade of the tumour and the stage of the disease.
[0008] Complete removal of the cancer without damage to the rest of
the body is the goal of treatment. Sometimes this can be
accomplished by surgery, but the propensity of cancers to invade
adjacent tissue or to spread to distant sites by microscopic
metastasis often limits its effectiveness. The effectiveness of
chemotherapy is often limited by toxicity to other tissues in the
body. Radiation can also cause damage to normal tissue.
[0009] Cancers are known to affect many areas of the body with the
most common types of cancers including: cancer of the bile duct,
cancer of the bladder, cancer of the bone, cancer of the bowel
(including cancer of the colon and cancer of the rectum), cancer of
the brain, cancer of the breast, cancer of the neuroendocrine
system (commonly known as a carcinoid), cancer of the cervix,
cancer of the eye, cancer of the esophagus, cancer of the head and
neck (this group includes carcinomas that start in the cells that
form the lining of the mouth, nose, throat, ear or the surface
layer covering the tongue), Kaposi's sarcoma, cancer of the kidney,
cancer of the larynx, leukaemia, cancer of the liver, cancer of the
lung, cancer of the lymph nodes, Hodgkin's lymphoma, non-Hodgkin's
lymphoma, melanoma, mesothelioma, myeloma, cancer of the ovary,
cancer of the pancreas, cancer of the penis, cancer of the
prostate, skin cancer, soft tissue sarcomas, cancer of the spinal
cord, cancer of the stomach, testicular cancer, cancer of the
thyroid, cancer of the vagina, cancer of the vulva and cancer of
the uterus.
[0010] A tumour that develops in the brain can destroy or damage
brain cells by producing inflammation, compressing other parts of
the brain, inducing cerebral oedema (brain swelling) and can cause
increases in intracranial pressure (pressure within the skull).
[0011] Each year, approximately 4300 people in the UK are diagnosed
with a brain tumour. A primary brain tumour is a mass created by
the growth or uncontrolled proliferation of cells in the brain.
Malignant primary brain tumours are most likely to cause problems
by spreading into the normal brain tissue which surrounds them and
causing pressure and damage to the surrounding areas of the brain.
These tumours rarely spread outside the brain to other parts of the
body. However, secondary brain tumours occur when cancer cells from
other parts of the body, such as the lung or breast spread to the
brain.
[0012] Surgery is the treatment option of choice for many brain
tumours. Some may be completely excised, but those that are deep or
that infiltrate brain tissue may be debulked rather than
removed.
[0013] Radiation therapy and chemotherapy may be recommended
depending on the type of tumour involved.
[0014] Glioma cell tumours can often be lethal. The characteristic
diffuse infiltrative tumour growth of gliomas often makes the
surgical removal of them impossible and this profoundly complicates
the clinical management of these patients.
[0015] Glioblastoma multiforme (GBM) is the most common and most
aggressive type of primary brain tumour and accounts for 52% of all
primary brain tumour cases and 20% of all intracranial tumours.
[0016] Different approaches are being researched in order to
improve the mortality rate of patients diagnosed with a glioma.
These include therapies that target the glioma cells but leave
normal cells unharmed, methods that limit the spread of the cancer
cells and treatments that block the tumours life-sustaining
molecules.
[0017] One such area of research involves the use of cannabinoids
as anti-tumoural agents.
[0018] Cannabinoids are the active constituents of cannabis plants
and they have been found to demonstrate numerous pharmacological
properties.
[0019] For example EP1177790 (Guzman et al.) describes the
treatment of cerebral tumours by the administration of a natural or
synthetic cannabinoid, specifically THC. It is claimed that
activation of specific receptors leads to selective death of the
transformed cells.
[0020] Recently the cannabinoid CBD has been shown to possess
anti-tumoural properties (Massi et al. 2004). The work described by
this paper describes anti-proliferative effects both in-vitro using
U87 and U373 human glioma cell lines and in-vivo using U87 human
glioma cells subcutaneously implanted to nude mice.
[0021] Malignant gliomas are highly infiltrative and proliferative
tumours, which follow a characteristic pattern of growth. Glioma
cells invade the adjacent normal brain structures and surrounding
large blood vessels.
[0022] In addition the applicant's earlier patent EP1802274
describes the use of the cannabinoid CBD to impede the progress of
cancer cells migrating from their primary tumour location to a
secondary site.
[0023] Furthermore, Medical hypothesis (2006) vol 66, pages 234-246
discusses the physiological and clinical effects of THC and CBD and
presents a rationale for their combination. Under "neoplastic
disease" (page 242) it is acknowledged that THC has cytotoxic
benefits and that CBD has also proven cytostatic/cytotoxic. It is
suggested, given the analgesic effects of the CBD:THC combination
in cancer treatment, the side benefit of THC and CBD in
chemotherapy induced nausea, and these primary effects on tumor
growth and spread that there is a strong rational for additional
clinical trials. However, the generality of this teaching could not
have predicted the benefits that could be achieve in combination in
what would otherwise have been considered sub-optimal (or
ineffective amounts) for the compounds alone.
SUMMARY OF INVENTION
[0024] According to the present invention there is provided the use
of a combination of cannabinoids in the manufacture of a medicament
for use in the treatment of cancer.
[0025] Preferably the cannabinoids comprise at least
tetrahydrocannabinol (THC) and cannabidiol (CBD).
[0026] Preferably the THC and CBD are in a ratio of from between
20:1 to 1:20 (THC:CBD).
[0027] More preferably the THC and CBD are in a ratio of from
between 5:1 to 1:5 (THC:CBD).
[0028] More preferably still, the THC and CBD are in a ratio of
between 2:1 to 1:2, more preferably still, approximately 1:1.
[0029] Each cannabinoid is provided in a therapeutically effect
amount. Dose ranges for the THC and CBD may be determined by
reference to the cannabinoid content which is preferably in the
range of between 5 and 100 mg of the total cannabinoids.
[0030] The cancer to be treated may be a brain tumour.
[0031] Brain tumours are usually classified according to the
location of the tumour and the type of cell that the cancer has
developed from.
[0032] For example different types of brain tumour include:
acoustic neuroma, astrocytoma, CNS lymphoma, ependymoma,
haemangioblastoma, medulloblastoma, meningioma, glioma, mixed
glioma, oligodendroglioma, pineal region tumours and pituitary
tumours.
[0033] Gliomas are tumours of the glial cells; these cells support
and protect nerve cells in the brain. Gliomas comprise nearly half
of all primary brain tumours and a fifth of all primary spinal cord
tumours.
[0034] The cannabinoid combination of the invention is particularly
useful where the brain tumour is a glioma tumour, more particularly
glioblastoma multiforme (GBM).
[0035] The one or more cannabinoids may be present as plant
extracts, as pure compounds, or a combination of the two.
[0036] A plant extract is defined as an extract from a plant
material as described by the Guidance for Industry Botanical Drug
Products Draft Guidance, August 2000, US Department of Health and
Human Services, Food and Drug Administration Centre for Drug
Evaluation and Research.
[0037] Plant material is defined as a plant or plant part (e.g.
bark, wood, leaves, stems, roots, flowers, fruits, seeds, berries
or parts thereof) as well as exudates.
[0038] More preferably the plant extract is in the form of a
botanical drug substance.
[0039] Botanical drug substances which are derived from cannabis
plants include primary extracts prepared by such processes as for
example, maceration, percolation, extraction with solvents such as
C1 to C5 alcohols (e.g. ethanol), Norflurane (HFA134a), HFA227,
liquid carbon dioxide under pressure and extraction using a hot
gas. A primary extract may be further purified by supercritical or
subcritical extraction, vaporisation and chromatography. When
solvents such as those listed above are used the resultant extract
may contain non-specific lipid-soluble material. This can be
removed by a variety of processes including winterisation, which
involves chilling to -20.degree. C. followed by filtration to
remove waxy ballast, extraction with liquid carbon dioxide and by
distillation.
[0040] Botanical drug substances are formulated into Botanical Drug
Products which are defined in the Guidance for Industry Botanical
Drug Products Draft Guidance, August 2000, US Department of Health
and Human Services, Food and Drug Administration Centre for Drug
Evaluation and Research as: "A botanical product that is intended
for use as a drug; a drug product that is prepared from a botanical
drug substance."
[0041] The one or more cannabinoids may be administered separately,
sequentially or simultaneously to one another.
[0042] Certain aspects of this invention are further described, by
way of example only, with reference to the accompanying drawings in
which:
[0043] FIG. 1 is a bar chart showing the cell viability of human
U87 MG astrocytoma cells after treatment with THC, CBD or a
combination of THC and CBD in comparison to a control;
[0044] FIGS. 2a and 2b are bar charts showing in vivo cell
viability data at different concentrations on two cell lines, U87MG
(FIG. 2a) and T98G (FIG. 2b); and
[0045] FIGS. 3a, 3b and 3c provide data suggestive of the mechanism
of action of the combination for U87MG cells.
SPECIFIC DESCRIPTION
[0046] The following examples describe experiments undertaken to
ascertain the effect of combinations of cannabinoids as
anti-tumoural agents.
Example 1: The Effect of THC and CBD at Inhibiting Cancer Cell
Growth In Vitro
[0047] Tetrahydrocannabinol (THC) and cannabidiol (CBD) in the form
of cannabis plant extracts were dissolved in ethanol to a
concentration of 100 mM this was stored at -20.degree. C. until
required.
[0048] Before use the cannabis plant extracts were further diluted
to the desired concentration, ensuring that the concentration of
ethanol was below 0.001%.
[0049] U87 human glioma cells were used throughout this experiment.
The cells were maintained at 37.degree. C. in a humidified
atmosphere with 5% CO.sub.2 and 95% air.
[0050] Cells were cultured in a 75 cm.sup.2 culture flask in
Dulbecco's Modified Eagle Medium (DMEM), which had been
supplemented with 4 mM L-glutamine, 100 units/ml penicillin, 100
mg/ml streptomycin, 1% sodium pyruvate, 1% non-essential amino
acids and 10% heat-inactivated fetal bovine serum.
[0051] The viability of the human U87 MG astrocytoma cells were
examined at various cannabinoid concentrations. The THC and CBD
extracts were compared against pure THC and CBD.
Results:
TABLE-US-00001 [0052] TABLE 1 Cell viability of human U87 MG
astrocytoma cells in culture IC50 .mu.M IC50 .mu.M IC50 .mu.M
(equivalent of (pure (cannabis pure in cannabis cannabinoids) plant
extract) plant extract) THC 0.37 0.64 0.43 CBD 0.47 0.72 0.47
[0053] As can be seen from Table 1 above the THC and CBD extracts
compare very favourably in activity to their corresponding pure
compounds, when the amount of cannabinoid in the extract is
adjusted to an equivalent amount of pure compound.
[0054] This shows that THC and CBD and their extracts are effective
in inhibiting glioma cell growth.
Example 2: The Effect of a Combination of THC and CBD Extracts at
Inhibiting Cancer Cell Growth In Vitro
[0055] This experiment tested whether a combination of THC and CBD
extracts were as effective at inhibiting cell growth as the
extracts alone.
[0056] The methods used were as described in Example 1 above.
Results:
[0057] FIG. 1 details a bar chart describing the cell viability of
human U87 MG astrocytoma cells versus the THC and CBD extracts
alone and in combination with one another.
[0058] As can be seen when the THC and CBD are used in combination
the cell viability is significantly reduced in comparison to the
cell viability after treatment with either THC or CBD alone.
[0059] This data suggests that the cannabinoids THC and CBD would
be more effective in the treatment of tumours when used in
combination.
Example 3: The Effect of a Combination of THC and CBD at Inhibiting
Cancer Cell Growth In Vivo
[0060] This experiment tested whether the combination of THC and
CBD extracts were also effective in vivo.
[0061] Human U87 MG astrocytoma cells were xenografted to nude mice
and the test compounds were injected peritumourally at a
concentration of 15 mg/kg per day.
Results:
TABLE-US-00002 [0062] TABLE 2 Tumour volume relative to zero time
following 15 days of treatment Tumour volume Vehicle 9.2 .+-. 0.6
Pure THC 5.1 .+-. 0.4 THC extract 6.6 .+-. 0.3 THC:CBD (1:1)
extract 4.8 .+-. 0.3
[0063] As can be observed in Table 2 above the tumour volume after
treatment with the 1:1 combination of THC and CBD extracts is
significantly superior to the treatment with either the pure THC or
the THC extract alone.
[0064] This data suggests that the cannabinoids THC and CBD would
be more effective in the treatment of tumours when used in
combination.
Example 4: Effect of Cannabinoid Concentration on Cell Viability in
Two Different Cell Lines
[0065] The action of THC, CBD, and a 1:1 ratio mix of THC and CBD
were studied at different concentrations on two cell lines: U87MG
and T98G. The cell viability data is illustrated in FIGS. 2a and
2b.
[0066] Referring to FIG. 2a it will be seen that
ineffective/sub-optimal doses of THC and CBD at 0.1 ug/ml and 0.25
ug/ml (greater than 90% cell viability) gave way to a statistically
significant decrease in cell viability in combination (SAT), which
data showed a dose dependant relationship with increased
concentration (greater cyto-toxicity at 0.25 ug/ml).
[0067] Similar results were obtained with cell line T98G, (an
alternative human glioma cell line) as is shown in FIG. 2b.
Example 5: Investigation of Mechanism of Action
[0068] THC is known to induce cell death using a signalling route
involving the gene ATG1 and pan-caspase. The results of an
investigation looking at S6 phosphorylation, LC3 lipidation and the
effect of an ATG1 and a pan-caspase inhibitor are shown in FIGS.
3a, 3b and 3c respectively.
[0069] It can be seen from FIG. 3a that the THC:CBD combination
(compare to control C): [0070] Inhibits mTORC1 activity (as
determined by the levels of S6 phosphorylation); and [0071]
Promotes accumulation of the lipidated form LC3 (a hall mark of
autophagy).
[0072] FIG. 3b shows that silencing the essential autophagy gene
ATG1, with a selective (siATG10) siRNA inhibitor reduces induced
cell death compared to cells transfected with a control siC.
[0073] Finally, FIG. 3c shows that cells treated with the
pan-capase inhibitor Z-VAD also prevent induced cell death.
* * * * *