U.S. patent application number 17/050956 was filed with the patent office on 2021-02-25 for cannabidiol preparations and its uses.
The applicant listed for this patent is GW Research Limited. Invention is credited to James BRODLE, Royston GRAY, Geoffrey GUY, Volker KNAPPERTZ, Katarzyna LACH-FALCONE, Rohini Rajyalaxmi RANA, Alan SUTTON, Benjamin WHALLEY, Marie WOOLLEY-ROBERTS.
Application Number | 20210052512 17/050956 |
Document ID | / |
Family ID | 1000005235464 |
Filed Date | 2021-02-25 |
View All Diagrams
United States Patent
Application |
20210052512 |
Kind Code |
A1 |
GUY; Geoffrey ; et
al. |
February 25, 2021 |
CANNABIDIOL PREPARATIONS AND ITS USES
Abstract
Cannabidiol (CBD) is a cannabinoid designated chemically as
2-[(1R,6R)-3-Methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]-5-pentyl-1,3-b-
enzenediol. Its empirical formula is C.sub.21H.sub.30O.sub.2 and
its molecular weight is 314.46. CBD is a cannabinoid that naturally
occurs in the Cannabis sativa L. plant. CBD is a white to pale
yellow crystalline solid which is insoluble in water and soluble in
organic solvents. The present invention encompasses the surprising
recognition that certain CBD preparations which are prepared from a
botanical origin are more effective in treating diseases or
disorders than preparations of CBD which are synthetic or purified
to the extent no other impurities in the form of other cannabinoids
are present. Prior CBD compositions have been prepared such that no
psychoactive components, e.g., tetrahydrocannabinol (THC), remain
in the final CBD preparation. Surprisingly, the absence of such
minor impurities reduces the efficacy of CBD treatment. Such CBD
preparations are characterized by chemical components and/or
functional properties that distinguish them from prior CBD
compositions. One or more components of the preparations described
herein provide an unexpectedly synergistic effect when utilized in
combination.
Inventors: |
GUY; Geoffrey; (Cambridge,
GB) ; KNAPPERTZ; Volker; (Cambridge, GB) ;
WHALLEY; Benjamin; (Cambridge, GB) ; WOOLLEY-ROBERTS;
Marie; (Cambridge, GB) ; BRODLE; James;
(Cambridge, GB) ; LACH-FALCONE; Katarzyna;
(Cambridge, GB) ; SUTTON; Alan; (Cambridge,
GB) ; GRAY; Royston; (Cambridge, GB) ; RANA;
Rohini Rajyalaxmi; (Cambridge, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GW Research Limited |
Cambridge |
|
GB |
|
|
Family ID: |
1000005235464 |
Appl. No.: |
17/050956 |
Filed: |
April 26, 2019 |
PCT Filed: |
April 26, 2019 |
PCT NO: |
PCT/GB2019/051173 |
371 Date: |
October 27, 2020 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 36/185 20130101;
A61P 25/08 20180101; A61K 31/352 20130101; A61P 25/18 20180101;
A61K 31/05 20130101 |
International
Class: |
A61K 31/05 20060101
A61K031/05; A61K 31/352 20060101 A61K031/352; A61K 36/185 20060101
A61K036/185; A61P 25/08 20060101 A61P025/08; A61P 25/18 20060101
A61P025/18 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 27, 2018 |
GB |
1806953.4 |
Claims
1. A cannabidiol (CBD) preparation characterized in that it
comprises greater than or equal to 98% (w/w) CBD and less than or
equal to 2% (w/w) other cannabinoids, wherein the less than or
equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC.
2. A cannabidiol (CBD) preparation characterized in that it
comprises greater than or equal to 98% (w/w) CBD and less than or
equal to 2% (w/w) other cannabinoids, wherein the less than or
equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use as a medicament.
3. A cannabidiol (CBD) preparation characterized in that it
comprises greater than or equal to 98% (w/w) CBD and less than or
equal to 2% (w/w) other cannabinoids, wherein the less than or
equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
neurodevelopmental diseases and conditions.
4. A cannabidiol (CBD) preparation characterized in that it
comprises greater than or equal to 98% (w/w) CBD and less than or
equal to 2% (w/w) other cannabinoids, wherein the less than or
equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
epilepsy.
5. A cannabidiol (CBD) preparation characterized in that it
comprises greater than or equal to 98% (w/w) CBD and less than or
equal to 2% (w/w) other cannabinoids, wherein the less than or
equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
schizophrenia.
6. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the preparation comprises not more than
1.5% (w/w) THC based on total amount of cannabinoid in the
preparation.
7. A cannabidiol (CBD) preparation as claimed in claim 6, wherein
the preparation comprises about 0.01% to about 0.1% (w/w) THC based
on total amount of cannabinoid in the preparation.
8. A cannabidiol (CBD) preparation as claimed in claim 6, wherein
the preparation comprises about 0.02% to about 0.05% (w/w) THC
based on total amount of cannabinoid in the preparation.
9. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the mixture of trans-THC and cis-THC is
present at a ratio of about 3.6:1 trans-THC:cis-THC.
10. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the mixture of trans-THC and cis-THC is
present at a ratio of about 0.8:1 trans-THC:cis-THC.
11. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the preparation comprises about 0.1% to
about 0.15% (w/w) CBD-C1 based on total amount of cannabinoid in
the preparation.
12. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the preparation comprises about 0.2% to
about 0.8% (w/w) CBDV based on total amount of cannabinoid in the
preparation.
13. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein the preparation comprises about 0.3% to
about 0.4% (w/w) CBD-C4 based on total amount of cannabinoid in the
preparation.
14. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein at least a portion of at least one of the
cannabinoids present in the CBD preparation is isolated from
Cannabis plant material.
15. A cannabidiol (CBD) preparation as claimed in claim 14, wherein
at least a portion of the CBD present in the CBD preparation is
isolated from Cannabis plant material.
16. A cannabidiol (CBD) preparation as claimed in claim 14, wherein
at least a portion of the THC present in the CBD preparation is
isolated from Cannabis plant material.
17. A cannabidiol (CBD) preparation as claimed in claim 14, wherein
at least a portion of the CBD-C1 present in the CBD preparation is
isolated from Cannabis plant material.
18. A cannabidiol (CBD) preparation as claimed in claim 14, wherein
at least a portion of the CBDV present in the CBD preparation is
isolated from Cannabis plant material.
19. A cannabidiol (CBD) preparation as claimed in claim 14, wherein
at least a portion of the CBD-C4 present in the CBD preparation is
isolated from Cannabis plant material.
20. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein substantially all of at least one of the
cannabinoids present in the CBD preparation is isolated from
Cannabis plant material.
21. A cannabidiol (CBD) preparation as claimed in claim 20, wherein
substantially all of the CBD present in the CBD preparation is
isolated from Cannabis plant material.
22. A cannabidiol (CBD) preparation as claimed in claim 20, wherein
substantially all of the THC present in the CBD preparation is
isolated from Cannabis plant material.
23. A cannabidiol (CBD) preparation as claimed in claim 20, wherein
substantially all of the CBD-C1 present in the CBD preparation is
isolated from Cannabis plant material.
24. A cannabidiol (CBD) preparation as claimed in claim 20, wherein
substantially all of the CBDV present in the CBD preparation is
isolated from Cannabis plant material.
25. A cannabidiol (CBD) preparation as claimed in claim 20, wherein
substantially all of the CBD-C4 present in the CBD preparation is
isolated from Cannabis plant material.
26. A cannabidiol (CBD) preparation as claimed in any of the
preceding claims, wherein substantially all of the cannabinoids
present in the CBD preparation are isolated from Cannabis plant
material.
27. A cannabidiol (CBD) preparation as claimed in any of claims 14
to 26, wherein the Cannabis plant material is from a Cannabis
sativa, Cannabis indica, or Cannabis ruderalis plant.
28. A cannabidiol (CBD) preparation as claimed in any of claims 14
to 26, wherein the Cannabis plant is a high-CBD containing Cannabis
chemotype.
29. A cannabidiol (CBD) preparation as claimed in any of claims 1
to 13, wherein at least a portion of at least one of the
cannabinoids present in the CBD preparation is prepared
synthetically.
30. A cannabidiol (CBD) preparation as claimed in claim 29, wherein
at least a portion of the CBD present in the CBD preparation is
prepared synthetically.
31. A cannabidiol (CBD) preparation as claimed in claim 29, wherein
at least a portion of the THC present in the CBD preparation is
prepared synthetically.
32. A cannabidiol (CBD) preparation as claimed in claim 29, wherein
at least a portion of the CBD-C1 present in the CBD preparation is
prepared synthetically.
33. A cannabidiol (CBD) preparation as claimed in claim 29, wherein
at least a portion of the CBDV present in the CBD preparation is
prepared synthetically.
34. A cannabidiol (CBD) preparation as claimed in claim 29, wherein
at least a portion of the CBD-C4 present in the CBD preparation is
prepared synthetically.
35. A cannabidiol (CBD) preparation as claimed in any of claims 1
to 13, wherein substantially all of at least one of the
cannabinoids present in the CBD preparation is prepared
synthetically.
36. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all the CBD present in the CBD preparation is
prepared synthetically.
37. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all the THC present in the CBD preparation is
prepared synthetically.
38. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all the CBD-C1 present in the CBD preparation is
prepared synthetically.
39. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all the CBDV present in the CBD preparation is
prepared synthetically.
40. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all the CBD-C4 present in the CBD preparation is
prepared synthetically.
41. A cannabidiol (CBD) preparation as claimed claim 35, wherein
substantially all of the cannabinoids present in the CBD
preparation are prepared synthetically.
42. A cannabidiol (CBD) preparation as claimed in claim 3, wherein
the neurodegenerative disease or disorder is Alzheimer's disease;
Parkinson's disease; essential tremor; amyotrophic lateral
sclerosis (ALS); Huntington's disease; Friedreich's ataxia;
multiple sclerosis; frontotemporal dementia; prion disease; Lewy
body dementia; progressive supranuclear palsy; vascular dementia;
normal pressure hydrocephalus; traumatic spinal cord injury; HIV
dementia; alcohol induced neurotoxicity; Down's syndrome; movement
disorders of the central and/or peripheral nervous system; motor
neurone diseases (MND); spinal muscular atrophy; or any other
related neurological or psychiatric neurodegenerative disease;
brain damage; brain injury; brain dysfunction; dysgraphia;
dysarthria; apraxia; agnosia; amnesia; dizziness; vertigo; coma;
stroke; spinal cord damage; spinal cord injury; spinal cord
disorders; central neuropathy; peripheral neuropathy; cranial nerve
disorder; trigeminal neuralgia; tumors of the nervous system;
infections of the brain or spinal cord; encephalitis; meningitis;
prion disease; complex regional pain syndrome; an autonomic nervous
system disorder; autonomic neuropathy; dysautonomia; postural
orthostatic tachycardia syndrome (POTS); neurocardiogenic syncope
(NCS); multiple system atrophy (MSA); hereditary sensory and
autonomic neuropathy (HSAN); Holmes-Adie syndrome (HAS); a sleep
disorder; narcolepsy; pain; migraine; cluster headache; tension
headache; back pain; lower back pain; neck pain; neuropathic pain;
cancer pain; allodynia; arthritic pain; inflammatory pain; a
neuropsychiatric disorder; attention deficit hyperactivity
disorder; autism; Tourette's Syndrome; obsessive compulsive
disorder; an autism spectrum disorder; Rett syndrome; Fragile X
syndrome; Angelman syndrome; hyperkinetic disorder; mitochondrial
disease; dystonia; a cancer; brain cancer; glioma; breast cancer;
liver cancer; lung cancer; pancreatic cancer; melanoma; ovarian
cancer; gastric cancer; renal cancer; bladder cancer; addiction;
nicotine addiction; smoking; alcohol addiction; drug addiction;
Cannabis use disorder; a mental disorder; post-traumatic stress
disorder; anxiety; early psychosis; schizophrenia; a cognitive
disorder; stroke; cardiac ischemia; coronary artery disease;
thromboembolism; myocardial infarction; ischemic related disease; a
gastrointestinal disorder; inflammatory bowel disease; Crohn's
disease; ulcerative colitis; nausea; vomiting; emesis; motion
sickness; chemotherapy induced nausea; chemotherapy induced nausea
vomiting; inflammation; arthritis; rheumatoid arthritis;
osteoarthritis; diabetes; high blood pressure; poor insulin
control; appetite suppression; anorexia; neonatal hypoxic-ischemic
encephalopathy (NHIE); a degenerative skeletal muscle disease; or
Duchenne muscular dystrophy (DMD).
43. A cannabidiol (CBD) preparation as claimed in claim 4, wherein
the epilepsy is Dravet syndrome, Lennox Gastaut syndrome, febrile
infection related epilepsy syndrome (FIRES), Doose syndrome, Sturge
Weber syndrome, CDKL5 mutation; Aicardi syndrome; bilateral
polymicrogyria; Dup15q; SNAP25; benign rolandic epilepsy; juvenile
myoclonic epilepsy; infantile spasm (West syndrome); and
Landau-Kleffner syndrome, refractory epilepsy, juvenile spasms,
West syndrome, infantile spasms, refractory infantile spasms,
tuberous sclerosis complex (TSC); neurogenetic storage disorder,
neuronal ceroid lipofuscinoses (NCL), Batten disease, brain
abnormality, atonic, idiopathic, absence seizure, partial seizure,
simple partial seizure, or complex partial seizure.
Description
FIELD OF THE INVENTION
[0001] Cannabidiol (CBD) is a cannabinoid designated chemically as
2-[(1R,6R)-3-Methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]-5-pentyl-1,3-b-
enzenediol. Its empirical formula is C.sub.21H.sub.30O.sub.2 and
its molecular weight is 314.46. CBD is a cannabinoid that naturally
occurs in the Cannabis sativa L. plant. CBD is a white to pale
yellow crystalline solid which is insoluble in water and soluble in
organic solvents.
[0002] The present invention encompasses the surprising recognition
that certain CBD preparations which are prepared from a botanical
origin are more effective in treating diseases or disorders than
preparations of CBD which are synthetic or purified to the extent
no other impurities in the form of other cannabinoids are
present.
[0003] Prior CBD compositions have been prepared such that no
psychoactive components, e.g., tetrahydrocannabinol (THC), remain
in the final CBD preparation. Surprisingly, the absence of such
minor impurities reduces the efficacy of CBD treatment.
[0004] Such CBD preparations are characterized by chemical
components and/or functional properties that distinguish them from
prior CBD compositions. One or more components of the preparations
described herein provide an unexpectedly synergistic effect when
utilized in combination.
BACKGROUND TO THE INVENTION
[0005] Cannabinoids are natural and synthetic compounds
structurally or pharmacologically related to the constituents of
the Cannabis plant or to the endogenous agonists (endocannabinoids)
of the cannabinoid receptors CB1 or CB2. The only way in nature in
which these compounds are produced is by the Cannabis plant.
Cannabis is a genus of flowering plants in the family Cannabaceae,
comprising the species Cannabis sativa, Cannabis indica, and
Cannabis ruderalis (sometimes considered as part of Cannabis
sativa).
[0006] Cannabis plants comprise a highly complex mixture of
compounds. At least 568 unique molecules have been identified.
Among these compounds are cannabinoids, terpenoids, sugars, fatty
acids, flavonoids, other hydrocarbons, nitrogenous compounds, and
amino acids.
[0007] Cannabinoids exert their physiological effects through a
variety of receptors including, but not limited to, adrenergic
receptors, cannabinoid receptors (CB1 and CB2), GPR55, GPR3, or
GPR5. The principle cannabinoids present in Cannabis plants are
cannabinoid acids tetrahydrocannabinolic acid (THCA) and
cannabidiolic acid (CBDA) with small amounts of their respective
neutral (decarboxylated) cannabinoids. In addition, Cannabis may
contain lower levels of other minor cannabinoids.
[0008] Crude extracts from Cannabis plants containing CBD have been
used by patients suffering from diseases and disorders. However,
such crude products are unsuitable for use in pharmaceutical
formulations. Those seeking to prepare more consistent CBD
preparations for use in treating diseases or disorders have made a
concerted effort to either prepare CBD synthetically or attempt to
remove all compounds other than CBD, particularly psychoactive
compounds such as THC, from plant derived cannabinoids.
[0009] The present invention encompasses the surprising discovery
that particular preparations comprising CBD have an improved
therapeutic efficacy in comparison to synthetic preparations of CBD
which comprise no minor cannabinoid impurities and crude extracts
which have higher levels of the minor cannabinoid impurities.
[0010] As stated, cannabinoids are a class of compounds which may
be derived naturally from the Cannabis plant or produced
synthetically via chemical synthesis.
[0011] More than 100 different cannabinoids produced by Cannabis
have been identified as described in Handbook of Cannabis, Roger
Pertwee, Chapter 1, pages 3 to 15. These cannabinoids can be split
into different groups as follows: phytocannabinoids;
endocannabinoids and synthetic cannabinoids (which may be novel
cannabinoids or synthetically produced versions of
phytocannabinoids or endocannabinoids).
[0012] Phytocannabinoids are cannabinoids that originate from
nature and can be found in the Cannabis plant. Phytocannabinoids
can be isolated from plants to produce a highly purified extract.
Phytocannabinoids may be obtained as either the neutral
(decarboxylated form) or the carboxylic acid form depending on the
method used to extract the cannabinoids from plant material. For
example, it is known that heating the carboxylic acid form will
cause most of the carboxylic acid form to decarboxylate into the
neutral form. Phytocannabinoids can only be produced from plants,
however versions of phytocannabinoids may be produced synthetically
via chemical synthesis.
[0013] Endocannabinoids are endogenous lipid-based retrograde
neurotransmitters that bind to cannabinoid receptors, and
cannabinoid receptor proteins that are expressed throughout the
mammalian central nervous system (including the brain) and
peripheral nervous system. The endocannabinoid system is involved
in regulating a variety of physiological and cognitive processes
including fertility, pregnancy, during pre- and postnatal
development, appetite, pain-sensation, mood, and memory, and in
mediating the pharmacological effects of Cannabis.
[0014] Synthetic cannabinoids are compounds that have a
cannabinoid-like structure and are manufactured using chemical
means rather than by the plant.
[0015] Certain cannabinoids are described in more detail below.
[0016] Cannabidiol (CBD) is a major cannabinoid constituent of
Cannabis species, such as the hemp plant (Cannabis sativa). Unlike
other cannabinoids, such as THC, cannabidiol does not bind to CB1
or CB2, or its binding to the receptors is negligible in terms of
inducing a pharmacological effect. Thus, cannabidiol does not cause
the central or peripheral nervous system effects mediated by the
CB1 or CB2 receptors. CBD has little or no psychotropic
(cannabimimetic) activity and its molecular structure and
properties are substantially different from those of other
cannabinoids.
[0017] Cannabidiol administration has been the subject of research
in an attempt to provide an alternative treatment for various
diseases and disorders which may respond to such treatment.
[0018] Tetrahydrocannabinol (THC) is the principal psychoactive
constituent of Cannabis. THC is a partial agonist at the CB1 and
CB2 receptors. Synthetic THC or dronabinol is approved for the
treatment of loss of appetite in AIDS patients and nausea and
vomiting caused by cancer chemotherapy. The cannabimimetic side
effects caused by THC include feeling high, nausea, vomiting,
anxiety, depression and weakness.
[0019] Of the over 100 natural cannabinoids identified in Cannabis
sativa, seven have been classified as CBD-type compounds, these
cannabinoids have the same absolute configuration as CBD. These
are: CBD, Cannabidiolic acid (CBDA), Cannabidivarin (CBDV),
Cannabidivarin acid (CBDVA), Cannabidiol-C1 (CBD-C1),
Cannabidiol-C4 (CBD-C4) and Cannabidiol monomethyl ether
(CBDM).
[0020] Cannabidiolic acid (CBDA) is the main form in which CBD
exists in the Cannabis plant. It is converted into CBD after
decarboxylation.
[0021] Cannabidiol-C1 (CBD-C1) also known as cannabidiorcol is a
homolog of CBD, with the side-chain shortened by four methylene
bridges. CBD-C1 occurs naturally in plants producing CBD in minor
quantities.
[0022] Cannabidivarin (CBDV) is a homolog of CBD, with the
side-chain shortened by two methylene bridges. CBDV is a
non-psychoactive cannabinoid and has been shown to have
anti-convulsant activity in a mouse model of epilepsy
[0023] Cannabidiol-C4 (CBD-C4) also known as nor-cannabidiol is a
homolog of CBD, with the side-chain shortened by one methylene
bridge. CBD-C4 occurs naturally in plants producing CBD in minor
quantities.
[0024] The present invention demonstrates an increased efficacy of
a botanically derived purified CBD preparation which comprises
minor amounts of the cannabinoids CBD-C1, CBDV, CBD-C4 and THC over
a synthetic CBD which does not comprise minor amounts of
cannabinoids. These data are particularly surprising particularly
given the fact that the concentration of CBD within the botanically
derived purified CBD preparation and the synthetic preparation were
the same. The invention further discloses the difference in the
physicochemical properties of a botanically derived purified CBD
and a synthetic CBD.
BRIEF SUMMARY OF THE DISCLOSURE
[0025] In accordance with a first aspect of the present invention
there is provided a cannabidiol (CBD) preparation characterized in
that it comprises greater than or equal to 98% (w/w) CBD and less
than or equal to 2% (w/w) other cannabinoids, wherein the less than
or equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC.
[0026] In accordance with a second aspect of the present invention
there is provided a cannabidiol (CBD) preparation characterized in
that it comprises greater than or equal to 98% (w/w) CBD and less
than or equal to 2% (w/w) other cannabinoids, wherein the less than
or equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use as a medicament.
[0027] In accordance with a third aspect of the present invention
there is provided a cannabidiol (CBD) preparation characterized in
that it comprises greater than or equal to 98% (w/w) CBD and less
than or equal to 2% (w/w) other cannabinoids, wherein the less than
or equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
neurodevelopmental diseases and conditions.
[0028] In accordance with a fourth aspect of the present invention
there is provided a cannabidiol (CBD) preparation characterized in
that it comprises greater than or equal to 98% (w/w) CBD and less
than or equal to 2% (w/w) other cannabinoids, wherein the less than
or equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
epilepsy.
[0029] In accordance with a fifth aspect of the present invention
there is provided a cannabidiol (CBD) preparation characterized in
that it comprises greater than or equal to 98% (w/w) CBD and less
than or equal to 2% (w/w) other cannabinoids, wherein the less than
or equal to 2% (w/w) other cannabinoids comprise the cannabinoids
tetrahydrocannabinol (THC); cannabidiol-C1 (CBD-C1); cannabidivarin
(CBDV); and cannabidiol-C4 (CBD-C4), and wherein the THC is present
as a mixture of trans-THC and cis-THC for use in the treatment of
schizophrenia.
[0030] Preferably the preparation comprises not more than 1.5%
(w/w) THC based on total amount of cannabinoid in the preparation.
More preferably the preparation comprises about 0.01% to about 0.1%
(w/w) THC based on total amount of cannabinoid in the preparation.
More preferably still the preparation comprises about 0.02% to
about 0.05% (w/w) THC based on total amount of cannabinoid in the
preparation.
[0031] In an embodiment of the invention the mixture of trans-THC
and cis-THC is present at a ratio of about 3.6:1 trans-THC:cis-THC.
In a further embodiment of the invention the mixture of trans-THC
and cis-THC is present at a ratio of about 0.8:1
trans-THC:cis-THC.
[0032] Preferably the preparation comprises about 0.1% to about
0.15% (w/w) CBD-C1 based on total amount of cannabinoid in the
preparation.
[0033] Preferably the preparation comprises about 0.2% to about
0.8% (w/w) CBDV based on total amount of cannabinoid in the
preparation.
[0034] Preferably the preparation comprises about 0.3% to about
0.4% (w/w) CBD-C4 based on total amount of cannabinoid in the
preparation.
[0035] In one embodiment at least a portion of at least one of the
cannabinoids present in the CBD preparation is isolated from
Cannabis plant material.
[0036] Preferably at least a portion of the CBD present in the CBD
preparation is isolated from Cannabis plant material.
[0037] Preferably at least a portion of the THC present in the CBD
preparation is isolated from Cannabis plant material.
[0038] Preferably at least a portion of the CBD-C1 present in the
CBD preparation is isolated from Cannabis plant material.
[0039] Preferably at least a portion of the CBDV present in the CBD
preparation is isolated from Cannabis plant material.
[0040] Preferably at least a portion of the CBD-C4 present in the
CBD preparation is isolated from Cannabis plant material.
[0041] In a further embodiment of the invention substantially all
of at least one of the cannabinoids present in the CBD preparation
is isolated from Cannabis plant material.
[0042] Preferably substantially all of the CBD present in the CBD
preparation is isolated from Cannabis plant material.
[0043] Preferably substantially all of the THC present in the CBD
preparation is isolated from Cannabis plant material.
[0044] Preferably substantially all of the CBD-C1 present in the
CBD preparation is isolated from Cannabis plant material.
[0045] Preferably all of the CBDV present in the CBD preparation is
isolated from cannabis plant material.
[0046] Preferably all of the CBD-C4 present in the CBD preparation
is isolated from Cannabis plant material.
[0047] In a further embodiment of the invention substantially all
of the cannabinoids present in the CBD preparation are isolated
from Cannabis plant material.
[0048] Preferably the Cannabis plant material is from a Cannabis
sativa, Cannabis indica, or Cannabis ruderalis plant.
[0049] Preferably the Cannabis plant is a high-CBD containing
cannabis chemotype.
[0050] In a further embodiment of the invention at least a portion
of at least one of the cannabinoids present in the CBD preparation
is prepared synthetically.
[0051] Preferably at least a portion of the CBD present in the CBD
preparation is prepared synthetically.
[0052] Preferably at least a portion of the THC present in the CBD
preparation is prepared synthetically.
[0053] Preferably at least a portion of the CBD-C1 present in the
CBD preparation is prepared synthetically.
[0054] Preferably at least a portion of the CBDV present in the CBD
preparation is prepared synthetically.
[0055] Preferably at least a portion of the CBD-C4 present in the
CBD preparation is prepared synthetically.
[0056] In a further embodiment of the invention substantially all
of at least one of the cannabinoids present in the CBD preparation
is prepared synthetically.
[0057] Preferably substantially all the CBD present in the CBD
preparation is prepared synthetically.
[0058] Preferably substantially all the THC present in the CBD
preparation is prepared synthetically.
[0059] Preferably substantially all the CBD-C1 present in the CBD
preparation is prepared synthetically.
[0060] Preferably substantially all the CBDV present in the CBD
preparation is prepared synthetically.
[0061] Preferably substantially all the CBD-C4 present in the CBD
preparation is prepared synthetically.
[0062] Preferably substantially all of the cannabinoids present in
the CBD preparation are prepared synthetically.
[0063] In a further embodiment of the invention the
neurodegenerative disease or disorder is Alzheimer's disease;
Parkinson's disease; essential tremor; amyotrophic lateral
sclerosis (ALS); Huntington's disease; Friedreich's ataxia;
multiple sclerosis; frontotemporal dementia; prion disease; Lewy
body dementia; progressive supranuclear palsy; vascular dementia;
normal pressure hydrocephalus; traumatic spinal cord injury; HIV
dementia; alcohol induced neurotoxicity; Down's syndrome; movement
disorders of the central and/or peripheral nervous system; motor
neurone diseases (MND); spinal muscular atrophy; or any other
related neurological or psychiatric neurodegenerative disease;
brain damage; brain injury; brain dysfunction; dysgraphia;
dysarthria; apraxia; agnosia; amnesia; dizziness; vertigo; coma;
stroke; spinal cord damage; spinal cord injury; spinal cord
disorders; central neuropathy; peripheral neuropathy; cranial nerve
disorder; trigeminal neuralgia; tumors of the nervous system;
infections of the brain or spinal cord; encephalitis; meningitis;
prion disease; complex regional pain syndrome; an autonomic nervous
system disorder; autonomic neuropathy; dysautonomia; postural
orthostatic tachycardia syndrome (POTS); neurocardiogenic syncope
(NCS); multiple system atrophy (MSA); hereditary sensory and
autonomic neuropathy (HSAN); Holmes-Adie syndrome (HAS); a sleep
disorder; narcolepsy; pain; migraine; cluster headache; tension
headache; back pain; lower back pain; neck pain; neuropathic pain;
cancer pain; allodynia; arthritic pain; inflammatory pain; a
neuropsychiatric disorder; attention deficit hyperactivity
disorder; autism; Tourette's Syndrome; obsessive compulsive
disorder; an autism spectrum disorder; Rett syndrome; Fragile X
syndrome; Angelman syndrome; hyperkinetic disorder; mitochondrial
disease; dystonia; a cancer; brain cancer; glioma; breast cancer;
liver cancer; lung cancer; pancreatic cancer; melanoma; ovarian
cancer; gastric cancer; renal cancer; bladder cancer; addiction;
nicotine addiction; smoking; alcohol addiction; drug addiction;
cannabis use disorder; a mental disorder; post-traumatic stress
disorder; anxiety; early psychosis; schizophrenia; a cognitive
disorder; stroke; cardiac ischemia; coronary artery disease;
thromboembolism; myocardial infarction; ischemic related disease; a
gastrointestinal disorder; inflammatory bowel disease; Crohn's
disease; ulcerative colitis; nausea; vomiting; emesis; motion
sickness; chemotherapy induced nausea; chemotherapy induced nausea
vomiting; inflammation; arthritis; rheumatoid arthritis;
osteoarthritis; diabetes; high blood pressure; poor insulin
control; appetite suppression; anorexia; neonatal hypoxic-ischemic
encephalopathy (NHIE); a degenerative skeletal muscle disease; or
Duchenne muscular dystrophy (DMD).
[0064] In a further embodiment of the invention the epilepsy is
Dravet syndrome, Lennox Gastaut syndrome, febrile infection related
epilepsy syndrome (FIRES), Doose syndrome, Sturge Weber syndrome,
CDKL5 mutation; Aicardi syndrome; bilateral polymicrogyria; Dup15q;
SNAP25; benign rolandic epilepsy; juvenile myoclonic epilepsy;
infantile spasm (West syndrome); and Landau-Kleffner syndrome,
refractory epilepsy, juvenile spasms, West syndrome, infantile
spasms, refractory infantile spasms, tuberous sclerosis complex
(TSC); neurogenetic storage disorder, neuronal ceroid
lipofuscinoses (NCL), Batten disease, brain abnormality, atonic,
idiopathic, absence seizure, partial seizure, simple partial
seizure, or complex partial seizure.
BRIEF DESCRIPTION OF THE DRAWINGS
[0065] FIG. 1 depicts the biosynthetic pathway of cannabinoid
production.
[0066] FIG. 2 depicts the different chiral forms of
tetrahydrocannabinol (THC).
[0067] FIG. 3 depicts a UPLC chromatogram detailing trace levels of
cannabinoid in a botanically derived purified CBD preparation.
[0068] FIG. 4 depicts a mass spectrum of cis-THC isolated from a
high-CBD plant.
[0069] FIG. 5 depicts HPLC/DAD traces showing the retention time of
trans-THC (top) and cis-THC (bottom).
[0070] FIG. 6 depicts Spearman rank plots of trans-THC to cis-THC
in CBD material during processing stages.
[0071] FIG. 7 depicts the determination of stereoisomeric form of
cis-THC present in botanically derived purified CBD
preparation.
[0072] FIG. 8 depicts HPLC comparison of botanically derived
purified CBD and synthetic CBD.
[0073] FIG. 9 depicts an autofluorescence spectra of botanically
derived purified CBD and synthetic CBD at 100 mM representative of
the excitation scan from 230 nm to below the emission wavelengths
fixed at 400 or 440 nm.
[0074] FIG. 10 depicts an autofluorescence spectra of botanically
derived purified CBD and synthetic CBD at 100 mM representative of
the emission scan up to 800 nm above the excitation wavelengths
fixed at 326/370 nm for botanically derived purified CBD and
328/334/344 nm for synthetic CBD.
[0075] FIG. 11 depicts an autofluorescence spectra of botanically
derived purified CBD and synthetic CBD at 100 mM representative of
the excitation scan from 230 nm to below the emission wavelengths
fixed at 400 or 440 nm.
[0076] FIG. 12 depicts an autofluorescence spectra of botanically
derived purified CBD and synthetic CBD at 100 mM representative of
the emission scan up to 800 nm above the excitation wavelengths
fixed at 326/370 nm for botanically derived purified CBD and
326/340 nm for synthetic CBD.
[0077] FIG. 13 depicts Experiment 1--sigmoidal curve showing log
dose of CBD versus anticonvulsant activity upon MES test.
[0078] FIG. 14 depicts Experiment 2--sigmoidal curve showing log
dose of CBD versus anticonvulsant activity upon MES test.
[0079] FIG. 15 depicts novel versus familiar exploration times in
rats treated with botanically derived purified CBD.
[0080] FIG. 16 depicts novel versus familiar exploration times in
rats treated with synthetic CBD.
[0081] FIG. 17 depicts novel versus familiar exploration times in
rats treated with botanically derived purified CBD supplemented
with THC at 10 and 20% (w/w).
[0082] FIG. 18 depicts discrimination index in rats treated with
botanically derived purified CBD supplemented with THC at 10 and
20% (w/w).
[0083] FIG. 19 depicts number of line crossings in rats treated
with botanically derived purified CBD supplemented with THC at 10
and 20% (w/w).
DEFINITIONS
[0084] In this application, unless otherwise clear from context,
(i) the term "a" may be understood to mean "at least one"; (ii) the
term "or" may be understood to mean "and/or"; (iii) the terms
"comprising" and "including" may be understood to encompass
itemized components or steps whether presented by themselves or
together with one or more additional components or steps; and (iv)
the terms "about" and "approximately" may be understood to permit
standard variation as would be understood by those of ordinary
skill in the art; and (v) where ranges are provided, endpoints are
included.
[0085] About or Approximately: The terms "about" or
"approximately", as applied to one or more values of interest,
refers to a value that is similar to a stated reference value. In
certain embodiments, the term "about" or "approximately" refers to
a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%,
15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%,
or less in either direction (greater than or less than) of the
stated reference value unless otherwise stated or otherwise evident
from the context.
[0086] Administration: As used herein, the term "administration"
typically refers to the administration of a composition to a
subject or system. Those of ordinary skill in the art will be aware
of a variety of routes that may, in appropriate circumstances, be
utilized for administration to a subject, for example a human. For
example, in some embodiments, administration may be ocular, oral,
parenteral, topical, etc. In some particular embodiments,
administration may be bronchial (e.g., by bronchial instillation),
buccal, dermal (which may be or comprise, for example, one or more
of topical to the dermis, intradermal, interdermal, transdermal,
etc.), enteral, intra-arterial, intradermal, intragastric,
intramedullary, intramuscular, intranasal, intraperitoneal,
intrathecal, intravenous, intraventricular, within a specific organ
(e. g. intrahepatic), mucosal, nasal, oral, rectal, subcutaneous,
sublingual, topical, tracheal (e.g., by intratracheal
instillation), vaginal, vitreal, etc. In some embodiments,
administration may involve dosing that is intermittent (e.g., a
plurality of doses separated in time) and/or periodic (e.g.,
individual doses separated by a common period of time) dosing. In
some embodiments, administration may involve continuous dosing
(e.g., perfusion) for at least a selected period of time.
[0087] Agent: In general, the term "agent", as used herein, may be
used to refer to a compound or entity of any chemical class
including, for example, a polypeptide, nucleic acid, saccharide,
lipid, small molecule, metal, or combination or complex thereof. In
appropriate circumstances, as will be clear from context to those
skilled in the art, the term may be utilized to refer to an entity
that is or comprises a cell or organism, or a fraction, extract, or
component thereof. Alternatively, or additionally, as context will
make clear, the term may be used to refer to a natural product in
that it is found in and/or is obtained from nature. In some
instances, again as will be clear from context, the term may be
used to refer to one or more entities that is man-made in that it
is designed, engineered, and/or produced through action of the hand
of man and/or is not found in nature. In some embodiments, an agent
may be utilized in isolated or pure form; in some embodiments, an
agent may be utilized in crude form. In some embodiments, potential
agents may be provided as collections or libraries, for example
that may be screened to identify or characterize active agents
within them. In some cases, the term "agent" may refer to a
compound or entity that is or comprises a polymer; in some cases,
the term may refer to a compound or entity that comprises one or
more polymeric moieties. In some embodiments, the term "agent" may
refer to a compound or entity that is not a polymer and/or is
substantially free of any polymer and/or of one or more particular
polymeric moieties. In some embodiments, the term may refer to a
compound or entity that lacks or is substantially free of any
polymeric moiety.
[0088] Amelioration: as used herein, refers to the prevention,
reduction or palliation of a state, or improvement of the state of
a subject. Amelioration includes but does not require complete
recovery or complete prevention of a disease, disorder or condition
(e.g., radiation injury).
[0089] Biologically active: as used herein, refers to an observable
biological effect or result achieved by an agent or entity of
interest. For example, in some embodiments, a specific binding
interaction is a biological activity. In some embodiments,
modulation (e.g., induction, enhancement, or inhibition) of a
biological pathway or event is a biological activity. In some
embodiments, presence or extent of a biological activity is
assessed through detection of a direct or indirect product produced
by a biological pathway or event of interest.
[0090] Cancer. The terms "cancer", "malignancy", "neoplasm",
"tumor", and "carcinoma", are used herein to refer to cells that
exhibit relatively abnormal, uncontrolled, and/or autonomous
growth, so that they exhibit an aberrant growth phenotype
characterized by a significant loss of control of cell
proliferation. In some embodiments, a tumor may be or comprise
cells that are precancerous (e.g., benign), malignant,
pre-metastatic, metastatic, and/or non-metastatic. The present
disclosure specifically identifies certain cancers to which its
teachings may be particularly relevant. In some embodiments, a
relevant cancer may be characterized by a solid tumor. In some
embodiments, a relevant cancer may be characterized by a
hematologic tumor. In general, examples of different types of
cancers known in the art include, for example, hematopoietic
cancers including leukemias, lymphomas (Hodgkin's and
non-Hodgkin's), myelomas and myeloproliferative disorders;
sarcomas, melanomas, adenomas, carcinomas of solid tissue, squamous
cell carcinomas of the mouth, throat, larynx, and lung, liver
cancer, genitourinary cancers such as prostate, cervical, bladder,
uterine, and endometrial cancer and renal cell carcinomas, bone
cancer, pancreatic cancer, skin cancer, cutaneous or intraocular
melanoma, cancer of the endocrine system, cancer of the thyroid
gland, cancer of the parathyroid gland, head and neck cancers,
breast cancer, gastro-intestinal cancers and nervous system
cancers, benign lesions such as papilloma's, and the like.
[0091] Carrier as used herein, refers to a diluent, adjuvant,
excipient, or vehicle with which a composition is administered. In
some exemplary embodiments, carriers can include sterile liquids,
such as, for example, water and oils, including oils of petroleum,
animal, vegetable or synthetic origin, such as, for example, peanut
oil, soybean oil, mineral oil, sesame oil and the like. In some
embodiments, carriers are or include one or more solid
components.
[0092] Comparable: As used herein, the term "comparable" refers to
two or more agents, entities, situations, sets of conditions, etc.,
that may not be identical to one another but that are sufficiently
similar to permit comparison there between so that one skilled in
the art will appreciate that conclusions may reasonably be drawn
based on differences or similarities observed. In some embodiments,
comparable sets of conditions, circumstances, individuals, or
populations are characterized by a plurality of substantially
identical features and one or a small number of varied features.
Those of ordinary skill in the art will understand, in context,
what degree of identity is required in any given circumstance for
two or more such agents, entities, situations, sets of conditions,
etc. to be considered comparable. For example, those of ordinary
skill in the art will appreciate that sets of circumstances,
individuals, or populations are comparable to one another when
characterized by a sufficient number and type of substantially
identical features to warrant a reasonable conclusion that
differences in results obtained or phenomena observed under or with
different sets of circumstances, individuals, or populations are
caused by or indicative of the variation in those features that are
varied.
[0093] Composition: Those skilled in the art will appreciate that
the term "composition" may be used to refer to a discrete physical
entity that comprises one or more specified components. In general,
unless otherwise specified, a composition may be of any form--e.g.,
gas, gel, liquid, solid, etc.
[0094] Comprising: A composition or method described herein as
"comprising" one or more named elements or steps is open-ended,
meaning that the named elements or steps are essential, but other
elements or steps may be added within the scope of the composition
or method. To avoid prolixity, it is also understood that any
composition or method described as "comprising" (or which
"comprises") one or more named elements or steps also describes the
corresponding, more limited composition or method "consisting
essentially of" (or which "consists essentially of") the same named
elements or steps, meaning that the composition or method includes
the named essential elements or steps and may also include
additional elements or steps that do not materially affect the
basic and novel characteristic(s) of the composition or method. It
is also understood that any composition or method described herein
as "comprising" or "consisting essentially of" one or more named
elements or steps also describes the corresponding, more limited,
and closed-ended composition or method "consisting of" (or
"consists of") the named elements or steps to the exclusion of any
other unnamed element or step. In any composition or method
disclosed herein, known or disclosed equivalents of any named
essential element or step may be substituted for that element or
step.
[0095] Determine: Many methodologies described herein include a
step of "determining". Those of ordinary skill in the art, reading
the present specification, will appreciate that such "determining"
can utilize or be accomplished through use of any of a variety of
techniques available to those skilled in the art, including for
example specific techniques explicitly referred to herein. In some
embodiments, determining involves manipulation of a physical
sample. In some embodiments, determining involves consideration
and/or manipulation of data or information, for example utilizing a
computer or other processing unit adapted to perform a relevant
analysis. In some embodiments, determining involves receiving
relevant information and/or materials from a source. In some
embodiments, determining involves comparing one or more features of
a sample or entity to a comparable reference.
[0096] Dosage form or unit dosage form: Those skilled in the art
will appreciate that the term "dosage form" may be used to refer to
a physically discrete unit of an active agent (e.g., a therapeutic
or diagnostic agent) for administration to a subject. Typically,
each such unit contains a predetermined quantity of active agent.
In some embodiments, such quantity is a unit dosage amount (or a
whole fraction thereof) appropriate for administration in
accordance with a dosing regimen that has been determined to
correlate with a desired or beneficial outcome when administered to
a relevant population (i.e., with a therapeutic dosing regimen).
Those of ordinary skill in the art appreciate that the total amount
of a therapeutic composition or agent administered to a particular
subject is determined by one or more attending physicians and may
involve administration of multiple dosage forms.
[0097] Dosing regimen: Those skilled in the art will appreciate
that the term "dosing regimen" may be used to refer to a set of
unit doses (typically more than one) that are administered
individually to a subject, typically separated by periods of time.
In some embodiments, a given therapeutic agent has a recommended
dosing regimen, which may involve one or more doses. In some
embodiments, a dosing regimen comprises a plurality of doses each
of which is separated in time from other doses. In some
embodiments, individual doses are separated from one another by a
time period of the same length; in some embodiments, a dosing
regimen comprises a plurality of doses and at least two different
time periods separating individual doses. In some embodiments, all
doses within a dosing regimen are of the same unit dose amount. In
some embodiments, different doses within a dosing regimen are of
different amounts. In some embodiments, a dosing regimen comprises
a first dose in a first dose amount, followed by one or more
additional doses in a second dose amount different from the first
dose amount. In some embodiments, a dosing regimen comprises a
first dose in a first dose amount, followed by one or more
additional doses in a second dose amount same as the first dose
amount. In some embodiments, a dosing regimen is correlated with a
desired or beneficial outcome when administered across a relevant
population (i.e., is a therapeutic dosing regimen).
[0098] Excipient: as used herein, refers to a non-therapeutic agent
that may be included in a pharmaceutical composition, for example
to provide or contribute to a desired consistency or stabilizing
effect. Suitable pharmaceutical excipients include, for example,
starch, glucose, lactose, sucrose, gelatin, malt, rice, flour,
chalk, silica gel, sodium stearate, glycerol monostearate, talc,
sodium chloride, dried skim milk, glycerol, propylene, glycol,
water, ethanol and the like.
[0099] Improve, increase, inhibit or reduce: As used herein, these
terms, or grammatically comparable comparative terms, indicate
values that are relative to a comparable reference measurement. For
example, in some embodiments, an assessed value achieved with an
agent of interest may be "improved" relative to that obtained with
a comparable reference agent. Alternatively or additionally, in
some embodiments, an assessed value achieved in a subject or system
of interest may be "improved" relative to that obtained in the same
subject or system under different conditions (e.g., prior to or
after an event such as administration of an agent of interest), or
in a different, comparable subject (e.g., in a comparable subject
or system that differs from the subject or system of interest in
presence of one or more indicators of a particular disease,
disorder or condition of interest, or in prior exposure to a
condition or agent, etc.). In some embodiments, comparative terms
refer to statistically relevant differences (e.g., that are of a
prevalence and/or magnitude sufficient to achieve statistical
relevance). Those skilled in the art will be aware, or will readily
be able to determine, in a given context, a degree and/or
prevalence of difference that is required or sufficient to achieve
such statistical significance.
[0100] In vitro: The term "in vitro" as used herein refers to
events that occur in an artificial environment, e.g., in a test
tube or reaction vessel, in cell culture, etc., rather than within
a multi-cellular organism.
[0101] In vivo: as used herein refers to events that occur within a
multi-cellular organism, such as a human and a non-human animal. In
the context of cell-based systems, the term may be used to refer to
events that occur within a living cell (as opposed to, for example,
in vitro systems).
[0102] Isolated: as used herein, refers to a substance and/or
entity that has been (1) separated from at least some of the
components with which it was associated when initially produced
(whether in nature and/or in an experimental setting), and/or (2)
designed, produced, prepared, and/or manufactured by the hand of
man. In some embodiments, isolated agents are about 80%, about 85%,
about 90%, about 91%, about 92%, about 93%, about 94%, about 95%,
about 96%, about 97%, about 98%, about 99%, or more than about 99%
pure. As used herein, a substance is "pure" if it is substantially
free of other components.
[0103] Isomer: As is known in the art, many chemical entities (in
particular many organic molecules and/or many small molecules) can
exist in a variety of structural (e.g., geometric/conformational)
and/or optical isomeric forms. For example, any chiral center can
exist in R and S configurations; double bonds can exist in Z and E
conformational isomers, certain structural elements can adopt two
or more tautomeric forms, etc. In some embodiments, as will be
clear to those skilled in the art from context, depiction of or
reference to a particular compound structure herein may represent
all structural and/or optical isomers thereof. In some embodiments,
as will be clear to those skilled in the art from context,
depiction of or reference to a particular compound structure herein
is intended to encompass only the depicted or referenced isomeric
form. In some embodiments, compositions including a chemical entity
that can exist in a variety of isomeric forms include a plurality
of such forms; in some embodiments such compositions include only a
single form. For example, in some embodiments, compositions
including a chemical entity that can exist as a variety of optical
isomers (e.g., stereoisomers, diastereomers, etc.) include a
racemic population of such optical isomers; in some embodiments
such compositions include only a single optical isomer and/or
include a plurality of optical isomers that together retain optical
activity. Where there exists two or more isomers within a
composition they may exist as a mixture with various ratios.
[0104] Mixture: The phrase "mixture" describes a combination of two
or more different compounds or agents which occur within the same
composition.
[0105] Moiety: Those skilled in the art will appreciate that a
"moiety" is a defined chemical group or entity with a particular
structure and/or or activity, as described herein.
[0106] Oral: The phrases "oral administration" and "administered
orally" as used herein have their art-understood meaning referring
to administration by mouth of a compound or composition.
[0107] Parenteral: The phrases "parenteral administration" and
"administered parenterally" as used herein have their
art-understood meaning referring to modes of administration other
than enteral and topical administration, usually by injection, and
include, without limitation, intravenous, intramuscular,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticulare, subcapsular,
subarachnoid, intraspinal, and intrasternal injection and
infusion.
[0108] Patient: As used herein, the term "patient" refers to any
organism to which a provided composition is or may be administered,
e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or
therapeutic purposes. Typical patients include animals (e.g.,
mammals such as mice, rats, rabbits, non-human primates, and/or
humans). In some embodiments, a patient is a human. In some
embodiments, a patient is suffering from or susceptible to one or
more disorders or conditions. In some embodiments, a patient
displays one or more symptoms of a disorder or condition. In some
embodiments, a patient has been diagnosed with one or more
disorders or conditions. In some embodiments, the disorder or
condition is or includes cancer, or presence of one or more tumors.
In some embodiments, the patient is receiving or has received
certain therapy to diagnose and/or to treat a disease, disorder, or
condition.
[0109] Pharmaceutical composition: As used herein, the term
"pharmaceutical composition" refers to an active agent, formulated
together with one or more pharmaceutically acceptable carriers. In
some embodiments, active agent is present in unit dose amount
appropriate for administration in a therapeutic regimen that shows
a statistically significant probability of achieving a
predetermined therapeutic effect when administered to a relevant
population. In some embodiments, pharmaceutical compositions may be
specially formulated for administration in solid or liquid form,
including those adapted for the following: oral administration, for
example, drenches (aqueous or non-aqueous solutions or
suspensions), tablets, e.g., those targeted for buccal, sublingual,
and systemic absorption, boluses, powders, granules, pastes for
application to the tongue; parenteral administration, for example,
by subcutaneous, intramuscular, intravenous or epidural injection
as, for example, a sterile solution or suspension, or
sustained-release formulation; topical application, for example, as
a cream, ointment, or a controlled-release patch or spray applied
to the skin, lungs, or oral cavity; intravaginally or
intrarectally, for example, as a pessary, cream, or foam;
sublingually; ocularly; transdermally; or nasally, pulmonary, and
to other mucosal surfaces.
[0110] Pharmaceutically acceptable: As used herein, the phrase
"pharmaceutically acceptable" refers to those compounds, materials,
compositions, and/or dosage forms which are, within the scope of
sound medical judgment, suitable for use in contact with the
tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other problem or complication,
commensurate with a reasonable benefit/risk ratio.
[0111] Pharmaceutically acceptable carrier: As used herein, the
term "pharmaceutically acceptable carrier" means a
pharmaceutically-acceptable material, composition or vehicle, such
as a liquid or solid filler, diluent, excipient, or solvent
encapsulating material, involved in carrying or transporting the
subject compound from one organ, or portion of the body, to another
organ, or portion of the body. Each carrier must be "acceptable" in
the sense of being compatible with the other ingredients of the
formulation and not injurious to the patient. Some examples of
materials which can serve as pharmaceutically-acceptable carriers
include: sugars, such as lactose, glucose and sucrose; starches,
such as corn starch and potato starch; cellulose, and its
derivatives, such as sodium carboxymethyl cellulose, ethyl
cellulose and cellulose acetate; powdered tragacanth; malt;
gelatin; talc; excipients, such as cocoa butter and suppository
waxes; oils, such as peanut oil, cottonseed oil, safflower oil,
sesame oil, olive oil, corn oil and soybean oil; glycols, such as
propylene glycol; polyols, such as glycerin, sorbitol, mannitol and
polyethylene glycol; esters, such as ethyl oleate and ethyl
laurate; agar; buffering agents, such as magnesium hydroxide and
aluminum hydroxide; alginic acid; pyrogen-free water; isotonic
saline; Ringer's solution; ethyl alcohol; pH buffered solutions;
polyesters, polycarbonates and/or polyanhydrides; and other
non-toxic compatible substances employed in pharmaceutical
formulations.
[0112] Predetermined: By predetermined is meant deliberately
selected, for example as opposed to randomly occurring or
achieved.
[0113] Prevent or prevention: as used herein when used in
connection with the occurrence of a disease, disorder, and/or
condition, refers to reducing the risk of developing the disease,
disorder and/or condition and/or to delaying onset of one or more
characteristics or symptoms of the disease, disorder or condition.
Prevention may be considered complete when onset of a disease,
disorder or condition has been delayed for a predefined period of
time.
[0114] Predominantly present: The term "predominantly present", as
used herein, refers to the quantity of an entity (e.g., a specific
cannabinoid or isomer thereof) in a preparation or composition. For
example, a cannabinoid may be predominantly present if it is at
least about 50%, about 55%, about 60%, about 65%, about 70%, about
75%, about 80%, about 85%, about 90%, about 95%, about 96%, about
97%, about 98%, about 99%, or about 100% of the total cannabinoid
in the preparation or composition.
[0115] Prevention: The term "prevention", as used herein, refers to
a delay of onset, and/or reduction in frequency and/or severity of
one or more symptoms of a particular disease, disorder or
condition. In some embodiments, prevention is assessed on a
population basis such that an agent is considered to "prevent" a
particular disease, disorder or condition if a statistically
significant decrease in the development, frequency, and/or
intensity of one or more symptoms of the disease, disorder or
condition is observed in a population susceptible to the disease,
disorder, or condition. Prevention may be considered complete when
onset of a disease, disorder or condition has been delayed for a
predefined period of time.
[0116] Pure: As used herein, an agent or entity is "pure" if it is
substantially free of other components. For example, a preparation
that contains more than about 90% of a particular agent or entity
is typically considered to be a pure preparation. In some
embodiments, an agent or entity is at least 91%, at least 92%, at
least 93%, at least 94%, at least 95%, at least 96%, at least 97%,
at least 98%, or at least 99% pure.
[0117] Reference: As used herein describes a standard or control
relative to which a comparison is performed. For example, in some
embodiments, an agent, animal, individual, population, sample,
sequence or value of interest is compared with a reference or
control agent, animal, individual, population, sample, sequence or
value. In some embodiments, a reference composition may comprise
one or more synthetic cannabinoids. In some embodiments a reference
composition may contain different types of cannabinoids, different
isomeric forms of cannabinoids, different distribution of
cannabinoids, different quantities of cannabinoids, etc. as
compared to a test composition. In some embodiments, a reference or
control is tested and/or determined substantially simultaneously
with the testing or determination of interest. In some embodiments,
a reference or control is a historical reference or control,
optionally embodied in a tangible medium. Typically, as would be
understood by those skilled in the art, a reference or control is
determined or characterized under comparable conditions or
circumstances to those under assessment. Those skilled in the art
will appreciate when sufficient similarities are present to justify
reliance on and/or comparison to a particular possible reference or
control.
[0118] Response: As used herein, a response to treatment may refer
to any beneficial alteration in a subject's condition that occurs
as a result of or correlates with treatment. Such alteration may
include stabilization of the condition (e.g., prevention of
deterioration that would have taken place in the absence of the
treatment), amelioration of symptoms of the condition, and/or
improvement in the prospects for cure of the condition, etc.
Response may be measured according to a wide variety of criteria,
including clinical criteria and objective criteria. Techniques for
assessing response include, but are not limited to, clinical
examination, positron emission tomatography, chest X-ray CT scan,
MRI, ultrasound, endoscopy, laparoscopy, presence or level of tumor
markers in a sample obtained from a subject, cytology, and/or
histology. The exact response criteria can be selected in any
appropriate manner, provided that when comparing groups of cells or
subjects, the groups to be compared are assessed based on the same
or comparable criteria for determining response rate. One of
ordinary skill in the art will be able to select appropriate
criteria.
[0119] Solid form: As is known in the art, many chemical entities
(in particular many organic molecules and/or many small molecules)
can adopt a variety of different solid forms such as, for example,
amorphous forms and/or crystalline forms (e.g., polymorphs,
hydrates, solvates, etc.). In some embodiments, such entities may
be utilized as a single such form (e.g., as a pure preparation of a
single polymorph). In some embodiments, such entities may be
utilized as a mixture of such forms.
[0120] Subject: As used herein, the term "subject" refers an
organism, typically a mammal (e.g., a human, in some embodiments
including prenatal human forms). In some embodiments, a subject
refers to any organism (e.g., mammals such as mice, rats, rabbits,
non-human primates, and humans; insects; worms; etc.) and plants to
which a provided compound or composition is administered in
accordance with the present disclosure e.g., for experimental,
diagnostic, prophylactic, and/or therapeutic purposes. In some
embodiments, a subject is suffering from a relevant disease,
disorder or condition. In some embodiments, a subject is
susceptible to a disease, disorder, or condition. In some
embodiments, a subject displays one or more symptoms or
characteristics of a disease, disorder or condition. In some
embodiments, a subject does not display any symptom or
characteristic of a disease, disorder, or condition. In some
embodiments, a subject is someone with one or more features
characteristic of susceptibility to or risk of a disease, disorder,
or condition. In some embodiments, a subject is a patient. In some
embodiments, a subject is an individual to whom diagnosis and/or
therapy is and/or has been administered.
[0121] Substantially: As used herein, the term "substantially"
refers to the qualitative condition of exhibiting total or
near-total extent or degree of a characteristic or property of
interest. One of ordinary skill in the biological arts will
understand that biological and chemical phenomena rarely, if ever,
go to completion and/or proceed to completeness or achieve or avoid
an absolute result. The term "substantially" is therefore used
herein to capture the potential lack of completeness inherent in
many biological and chemical phenomena.
[0122] Suffering from: An individual who is "suffering from" a
disease, disorder, and/or condition has been diagnosed with and/or
displays one or more symptoms of a disease, disorder, and/or
condition.
[0123] Susceptible to: An individual who is "susceptible to" a
disease, disorder, and/or condition is one who has a higher risk of
developing the disease, disorder, and/or condition than does a
member of the general public. In some embodiments, an individual
who is susceptible to a disease, disorder and/or condition may not
have been diagnosed with the disease, disorder, and/or condition.
In some embodiments, an individual who is susceptible to a disease,
disorder, and/or condition may exhibit symptoms of the disease,
disorder, and/or condition. In some embodiments, an individual who
is susceptible to a disease, disorder, and/or condition may not
exhibit symptoms of the disease, disorder, and/or condition. In
some embodiments, an individual who is susceptible to a disease,
disorder, and/or condition will develop the disease, disorder,
and/or condition. In some embodiments, an individual who is
susceptible to a disease, disorder, and/or condition will not
develop the disease, disorder, and/or condition.
[0124] Symptoms are reduced: According to the present invention,
"symptoms are reduced" when one or more symptoms of a particular
disease, disorder or condition is reduced in magnitude (e.g.,
intensity, severity, etc.) and/or frequency. For purposes of
clarity, a delay in the onset of a particular symptom is considered
one form of reducing the frequency of that symptom.
[0125] Systemic: The phrases "systemic administration,"
"administered systemically," "peripheral administration," and
"administered peripherally" as used herein have their
art-understood meaning referring to administration of a compound or
composition such that it enters the recipient's system.
[0126] Therapeutic agent: As used herein, the phrase "therapeutic
agent" in general refers to any agent that elicits a desired
pharmacological effect when administered to an organism. In some
embodiments, an agent is considered to be a therapeutic agent if it
demonstrates a statistically significant effect across an
appropriate population. In some embodiments, the appropriate
population may be a population of model organisms. In some
embodiments, an appropriate population may be defined by various
criteria, such as a certain age group, gender, genetic background,
preexisting clinical conditions, etc. In some embodiments, a
therapeutic agent is a substance that can be used to alleviate,
ameliorate, relieve, inhibit, prevent, delay onset of, reduce
severity of, and/or reduce incidence of one or more symptoms or
features of a disease, disorder, and/or condition. In some
embodiments, a "therapeutic agent" is an agent that has been or is
required to be approved by a government agency before it can be
marketed for administration to humans. In some embodiments, a
"therapeutic agent" is an agent for which a medical prescription is
required for administration to humans.
[0127] Therapeutic regimen: A "therapeutic regimen", as that term
is used herein, refers to a dosing regimen whose administration
across a relevant population may be correlated with a desired or
beneficial therapeutic outcome.
[0128] Therapeutically effective amount: As used herein, the term
"therapeutically effective amount" means an amount of a substance
(e.g., a therapeutic agent, active ingredient, preparation,
composition, and/or formulation) that elicits a desired a desired
effect (e.g., a desired biological, clinical, or pharmacological
effect or response) when administered as part of a therapeutic
regimen. In some embodiments, a therapeutically effective amount of
a substance is an amount that is sufficient, when administered to a
subject suffering from or susceptible to a disease, disorder,
and/or condition, to treat, diagnose, prevent, reduce the severity
of, stabilize one or more characteristics of, and/or delay the
onset of the disease, disorder, and/or condition. In some
embodiments, the term refers to an amount sufficient to produce the
effect in at least a significant percentage (e.g., at least about
25%, about 30%, about 40%, about 50%, about 60%, about 70%, about
80%, about 90%, about 95%, or more) of a population that is
suffering from and/or susceptible to a disease, disorder, and/or
condition. As will be appreciated by those of ordinary skill in
this art, the effective amount of a substance may vary depending on
such factors as the desired biological endpoint, the substance to
be delivered, the target cell or tissue, etc. For example, the
effective amount of compound in a formulation to treat a disease,
disorder, and/or condition is the amount that alleviates,
ameliorates, relieves, inhibits, prevents, delays onset of, reduces
severity of and/or reduces incidence of one or more symptoms or
features of the disease, disorder, and/or condition. In some
embodiments, a therapeutically effective amount is administered in
a single dose; in some embodiments, multiple unit doses are
required to deliver a therapeutically effective amount.
[0129] Treat: As used herein, the term "treat," "treatment," or
"treating" refers to any method used to partially or completely
alleviate, ameliorate, relieve, inhibit, prevent, delay onset of,
reduce severity of, and/or reduce incidence of one or more symptoms
or features of a disease, disorder, and/or condition. In some
embodiments, treatment refers to administration of a therapy that
partially or completely alleviates, ameliorates, relives, inhibits,
delays onset of, reduces severity of, and/or reduces incidence of
one or more symptoms, features, and/or causes of a particular
disease, disorder, and/or condition. Treatment may be administered
to a subject who does not exhibit signs of a disease, disorder,
and/or condition. In some embodiments, treatment may be
administered to a subject who exhibits only early signs of the
disease, disorder, and/or condition, for example for the purpose of
decreasing the risk of developing pathology associated with the
disease, disorder, and/or condition.
[0130] Unit dose: The expression "unit dose" as used herein refers
to an amount administered as a single dose and/or in a physically
discrete unit of a pharmaceutical composition. In many embodiments,
a unit dose contains a predetermined quantity of an active agent.
In some embodiments, a unit dose contains an entire single dose of
the agent. In some embodiments, more than one unit dose is
administered to achieve a total single dose. In some embodiments,
administration of multiple unit doses is required, or expected to
be required, in order to achieve an intended effect. A unit dose
may be, for example, a volume of liquid (e.g., an acceptable
carrier) containing a predetermined quantity of one or more
therapeutic agents, a predetermined amount of one or more
therapeutic agents in solid form, a sustained release formulation
or drug delivery device containing a predetermined amount of one or
more therapeutic agents, etc. It will be appreciated that a unit
dose may be present in a formulation that includes any of a variety
of components in addition to the therapeutic agent(s). For example,
acceptable carriers (e.g., pharmaceutically acceptable carriers),
diluents, stabilizers, buffers, preservatives, etc., may be
included as described infra. It will be appreciated by those
skilled in the art, in many embodiments, a total appropriate daily
dosage of a particular therapeutic agent may comprise a portion, or
a plurality, of unit doses, and may be decided, for example, by the
attending physician within the scope of sound medical judgment. In
some embodiments, the specific effective dose level for any
particular subject or organism may depend upon a variety of factors
including the disorder being treated and the severity of the
disorder; activity of specific active compound employed; specific
composition employed; age, body weight, general health, sex and
diet of the subject; time of administration, and rate of excretion
of the specific active compound employed; duration of the
treatment; drugs and/or additional therapies used in combination or
coincidental with specific compound(s) employed, and like factors
well known in the medical arts.
DETAILED DESCRIPTION
[0131] Cannabinoids are natural and synthetic compounds
structurally or pharmacologically related to the constituents of
the Cannabis plant or to the endogenous agonists (endocannabinoids)
of the cannabinoid receptors CB1 or CB2. The only way in nature in
which these compounds are produced is by the Cannabis plant.
Cannabis is a genus of flowering plants in the family Cannabaceae,
comprising the species Cannabis sativa, Cannabis indica, and
Cannabis ruderalis (sometimes considered as part of Cannabis
sativa).
[0132] Cannabis plants comprise a highly complex mixture of
compounds. At least 568 unique molecules have been identified.
Among these compounds are cannabinoids, terpenoids, sugars, fatty
acids, flavonoids, other hydrocarbons, nitrogenous compounds, and
amino acids.
[0133] Cannabinoids exert their physiological effects through a
variety of receptors including, but not limited to, adrenergic
receptors, cannabinoid receptors (CB1 and CB2), GPR55, GPR3, or
GPR5. The principle cannabinoids present in Cannabis plants are
cannabinoid acids tetrahydrocannabinolic acid (THCA) and
cannabidiolic acid (CBDA) with small amounts of their respective
neutral (decarboxylated) cannabinoids. In addition, cannabis may
contain lower levels of other minor cannabinoids. "Chemical
composition, pharmacological profiling, and complete physiological
effects of these medicinal plants, and more importantly the
extracts from cannabis, remain to be fully understood." Lewis, M.
M. et al., ACS Omega, 2, 6091-6103 (2017). FIG. 1 depicts an
exemplary schematic of the biosynthetic pathways of certain
phytocannabinoids.
[0134] Crude extracts from Cannabis plants containing CBD have been
used by patients suffering from diseases and disorders. However,
such crude products are unsuitable for use in pharmaceutical
formulations. Those seeking to prepare more consistent CBD
preparations for use in treating diseases or disorders have made a
concerted effort to either prepare CBD synthetically or attempt to
remove all compounds other than CBD, particularly psychoactive
compounds such as THC, from plant derived cannabinoids.
[0135] The present invention encompasses the surprising discovery
that a botanically derived purified CBD preparation, comprising one
or more additional cannabinoids, and suitable for pharmaceutical
use, exhibits enhanced therapeutic efficacy when compared to prior
CBD preparations which differ from the composition disclosed
herein.
[0136] These preparations differ either by being purified to the
extent that no other impurities exist or being produced
synthetically thereby comprising no additional cannabinoids that
would be produced by nature or further differ by being an
unpurified plant extract which extract comprises some or all of the
cannabinoids and non-cannabinoid compounds that are co-produced by
the plant and co-extracted in the preparation of the extract. In
some embodiments botanically derived purified CBD preparation of
the present invention may be administered in a lower dose of CBD
than a synthetic or completely pure preparation of CBD.
Cannabinoids
[0137] As stated, cannabinoids are a class of compounds which may
be derived naturally from the Cannabis plant or produced
synthetically via chemical synthesis.
[0138] More than 100 different cannabinoids produced by Cannabis
have been identified. These cannabinoids can be split into
different groups as follows: phytocannabinoids; endocannabinoids
and synthetic cannabinoids (which may be novel cannabinoids or
synthetically produced versions of phytocannabinoids or
endocannabinoids).
[0139] Phytocannabinoids are cannabinoids that originate from
nature and can be found in the Cannabis plant. Phytocannabinoids
can be isolated from plants to produce a highly purified extract.
Phytocannabinoids may be obtained as either the neutral
(decarboxylated form) or the carboxylic acid form depending on the
method used to extract the cannabinoids from plant material. For
example, it is known that heating the carboxylic acid form will
cause most of the carboxylic acid form to decarboxylate into the
neutral form. Phytocannabinoids can only be produced from plants,
however versions of phytocannabinoids may be produced synthetically
via chemical synthesis.
[0140] Endocannabinoids are endogenous lipid-based retrograde
neurotransmitters that bind to cannabinoid receptors, and
cannabinoid receptor proteins that are expressed throughout the
mammalian central nervous system (including the brain) and
peripheral nervous system. The endocannabinoid system is involved
in regulating a variety of physiological and cognitive processes
including fertility, pregnancy, during pre- and postnatal
development, appetite, pain-sensation, mood, and memory, and in
mediating the pharmacological effects of Cannabis.
[0141] "Synthetic cannabinoids" are compounds that have a
cannabinoid-like structure and are manufactured using chemical
means rather than by the plant.
[0142] Certain cannabinoids are described in more detail below.
Although little is known about these cannabinoids, CBD preparations
and compositions described herein which comprise one or more of
these components show surprising efficacy, particularly when
compared with pure and/or synthetic CBD compositions.
Cannabidiol (CBD)
[0143] CBD is a major cannabinoid constituent of Cannabis species,
such as the hemp plant (Cannabis sativa). Unlike other
cannabinoids, such as THC, CBD does not bind CB1 or CB2, or its
binding to the receptors is negligible in terms of inducing a
pharmacological effect. Thus, CBD does not cause the central or
peripheral nervous system effects mediated by the CB1 or CB2
receptors. CBD has little or no psychotropic (cannabimimetic)
activity and its molecular structure and properties are
substantially different from those of other cannabinoids.
[0144] CBD administration has been the subject of research in an
attempt to provide an alternative treatment for various diseases
and disorders which may respond to such treatment.
[0145] In some embodiments CBD is isolated from a Cannabis plant.
In some embodiments CBD is prepared synthetically. In some
embodiments, CBD is present as (-)-trans-CBD.
Tetrahydrocannabinol (THC)
[0146] THC is the principal psychoactive constituent of
Cannabis.
[0147] The THC molecule may exist as four distinct chiral forms as
shown in FIG. 2. THC has 2 stereocenters which in turn enable the
existence of 4 stereoisomers: (+)-trans-THC; (-)-trans-THC;
(+)-cis-THC and (-)-cis-THC. THC commonly occurs in nature as the
(-)-trans-THC isomer (Hollister, 1970)
[0148] The THC molecule mostly occurs as
(-)-trans-.DELTA..sup.9-tetrahydrocannabinol, however the
(-)-trans-.DELTA..sup.8-tetrahydrocannabinol homolog is also known
to exist. The skilled person will appreciate that reference to the
compound THC may refer to either the .DELTA..sup.8 or the
.DELTA..sup.9 homolog.
[0149] In some embodiments THC is isolated from a Cannabis plant.
In some embodiments THC is prepared synthetically. In some
embodiments, THC is present as (-)-trans-THC. In some embodiments,
THC is present as (-)-cis-THC. In some embodiments, THC is present
as (+)-trans-THC. In some embodiments, THC is present as
(+)-cis-THC.
[0150] In some embodiments the THC is present as a mixture of
isomers. In some embodiments the mixture will comprise one or more
of (+)-trans-THC, (-)-trans-THC, (+)-cis-THC and (-)-cis-THC.
Cannabidivarin (CBDV)
[0151] CBDV is a homolog of CBD, with the side-chain shortened by
two methylene bridges. In some embodiments CBDV is isolated from a
Cannabis plant. In some embodiments CBDV is prepared synthetically.
In some embodiments, CBDV is present as (-)-trans-CBDV.
Cannabidiol-C1 (CBD-C1)
[0152] In some embodiments CBD-C1 is isolated from a Cannabis
plant. In some embodiments CBD-C1 is prepared synthetically. In
some embodiments, CBD-C1 is present as (-)-trans-CBD-C1.
Cannabidiol-C4 (CBD-C4)
[0153] In some embodiments CBD-C4 is isolated from a Cannabis
plant. In some embodiments CBD-C4 is prepared synthetically. In
some embodiments, CBD-C4 is present as (-)-trans-CBD-C4.
CBD Preparations
[0154] The present disclosure provides certain CBD preparations,
characterized by chemical components and/or functional properties
that distinguish them from prior CBD compositions.
[0155] In some embodiments, a CBD preparation comprises about 90%,
about 91%, about 92%, about 93%, about 94%, about 95%, about 96%,
about 97%, about 98%, about 99%, or more than about 99% of the
other components with which they were initially associated.
Preferably the CBD preparation comprises at least 98% CBD based on
total amount of cannabinoid in the preparation.
[0156] In some embodiments, the CBD comprises (-)-trans-CBD
isoform.
[0157] In some embodiments, the CBD preparation further comprises
tetrahydrocannabinol (THC). In some embodiments, a CBD preparation
comprises up to about 1%, about 2%, about 3%, about 4%, or about 5%
THC based on total amount of cannabinoid in the preparation. In
some embodiments, a CBD preparation comprises not more than 0.15%
THC based on total amount of cannabinoid in the preparation. In
some embodiments, a CBD preparation comprises about 0.01% to about
0.1% THC based on total amount of cannabinoid in the preparation.
In some embodiments, a CBD preparation comprises about 0.02% to
about 0.05% THC based on total amount of cannabinoid in the
preparation. In some embodiments, a CBD preparation comprises at
least about 0.1% THC based on total amount of cannabinoid in the
preparation. In some embodiments, a CBD preparation comprises at
least about 0.02% THC based on total amount of cannabinoid in the
preparation. In some embodiments, the THC comprises
.DELTA..sup.9-THC.
[0158] In some embodiments, the THC is present as a mixture of
different isomers. In some embodiments, the THC comprises trans-THC
and cis-THC. In some embodiments, the trans-THC and cis-THC are
present at a ratio of about 5:1 (trans-THC:cis-THC). In some
embodiments, the trans-THC and cis-THC are present at a ratio of
about 3.5:1 (trans-THC:cis-THC). In some embodiments, the trans-THC
and cis-THC are present at a ratio of about 2:1
(trans-THC:cis-THC). In some embodiments, the trans-THC and cis-THC
are present at a ratio of about 1:1 (trans-THC:cis-THC). In some
embodiments, the trans-THC and cis-THC are present at a ratio of
about 0.8:1 (trans-THC:cis-THC).
[0159] In some embodiments the cis-THC is present as a mixture of
(-)-cis-THC and (+)-cis-THC. In some embodiments, the (-)-cis-THC
and (+)-cis-THC are present at a ratio of about 20:1 to 1:20
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 15:1 to 1:15
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 10:1 to 1:10
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 9:1 to 1:9
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 5:1 to 1:5
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 3:1 to 1:3
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 2:1 to 1:2
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 1:1
((-)-cis-THC:(+)-cis-THC). In some embodiments, the (-)-cis-THC and
(+)-cis-THC are present at a ratio of about 9:1
((-)-cis-THC:(+)-cis-THC).
[0160] In some embodiments, a CBD preparation comprises one or more
cannabinoids other than THC. In some embodiments, a CBD preparation
comprises no more than 2% cannabinoids other than CBD based on
total amount of cannabinoid in the preparation.
[0161] In some embodiments, a CBD preparation comprises
cannabidivarin (CBDV). In some embodiments, the CBDV comprises the
(-)-trans-CBDV isoform. In some embodiments, a CBD preparation
comprises about 0.2% to about 0.8% CBDV based on total amount of
cannabinoid in the preparation.
[0162] In some embodiments, a CBD preparation comprises CBD-C4
(CBD-C4). In some embodiments, the CBD-C4 comprises
(-)-trans-CBD-C4 isoform. In some embodiments, a CBD preparation
comprises about 0.3% to about 0.4% CBD-C4 based on total amount of
cannabinoid in the preparation.
[0163] In some embodiments, a CBD preparation comprises CBD-C1
(CBD-C1). In some embodiments, the CBD-C1 comprises
(-)-trans-CBD-C1 isoform. In some embodiments, a CBD preparation
comprises about 0.1% to about 0.15% CBD-C1 based on total amount of
cannabinoid in the preparation.
[0164] In some embodiments, at least a portion of at least one of
the cannabinoids present in a CBD preparation is isolated from
Cannabis plant material. In some embodiments, at least a portion of
the CBD present in a CBD preparation is isolated from Cannabis
plant material. In some embodiments, at least a portion of the THC
present in a CBD preparation is isolated from Cannabis plant
material. In some embodiments, substantially all of at least one of
the cannabinoids present in a CBD preparation is isolated from
Cannabis plant material. In some embodiments, substantially all the
CBD present in a CBD preparation is isolated from Cannabis plant
material. In some embodiments, substantially all the THC present in
a CBD preparation is isolated from Cannabis plant material. In some
embodiments, substantially all of the cannabinoids present in a CBD
preparation are isolated from Cannabis plant material. In some
embodiments, the Cannabis plant material is from a Cannabis sativa,
Cannabis indica, or Cannabis ruderalis plant. In some embodiments,
the Cannabis plant is a high-CBD containing Cannabis chemotype. In
some embodiments, the Cannabis plant is a high-CBD containing
Cannabis chemotype of Cannabis sativa L. In some embodiments, the
Cannabis plant material comprises about 5% to about 20% CBD based
on total amount of cannabinoid in the preparation. In some
embodiments, the Cannabis plant material comprises about 10% to
about 15% CBD based on total amount of cannabinoid in the
preparation. In some embodiments, the Cannabis plant material
comprises trans-THC and cis-THC are present at a ratio of about
3.5:1 (trans-THC:cis-THC). In some embodiments, the Cannabis plant
material comprises trans-THC and cis-THC are present at a ratio of
about 0.8:1 (trans-THC:cis-THC).
Methods of Making CBD Preparations
[0165] In the context of this application a "botanical drug
substance" is an extract derived from Cannabis plant material,
which extract fulfils the definition of "botanical drug substance"
provided in the Guidance for Industry Botanical Drug Products Draft
Guidance, August 2000, US Department of Health and Human Services,
Food and Drug Administration Center for Drug Evaluation and
Research of: "A drug substance derived from one or more plants,
algae, or macroscopic fungi. It is prepared from botanical raw
materials by one or more of the following processes: pulverization,
decoction, expression, aqueous extraction, ethanolic extraction, or
other similar processes."
[0166] "Plant material" is defined as a plant or plant material
(e.g. bark, wood, leaves, stems, roots, flowers, fruits, seeds,
berries or parts thereof) as well as exudates, and includes
material falling within the definition of "botanical raw material"
in the Guidance for Industry Botanical Drug Products Draft
Guidance, August 2000, US Department of Health and Human Services,
Food and Drug Administration Center for Drug Evaluation and
Research.
[0167] The method of the invention may be used to extract
cannabinoids from a specified and defined plant material known to
contain such cannabinoids. Most typically, but not necessarily, the
"plant material" will be "plant material" or "botanical raw
material" derived from one or more Cannabis plants. Most typically,
but not necessarily, the one or more Cannabis plants will be a
specified and defined Cannabis plant bred to produce a high yield
of CBD.
[0168] The term "Cannabis plant(s)" encompasses wild type Cannabis
sativa and also variants thereof, including Cannabis chemovars
which naturally contain different amounts of the individual
cannabinoids, Cannabis sativa plants which are the result of
genetic crosses, self-crosses or hybrids thereof. The term
"Cannabis plant material" is to be interpreted accordingly as
encompassing plant material derived from one or more Cannabis
plants. For the avoidance of doubt, it is hereby stated that
"Cannabis plant material" includes dried Cannabis biomass. In some
embodiments, at least a portion of the cannabinoid acids in such
Cannabis plant material are decarboxylated.
Cannabis Plants
[0169] The present invention utilizes Cannabis plants and varieties
bred to have specified, predetermined cannabinoid profiles and
content. In some embodiments, a cannabinoid may be CBD, THC, CBDA,
CBDV, CBD-C1, or CBD-C4. In some embodiments, Cannabis plants have
specified, predetermined terpene profiles and content. In some
embodiments, Cannabis plants have specified, predetermined
sesquiterpene profiles and content. In some embodiments, a Cannabis
plant is a Cannabis sativa, Cannabis indica, or Cannabis ruderalis
plant.
Cannabis Cultivation
[0170] In some embodiments, Cannabis plants are propagated from
cuttings taken from a mother plant. In some embodiments, a mother
plant originates from a single seed source. In some embodiments, a
crop is produced through asexual propagation. In some embodiments,
all of the plants in a crop are all female. In some embodiments,
propagation using cuttings controls genotype consistency.
[0171] In some embodiments, the growing cycle is about 12 weeks. In
some embodiments, through controlled growing conditions Cannabis
plants take about 12 weeks to reach maturity. In some embodiments,
Cannabis plants are irrigated throughout their growing cycle with
potable quality water. In some embodiments, no synthetic herbicides
or pesticides are used in the cultivation of Cannabis plants. In
some embodiments, stringent hygiene conditions may be utilized to
reduce ingress of pests and diseases, particularly in the absence
of herbicides or pesticides. In some embodiments, control of
growing conditions to reduce or eliminate environmental stresses is
utilized to optimize plant material yield, cannabinoid content,
and/or control disease. In some embodiments, environmental stresses
may include drought, insufficient light, improper timing of light
cycle, and unfavourable temperatures. In addition, regular
inspection of the plants during the growing cycle allows for the
detection of any rogue plants and pests. Rogue male plants may
arise, though weeds should be absent due to the controlled growing
conditions and media. Frequent inspections and biological control
methods are used to manage any pests and diseases that may
occur.
[0172] In some embodiments, through strict control of growing
conditions the Cannabis plants reach maturity in approximately 12
weeks. In some embodiments, in the last weeks of growth, dense
resinous flowers develop. In some embodiments, by the end of
approximately week 11 the cannabinoid biosynthesis has slowed
markedly, and the plants are ready for harvest.
Cannabis Harvest and Processing
[0173] In some embodiments, the entire plant is cut and dried in a
temperature and/or humidity controlled environment. In some
embodiments, the temperature is about 21.degree. C. In some
embodiments, the humidity is about 38-45% RH.
[0174] THC and CBD are the principle bioactive constituents in the
BDS. However, these constituents are present as the carboxylic
acids THCA and CBDA in the BRM. The acid forms slowly decarboxylate
over time during drying. The leaves and flowers are stripped from
the larger stems to provide the Botanical Raw Material (BRM). Under
conditions of storage the loss on drying reaches equilibrium of
approximately 10%. The storage conditions for the dried BRM will be
dependent on the physical status of the BRM. In some embodiments,
BRM is stored protected from light. In some embodiments, BRM is
stored at about 15-25.degree. C. In some embodiments, BRM is stored
at about -20 .degree. C. In some embodiments, BRM is stored at
about 20.degree. C. In some embodiments, BRM is stored at about
38-42% RH.
[0175] Summary of exemplary production of a BRM: [0176] Harvest
plants [0177] Drying (in absence of light) [0178] Production of
Botanical Raw Material (BRM) which comprises cannabinoid acids
[0179] Milling to less than 2000 .mu.m to reduce particle size
[0180] Decarboxylation of cannabinoid acids to their neutral form
(e.g. CBDA to CBD)
[0181] An exemplary BRM specification derived from a high CBD
variety is illustrated in Table A below:
TABLE-US-00001 TABLE A Exemplary BRM specification Test Method
Specification Identification: A Visual Complies B TLC Corresponds
to standard C HPLC/UV (for CBD & CBDA) Positive for CBDA Assay:
In-house NLT 90% of assayed CBDA + CBD (HPLC/UV) cannabinoids by
peak area Loss on Drying Ph. Eur. NMT 15% Aflatoxin UKAS method NMT
4 ppb Microbial: Ph. Eur. NMT10.sup.7 cfu/g TVC NMT10.sup.5 cfu/g
Fungi NMT10.sup.2 cfu/g E. coli Foreign Matter: Ph. Eur. NMT 2%
Residual Herbicides Ph. Eur. Complies and Pesticides
Characterization of CBD Preparations
Identification by Visual:
[0182] Macroscopic characteristics allow the features of the
Cannabis plant to be distinguished from potential adulterants and
substitutes. It is a visual identification against a photographic
standard.
Identification by TLC:
[0183] TLC uses both retention value of the substance (Rf) and
characteristic spot colour to effectively identify the variety of
Cannabis. Laboratory samples are prepared for TLC analysis by
extracting the dried herb. An aliquot is spotted onto a TLC plate,
alongside reference samples for THC and CBD. Following exposure to
Fast Blue B reagent, THC and THCA present as pink spots, while CBD
and CBDA are orange in colour. Neutrals can be distinguished from
the acids by comparison of the Rf value to that obtained for the
standards. Identity is confirmed by comparison of Rf and colour of
the sample spot, to that obtained for the appropriate standard.
Identification by HPLC:
[0184] HPLC uses retention time comparison of cannabinoids to
effectively identify the variety of Cannabis. The reversed phase
HPLC method is specific for CBD and CBDA, and therefore may be used
as an identity test. Samples of biomass are extracted and
centrifuged. Detection of all analytes is accomplished at 220 nm
with additional confirmation of acidic analytes at 310 nm.
Assay (CBD+CBDA):
[0185] This assay may be used to monitor the CBD and CBDA content
in the plant. CBD and CBDA assay are determined using an HPLC
method. The efficiency of the decarboxylation process may be
determined by dividing the % content in terms of w/w of CBD by the
total CBD+CBDA content.
Foreign Matter:
[0186] Foreign Matter is evaluated using the Ph. Eur. test method.
Flowers, leaves and side stems are spread out in a thin layer on a
clean laboratory surface. Foreign Matter is separated by hand as
completely as possible and is weighed. Results are expressed as %
w/w of Foreign Matter in the herbal biomass sample. Foreign Matter
may comprise no more than 2% of the biomass.
Decarboxylation
[0187] THC and CBD are the principle bioactive constituents in
Cannabis. However, these constituents are present as their
respective carboxylic acids in Cannabis plants. In order to extract
THC or CBD from Cannabis plant material, it is necessary to convert
the storage precursor compounds of THCA and CBDA into their more
readily extractable and pharmacologically active forms. THC and CBD
acids slowly decarboxylate naturally over time. The traditional way
to increase rate of decarboxylation is by the application of heat.
However, THCA is converted not only to THC, but also to another
cannabinoid, cannabinol (CBN).
[0188] The decarboxylation procedure is generally carried out
within the preparation of the starting material or botanical raw
material (BRM), prior to the initiation of the extraction
process.
Overview of Exemplary Extraction Process:
[0189] The BDS may be extracted from decarboxylated BRM using
liquid carbon dioxide methodology. This involves continuously
passing liquefied carbon dioxide through the chopped biomass, which
is contained in a high-pressure vessel. The crude extract is
dissolved in ethanol, cooled to a low temperature then filtered to
remove precipitated constituents such as waxes. Removing ethanol
and water in vacuo produces BDS containing either high
concentrations of CBD or THC, depending on the biomass used.
[0190] Additional methods regarding the purification and
characterization of CBD preparations are disclosed and described in
EP 2 311 475, the content of which is hereby incorporated by
reference in its entirety.
Compositions and Formulations
[0191] CBD preparations may be formulated based on the mode of
intended administration. For example, in some embodiments,
administration may be ocular, oral, parenteral, topical, etc. In
some embodiments, a CBD preparation may be formulated with one or
more excipients to increase stability, increase shelf-life, or
increase efficacy. In some embodiments, a CBD preparation is
formulated for oral administration. In some embodiments a CBD
preparation comprises sesame oil. In some embodiments a CBD
preparation comprises ethanol. In some, embodiments, the ethanol is
ethanol anhydrous. In some embodiments, a CBD preparation comprises
a flavoring. In some embodiments, the flavoring may be a sweetener.
In some embodiments, the sweetener may be an artificial sweetener,
e.g., saccharin, acesulfame, aspartame, neotame, or sucralose. In
some embodiments, the flavoring may be an artificial flavor. In
some embodiments, the artificial flavor may be, e.g., vanilla,
lemon, orange, lime, grapefruit, yuzu, sudachi, apple, pear, peach,
grape, blueberry, strawberry, raspberry, cherry, plum, prune,
raisin, cola, guarana, neroli, pineapple, apricot, banana, melon,
apricot, ume, cherry, raspberry, blackberry, tropical fruit, mango,
mangosteen, pomegranate, papaya, combinations thereof, or the
like.
[0192] Cannabinoid preparations disclosed herein may be formulated
for administration according to methods known in the art.
Uses
Diseases, Disorders, and Conditions
[0193] CBD preparations disclosed herein are useful in providing
analgesia, neuroprotection, reduce inflammation, help alleviate
nausea and emesis, as well as treat epilepsy, anxiety disorders,
and glaucoma. Furthermore, CBD preparations disclosed herein are
useful in providing treatment or amelioration of symptoms in
patients suffering from neurological dysfunction or the
co-morbidities associated with such disorders. In some embodiments,
CBD preparations disclosed herein are more effective in treating
these disorders than prior CBD compositions. In some embodiments, a
CBD preparation of the present invention may be administered in a
lower dose of CBD than a synthetic CBD preparation comprising the
same or similar concentrations of CBD.
[0194] Pain is a common clinical problem confronting all
clinicians. Millions of people in the United States suffer from
severe pain that, according to numerous recent reports, is
chronically under-treated or inappropriately managed. Similarly,
millions of people also suffer from severe nausea and/or frequent
emesis. Moreover, all too frequently, many patients suffering from
chronic, under-treated or irretraceable pain also suffer from lack
of appetite, nausea and/or frequent emesis. These patients present
a greater clinical challenge as they are unable to receive
effective doses of oral pain medications, thereby leaving their
pain unalleviated. Moreover, CBD preparations disclosed herein can
reduce a patient's nausea and vomiting, independent of any pain
relief achieved. Thus, the disclosed CBD preparations are
particularly useful in patients experiencing nausea and vomiting
secondary to un- or under-treated pain. In some embodiments, CBD
preparations disclosed herein are more effective in alleviating
pain than prior CBD compositions.
[0195] A notable percentage of the United States population satisfy
the diagnostic criteria for alcohol use disorders ("AUDs"). The
consumption of excessive amounts of alcohol results in a complex
array of pharmacological effects that directly impact the ability
to treat the condition. These effects directly impact the brain and
include progressive neurodegeneration, impaired executive function
and dependence leading to withdrawal-induced negative effects. CBD
preparations disclosed herein have neuroprotective, anxiolytic and
anti-convulsant effects, which may be effective in preventing
additional brain damage in persons with AUDs, while simultaneously
decreasing the frequency of relapses. In some embodiments, CBD
preparations disclosed herein are more effective in treating these
disorders than prior CBD compositions.
[0196] Chronic abusers of Cannabis can develop dependence and
experience withdrawal symptoms when they attempt to discontinue use
of the drug. Collectively Cannabis dependence and withdrawal are
referred to herein as Cannabis use disorders. CBD preparations
disclosed herein are useful in treating Cannabis use disorders. In
some embodiments, CBD preparations disclosed herein are more
effective in treating these disorders than prior CBD
compositions.
[0197] Dystonia is a neurological movement disorder, with many
known causes, and characterized by involuntary, continual muscular
contractions causing twisting and repetitive movements or abnormal
postures. In some embodiments, CBD preparations disclosed herein
are useful to reduce the muscular contractions characteristic of
this disorder. CBD preparations disclosed herein are more effective
in treating these disorders than prior CBD compositions.
[0198] The etiological pathology of many diseases relates to the
inflammatory processes that are regulated by an individual's immune
system. Inflammation may result from (1) an otherwise appropriate
immunoresponse to an outside trauma, such as brain swelling
secondary to a closed head injury; (2) an overactive
immunoresponse, such as an allergic reaction or dermatitis; or (3)
an inappropriate auto-immunoresponse, such as certain forms of
multiple sclerosis, inflammatory bowel disorders and arthritis.
Regardless of the underlying cause of the inflammation, it is
therapeutically desirable under these circumstances to regulate the
immune system and lessen the inflammatory response. CBD
preparations disclosed herein can regulate various steps in the
immune response and could show some therapeutic benefit in the
treatment of certain inflammatory diseases such as psoriatic
arthritis. In some embodiments, CBD preparations disclosed herein
are more effective in treating these disorders than prior CBD
compositions.
[0199] Rheumatoid arthritis affects approximately 0.5-1% of the
United States population, and autoimmune diseases in general affect
more than 20 million Americans. The pain associated with rheumatoid
arthritis can often be disabling. Cannabinoids have been found to
be useful as an adjunct treatment for rheumatoid arthritis and
joint pain secondary to other autoimmune diseases, such as
inflammatory bowel disease, multiple sclerosis and systemic lupus
erythematosus. In some embodiments, CBD preparations disclosed
herein are more effective in treating these disorders than prior
CBD compositions.
[0200] In addition to the above-discussed therapeutics benefits,
cannabinoids, such as CBD and CBD prodrugs, present a variety of
pharmacological benefits, including, but not limited to,
anti-inflammatory, anti-convulsant, anti-psychotic, antioxidant,
neuroprotective, anti-cancer and immunomodulatory effects. CBD
preparations disclosed herein are more effective in treating these
disorders than prior CBD compositions.
[0201] The present invention provides CBD preparations and
compositions and uses for treating and/or preventing any of a
variety of diseases, disorders, and/or conditions, including, but
not limited to those disclosed herein. In some embodiments, the
present invention provides CBD preparations and compositions and
uses for treating and/or preventing diseases, disorders, or
conditions associated with neurological dysfunction or
neuro-differentiation. In some embodiments, diseases, disorders, or
conditions associated with neurological dysfunction or
neuro-differentiation are those in which neural development is
defective. Such diseases, disorders or conditions are often related
to the neural plasticity of the brain and can include but is not
limited to seizure disorders such as epilepsy. Said seizure
disorders are often associated with co-morbidities such as
cognitive and psychiatric impairment which may be due to the
seizures themselves or the medications used to treat the seizures.
Co-morbid conditions that are known to occur in seizure disorders
include but are not limited to musculoskeletal system disorders;
gastrointestinal and digestive disorders; respiratory system
disorders; chronic pain disorders; cerebrovascular accidents;
migraine; neoplasia; arthritis/rheumatism; obesity; diabetes;
infections; fractures; and allergies. Psychiatric conditions such
as depression; anxiety; autism spectrum disorders; interictal
dysphoric disorder; interictal behavior syndrome; and psychosis of
epilepsy. Cognitive conditions such as cognitive dysfunction;
language abilities; socialization; attention-deficit hyperactivity
disorder; learning disability; mental retardation; and Alzheimer's
disease/dementia.
[0202] In some embodiments, the disease or disorder is a seizure
disorder. In some embodiments, the seizure disorder is epilepsy,
Dravet syndrome, Lennox Gastaut syndrome, febrile infection related
epilepsy syndrome (FIRES), Doose syndrome, Sturge Weber syndrome,
CDKL5 mutation; Aicardi syndrome; bilateral polymicrogyria; Dup15q;
SNAP25; benign rolandic epilepsy; juvenile myoclonic epilepsy;
infantile spasm (West syndrome); and Landau-Kleffner syndrome,
refractory epilepsy, juvenile spasms, West syndrome, infantile
spasms, refractory infantile spasms, tuberous sclerosis complex
(TSC); neurogenetic storage disorder, neuronal ceroid
lipofuscinoses (NCL), Batten disease, brain abnormality, atonic,
idiopathic, absence seizure, partial seizure, simple partial
seizure, or complex partial seizure.
[0203] In some embodiments, the disease or disorder is a
neurodegenerative disease; Alzheimer's disease; Parkinson's
disease; essential tremor; amyotrophic lateral sclerosis (ALS);
Huntington's disease; Friedreich's ataxia; multiple sclerosis;
frontotemporal dementia; prion disease; Lewy body dementia;
progressive supranuclear palsy; vascular dementia; normal pressure
hydrocephalus; traumatic spinal cord injury; HIV dementia; alcohol
induced neurotoxicity; Down's syndrome; movement disorders of the
central and/or peripheral nervous system; motor neurone diseases
(MND); spinal muscular atrophy; or any other related neurological
or psychiatric neurodegenerative disease; brain damage; brain
injury; brain dysfunction; dysgraphia; dysarthria; apraxia;
agnosia; amnesia; dizziness; vertigo; coma; stroke; spinal cord
damage; spinal cord injury; spinal cord disorders; central
neuropathy; peripheral neuropathy; cranial nerve disorder;
trigeminal neuralgia; tumors of the nervous system; infections of
the brain or spinal cord; encephalitis; meningitis; prion disease;
complex regional pain syndrome; an autonomic nervous system
disorder; autonomic neuropathy; dysautonomia; postural orthostatic
tachycardia syndrome (POTS); neurocardiogenic syncope (NCS);
multiple system atrophy (MSA); hereditary sensory and autonomic
neuropathy (HSAN); Holmes-Adie syndrome (HAS); a sleep disorder;
narcolepsy; pain; migraine; cluster headache; tension headache;
back pain; lower back pain; neck pain; neuropathic pain; cancer
pain; allodynia; arthritic pain; inflammatory pain; a
neuropsychiatric disorder; attention deficit hyperactivity
disorder; autism; Tourette's Syndrome; obsessive compulsive
disorder; an autism spectrum disorder; Rett syndrome; Fragile X
syndrome; Angelman syndrome; hyperkinetic disorder; Tourette
syndrome; dystonia; a cancer; brain cancer; glioma; breast cancer;
liver cancer; lung cancer; pancreatic cancer; melanoma; ovarian
cancer; gastric cancer; renal cancer; bladder cancer; addiction;
nicotine addiction; smoking; alcohol addiction; drug addiction;
Cannabis use disorder; a mental disorder; post-traumatic stress
disorder; anxiety; early psychosis; schizophrenia; a cognitive
disorder; stroke; cardiac ischemia; coronary artery disease;
thromboembolism; myocardial infarction; ischemic related disease; a
gastrointestinal disorder; inflammatory bowel disease; Crohn's
disease; ulcerative colitis; nausea; vomiting; emesis; motion
sickness; chemotherapy induced nausea; chemotherapy induced nausea
vomiting; inflammation; arthritis; rheumatoid arthritis;
osteoarthritis; diabetes; high blood pressure; poor insulin
control; appetite suppression; anorexia; neonatal hypoxic-ischemic
encephalopathy (NHIE); a degenerative skeletal muscle disease; or
Duchenne muscular dystrophy (DMD).
Dosing and Administration
[0204] The exact regimen for administration of the compounds
described herein may depend on the needs of the individual subject
being treated, the type of treatment administered, and/or the
judgment of the attending medical specialist. As used herein, the
terms "subject" and "patient" includes both humans and animals. In
some embodiments, the subject or patient is a human adult, human
adolescent, human child, or human infant. As those skilled in the
art will appreciate, the dosage administered will depend upon the
condition being treated, the age, health and weight of the
recipient, the type of concurrent treatment, if any, and the
frequency of treatment.
[0205] In some embodiments, a CBD preparation or a pharmaceutical
composition comprising CBD may be administered in a therapeutically
effective amount. A therapeutically effective amount may be
administered according to a dosing regimen comprising one or more
unit doses. Generally, a therapeutically effective amount is
sufficient to achieve a benefit to the subject (e.g., prophylaxis,
treating, modulating, curing, preventing and/or ameliorating a
disease or disorder).
[0206] A therapeutically effective amount (and/or unit dose) of a
CBD preparation or a pharmaceutical composition comprising the same
for any particular patient may depend upon a variety of factors
including the disease or disorder being treated; disease or
disorder severity; the activity of the specific CBD preparation or
a pharmaceutical composition comprising the same employed; the
specific CBD preparation or a pharmaceutical composition comprising
the same employed; the age; body weight; fitness; comorbid
conditions (e.g., other than the diseases or disorder(s) being
treated) general health; sex; and diet of the patient; personal
history; genetic characteristic; lifestyle parameter; severity of
cardiac defect and/or level of risk of cardiac defect; the time of
administration; route of administration; concomitant treatments or
medications; and/or rate of excretion or metabolism of the specific
CBD preparation or a pharmaceutical composition comprising the same
employed; the duration of the treatment; combinations thereof; as
well as other factors well known in the medical arts. In view of
the present disclosure, one of ordinary skill in the art will be
readily able to determine appropriate dosages depending on these
and other related factors. In addition, both objective and
subjective assays may optionally be employed to identify optimal
dosage ranges. In some particular embodiments, appropriate doses or
amounts to be administered may be extrapolated in view of the
instant disclosure from dose-response curves derived from in vitro
or animal model test systems.
[0207] The present invention contemplates dosing regiments
comprising single as well as multiple administrations of a CBD
preparation or a pharmaceutical composition comprising the same
described herein. A CBD preparation or a pharmaceutical composition
comprising the same can be administered at regular intervals,
depending on the nature, severity and extent of the subject's
condition. In some embodiments, a CBD preparation or a
pharmaceutical composition comprising the same may be administered
periodically at regular intervals (e.g., once every year, once
every six months, once every five months, once every three months,
bimonthly (once every two months), monthly (once every month),
biweekly (once every two weeks), weekly, daily, multiple times each
day, or continuously).
[0208] A therapeutically effective amount may be administered
according to a dosing regimen that may comprise multiple unit
doses. For any particular CBD preparation or a pharmaceutical
composition comprising the same, a therapeutically effective amount
(and/or an appropriate unit dose within an effective dosing
regimen) may vary, for example, depending on route of
administration, on combination with other pharmaceutical
agents.
[0209] In some embodiments, a CBD preparation or a pharmaceutical
composition comprising the same described herein may be
administered as a single dose. In some embodiments, a CBD
preparation or a pharmaceutical composition comprising the same
described herein may be administered at regular intervals.
Administration at an "interval," as used herein, indicates that the
therapeutically effective amount is administered periodically (as
distinguished from a one-time dose). The interval can be determined
by standard clinical techniques. In some embodiments, a CBD
preparation or a pharmaceutical composition comprising the same
described herein may be administered bimonthly, monthly, twice
monthly, triweekly, biweekly, weekly, twice weekly, thrice weekly,
daily, twice daily, every six hours, every four hours, every two
hours, or hourly. The administration interval for a given
individual need not be a fixed interval, but may be varied over
time, depending on the needs of the individual.
[0210] In some embodiments, a CBD preparation or a pharmaceutical
composition comprising the same described herein is administered at
regular intervals indefinitely. In some embodiments, a CBD
preparation or a pharmaceutical composition comprising the same
described herein is administered at regular intervals for a defined
period.
[0211] It is to be further understood that for any particular
subject, specific dosage regimens should be adjusted over time
according to the individual need and the professional judgment of
the person administering or supervising the administration of the
enzyme replacement therapy and that dosage ranges set forth herein
are exemplary only and are not intended to limit the scope or
practice of the claimed invention.
[0212] In some embodiments, a CBD preparation or a pharmaceutical
composition comprising the same is administered as one or more
doses to provide about 0.1 mg/kg/day of CBD. In some embodiments, a
CBD preparation or a pharmaceutical composition comprising the same
is administered as one or more doses to provide about 0.5 mg/kg/day
of CBD. In some embodiments, a CBD preparation or a pharmaceutical
composition comprising the same is administered as one or more
doses to provide about 1 mg/kg/day of CBD. In some embodiments, a
CBD preparation or a pharmaceutical composition comprising the same
is administered as one or more doses to provide about 5 mg/kg/day
of CBD, e.g., for a 15 kg patient, 75 mg of CBD per day would be
provided. In some embodiments, a CBD preparation or a
pharmaceutical composition comprising the same is administered as
one or more doses to provide about 10 mg/kg/day of CBD. In some
embodiments, a CBD preparation or a pharmaceutical composition
comprising the same is administered as one or more doses to provide
about 20 mg/kg/day of CBD. In some embodiments, a CBD preparation
or a pharmaceutical composition comprising the same is administered
as one or more doses to provide about 25 mg/kg/day of CBD. In some
embodiments, a CBD preparation or a pharmaceutical composition
comprising the same is administered as one or more doses to provide
about 50 mg/kg/day of CBD. In some embodiments, a CBD preparation
or a pharmaceutical composition comprising the same is administered
as one or more doses to provide about 100 mg/kg/day of CBD.
[0213] In some embodiments, a CBD preparation or a pharmaceutical
composition comprising the same is administered in a dose of about
1 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40
mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90
mg, about 100 mg, about 150 mg, about 200 mg, about 300 mg, about
400 mg, about 500 mg, about 1000 mg, about 1500 mg, or about 2000
mg of CBD.
[0214] In some embodiments a CBD preparation or a pharmaceutical
composition comprising the same may be administered as an adjunct
to conventional therapy for a disease or disorder.
Kits
[0215] Some embodiments provide for a kit comprising a CBD
preparation or a pharmaceutical composition comprising the same and
instructions for use. In some embodiments a kit further comprises a
device (e.g., spray, syringe, vaporizer, inhaler, patch, etc.) for
administration of said CBD preparation or pharmaceutical
composition comprising the same.
EXAMPLES
Data Demonstrating Physicochemical Properties of Botanically
Derived Purified CBD
Example 1: Exemplary Process for Production of a Botanically
Derived Purified CBD Preparation
Overview of the Process
[0216] The following describes the production of the botanically
derived purified CBD (>98% w/w) which has a known and constant
composition which was used in the Examples below.
[0217] Plant material harvested from the Cannabis sativa L. plant
was subjected to liquid carbon dioxide extraction, to produce a
botanical extract containing CBD in addition to other cannabinoids
and non-cannabinoid components. The extract was then further
purified by a solvent crystallization method to yield botanically
derived purified CBD. The crystallization process specifically
removed other cannabinoids and plant components to yield greater
than 98% (w/w) CBD.
[0218] Both the botanical starting material and the botanical
extract may be controlled by specifications. An exemplary botanical
starting material specification for decarboxylated Cannabis plant
material is described in Table 1.1 below. In some embodiments, the
isomeric content for each cannabinoid may also be specified.
TABLE-US-00002 TABLE 1.1 Exemplary Botanical Starting Material
Specification Decarboxylated % w/w BRM CBD THC CBD-C1 CBDV CBD-C4
MEAN 11.96 0.46 0.03 0.14 0.05 MIN 6.90 0.27 0.01 0.08 0.03 MAX
16.93 0.65 0.04 0.20 0.07 SD 2.30 % RSD 19.20 MEAN -3SD 5.07 MEAN
+3SD 18.85 MEDIAN 12.00 COUNT 693 approximate ratio CBD to 26 475
85 230 other cannabinoid
[0219] An exemplary CBD preparation of botanically derived purified
CBD is described in Table 1.2 below. In some embodiments, the
isomeric content for each cannabinoid may also be specified.
TABLE-US-00003 TABLE 1.2 Specification of an exemplary botanically
derived purified CBD preparation Test Test Method Limits Appearance
Visual Off-white/pale yellow crystals Identification HPLC-UV
Retention time of major peak A corresponds to certified CBD
Reference Standard Identification GC-FID/MS Retention time and mass
spectrum B of major peak corresponds to certified CBD Reference
Standard Identification FT-IR Conforms to reference spectrum for C
certified CBD Reference Standard Identification Melting
65-67.degree. C. D Point Identification Specific Conforms with
certified CBD E Optical Reference Standard; -110.degree. to
-140.degree. Rotation (in 95% ethanol) Total Purity Calculation
.gtoreq.98.0% Chromatographic HPLC-UV .gtoreq.98.0% Purity 1
Chromatographic GC-FID/MS .gtoreq.98.0% Purity 2 CBDA HPLC-UV NMT
0.15% w/w CBDV NMT 1.0% w/w .DELTA..sup.9 THC NMT 0.1% w/w CBD-C4
NMT 0.5% w/w Residual Solvents: GC NMT 0.5% w/w Alkane NMT 0.5% w/w
Ethanol Residual Water Karl Fischer NMT 1.0% w/w i.NMT--Not more
than
[0220] The purity of the botanically derived purified CBD
preparation was greater than or equal to 98%. The botanically
derived purified CBD includes THC and other cannabinoids, e.g.,
CBDA, CBDV, CBD-C1, and CBD-C4.
[0221] Distinct chemotypes of the Cannabis sativa L. plant have
been produced to maximize the output of the specific chemical
constituents, the cannabinoids. Certain chemovars produce
predominantly CBD. Only the (-)-trans isomer of CBD is believed to
occur naturally. During purification, the stereochemistry of CBD is
not affected.
Production of CBD Botanical Drug Substance
[0222] An overview of the steps to produce a botanical extract, the
intermediate, are as follows: [0223] a. Growing [0224] b. Direct
drying [0225] c. Decarboxylation [0226] d. Extraction--using liquid
CO.sub.2 [0227] e. Winterization using ethanol [0228] f. Filtration
[0229] g. Evaporation
[0230] High CBD chemovars were grown, harvested, dried, baled and
stored in a dry room until required. The botanical raw material
(BRM) was finely chopped using an Apex mill fitted with a 1 mm
screen. The milled BRM was stored in a freezer prior to
extraction.
[0231] Decarboxylation of CBDA to CBD was carried out using heat.
BRM was decarboxylated at 115.degree. C. for 60 minutes.
[0232] Extraction was performed using liquid CO.sub.2 to produce
botanical drug substance (BDS), which was then crystalized to
produce the test material. The crude CBD BDS was winterized to
refine the extract under standard conditions (2 volumes of ethanol
at -20.degree. C. for approximately 50 hours). The precipitated
waxes were removed by filtration and the solvent was removed to
yield the BDS.
Production of Botanically Derived Purified CBD Preparation
[0233] The manufacturing steps to produce the botanically derived
purified CBD preparation from BDS were as follows: [0234] a.
Crystallization using C.sub.5-C.sub.12 straight chain or branched
alkane [0235] b. Filtration [0236] c. Vacuum drying
[0237] The BDS produced using the methodology above was dispersed
in C.sub.5-C.sub.12 straight chain or branched alkane. The mixture
was manually agitated to break up any lumps and the sealed
container then placed in a freezer for approximately 48 hours. The
crystals were isolated via vacuum filtration, washed with aliquots
of cold C.sub.5-C.sub.12 straight chain or branched alkane, and
dried under a vacuum of <10 mb at a temperature of 60.degree. C.
until dry. The botanically derived purified CBD preparation was
stored in a freezer at -20.degree. C. in a pharmaceutical grade
stainless steel container, with FDA food grade approved silicone
seal and clamps.
Example 2: Quantification of the Stereochemistry of the THC Present
in Botanically Derived Purified CBD Preparations
[0238] As described in Example 1 above, the cannabinoid THC is
present in the botanically derived purified CBD preparation at an
amount of not more than 0.1% (w/w). In certain embodiments the THC
is present at an amount of 0.02 to 0.1% w/w.
[0239] The THC molecule may exist as four distinct chiral forms as
shown in FIG. 2. THC has 2 stereocenters which in turn enable the
existence of 4 stereoisomers: (+)-trans-THC; (-)-trans-THC;
(+)-cis-THC and (-)-cis-THC. However, THC is known to be produced
in nature as the (-)-trans-THC isoform (Hollister, 1970).
[0240] The following examples describe studies undertaken to
elucidate the stereoisomers which are produced by the high-CBD
plant used to prepare the exemplary botanically derived purified
CBD preparation which is described in Example 1.
Detection and Quantification of Trace Levels of THC in Botanically
Derived Purified CBD
[0241] Ultra Performance Liquid Chromatography (UPLC) was used to
identify the principal cannabinoid and trace levels of other
cannabinoids. The presence of the cannabinoids was confirmed by
chromatographic comparison of the sample with the standard.
[0242] A core shell silica column with adjusted pH was used in a
UPLC System fitted with a Photo Diode Array or Tuneable UV detector
and QDa mass detector.
[0243] FIG. 3 demonstrates an exemplary chromatogram produced using
the UPLC method to detect trace quantities of cannabinoids in the
botanically derived purified CBD preparation of the present
invention.
[0244] As can be seen in FIG. 3 there are peaks which correspond to
the following cannabinoids: CBD-C1 at a relative retention time
(RTT) of 1.937 minutes; CBDV at an RTT of 3.481 minutes; CBD-C4 at
an RTT of 4.610 minutes; CBD at an RTT of 5.920 minutes;
(.+-.)-cis-THC at an RTT of 6.781 minutes and (.+-.)-trans-THC at
an RTT of 6.917 minutes.
[0245] The concentration of the compounds present in the sample can
be determined as % w/w using the following calculation:
Concentration ( % w / w ) = Peak area .times. concentration in
standard ( mg / ml ) .times. dilution factor Area im standard
.times. sample weight ( mg ) ##EQU00001##
[0246] The presence of cis-THC in the exemplary botanically derived
purified CBD preparation was surprising as it is known that THC
produced by the THC plant is in the form of (-)-trans-THC. As such
the structure elucidation of the cis-THC was undertaken.
Structure Elucidation of Cis-THC
[0247] The presence, isolation and identification of cis-THC was
determined from the botanical raw material produced from a high-CBD
plant and in consequence the resulting botanically derived purified
CBD preparation as described in Example 1 was required.
[0248] The cis-THC was successfully isolated and purified from
cannabidiol (CBD) botanical drug substance (BDS) using flash
chromatography and preparative-LCMS. The isolated material
underwent extensive testing, including chromatographic and spectral
techniques, alongside a synthetic cis-THC standard to identify the
compound. It was also compared to its configurational isomer;
trans-THC.
[0249] The identity of cis-THC was confirmed by four spectral
techniques and three chromatographic techniques. The spectra of the
botanically isolated material compared to the synthetic sample
which confirmed the structure to be cis-THC.
[0250] The identity of botanically derived cis-THC was confirmed by
the following techniques: [0251] Determination of physical data and
interpretation of molecular spectroscopy results; including:
1H-NMR, COSY-NMR, mass spectrometry and Quadrupole Time of Flight.
[0252] Comparing physical and spectral data of synthetically
produced cis-THC to that of the botanically isolated material. This
was further confirmed by comparison to the previously proposed
structure of cis-THC from the literature.
[0253] FIG. 4 details a trace produced using the mass spectrometer
which confirms the presence of cis-THC which had been isolated from
high-CBD plant material.
Example 3: Quantification of the Ratio of Trans-THC to Cis-THC
Present in Botanically Derived Purified CBD Preparations
[0254] As is described in Example 1, the production of a
botanically derived purified CBD preparation is a complex process
containing several steps resulting in different types of
intermediate materials. Firstly, the high-CBD plants are harvested,
dried and baled to produce Botanical Raw Material, (BRM). The baled
material is pelleted, extracted using liquid CO.sub.2, refined
during the winterization process and purified during the
crystallization process. The CBD BRM starting material contains a
number of different cannabinoid impurities present alongside the
principal compound CBD.
[0255] The present example sought to determine the ratio of
trans-THC to cis-THC throughout the production process of the
botanically derived purified CBD preparation.
[0256] The retention time of trans-THC and cis-THC is different
using reverse phase chromatography as shown in FIG. 5. The polarity
of these two compounds is slightly different which results in close
eluting peaks in an isocratic method with baseline resolution.
[0257] The ratio of trans-THC to cis-THC was determined in samples
obtained from material in different phases of the process used to
produce the botanically derived purified CBD preparation described
in Example 1.
[0258] The ratio of trans-THC to cis-THC changed throughout the
process from the decarboxylated CBD to the botanically derived
purified CBD preparation as is described in Table 3.1 below.
TABLE-US-00004 TABLE 3.1 Ratio of trans-THC to cis-THC in CBD
material during processing stages Median ratio 1st Quartile 3rd
Quartile (trans- (trans- (trans- THC:cis- THC:cis- THC:cis- CBD
Sample THC) THC) THC) Decarboxylated CBD 3.6:1 3.1:1 3.8:1 CO.sub.2
extracted CBD 3.3:1 3.0:1 3.6:1 Winterized CBD 3.2:1 3.0:1 3.6:1
Botanically derived 0.8:1 0.7:1 1.0:1 purified CBD
[0259] As can be seen in Table 3.1 above the ratio of trans-THC to
cis-THC changes throughout the processing of the botanically
derived purified CBD preparation. The botanical raw material once
decarboxylated has a median ratio of 3.6:1 (trans-THC:cis-THC),
this ratio becomes smaller as the material is processed with the
material extracted with liquid carbon dioxide having a median ratio
of 3.3:1 (trans-THC:cis-THC) and the material that had been refined
by the winterization process having a median ratio of 3.2:1
(trans-THC:cis-THC). Finally, the further purification step of
crystallization of the CBD material using a C.sub.5-C.sub.12
straight chain or branched alkane produces a median ratio of 0.8:1
(trans-THC:cis-THC).
[0260] The ratio of trans-THC to cis-THC at the different
processing stages were also plotted using a Spearman rank
correlation curve, these are detailed in FIG. 6. The fit of the
line demonstrates a change in ratio during the processing and has a
very high confidence level. Such a correlation demonstrates control
over the processing stages of the preparation of the botanically
derived purified CBD where retention of the cis-THC isomer is
favored over the trans-THC.
[0261] As can be seen the botanically derived purified CBD
preparation comprises both trans-THC and cis-THC. The ratio of the
two isomers in the highly purified final preparation is 0.8:1
(trans-THC:cis-THC).
Example 4: Quantification of the Ratio of Trans-THC to Cis-THC
Present in Crude CBD Preparations
[0262] In order to determine whether both trans-THC and cis-THC
existed in crude CBD preparations and also if both were present
what the ratio of these were, a further study was undertaken using
CBD oil purchased from CBD-oil vendors.
[0263] Eight different crude CBD oil preparations were tested and
all were found to comprise a mixture of trans-THC and cis-THC.
Table 4.1 below details the ratios that were found in these
oils.
TABLE-US-00005 TABLE 4.1 Ratio of trans-THC to cis-THC in crude CBD
material Mean ratio Sample number (trans-THC:cis-THC) 1 4.4:1 2
2.7:1 3 2.5:1 4 3.2:1 5 2.7:1 6 2.3:1 7 3.6:1 8 2.7:1
[0264] As Table 4.1 demonstrates the ratio of trans-THC to cis-THC
varies within the different crude CBD oil preparations obtained
from 2.3:1 to 4.4:1 (trans-THC:cis-THC). These ratios are far
removed from the botanically derived purified CBD material defined
in the present invention.
Example 5: Quantification of the Stereoisomeric Form of Cis-THC
Present in Botanically Derived Purified CBD Preparation
[0265] Example 3 demonstrates that botanically derived purified CBD
preparations comprise both trans-THC and cis-THC at a median ratio
of 0.8:1 (trans-THC and cis-THC). As described in Example 2 and
FIG. 2, the THC molecule has 2 stereocenters which enable the
existence of four stereoisomers: (+)-trans-THC; (-)-trans-THC;
(+)-cis-THC and (-)-cis-THC. The present example sought to
determine whether the cis-THC was present as the (+)-cis-THC
isomer, the (-)-cis-THC isomer or a mixture of the two.
[0266] A chiral method employing the principals of normal phase
chromatography was developed which could successfully separate
(+)-cis-THC and (-)-cis-THC.
[0267] An amylose carbamate chiral column was used with a mobile
phase employing isocratic heptane with an ethanol modifier.
[0268] A cis-THC sample (98.2% pure by HPLC) was isolated from
botanically derived purified CBD and run on the chiral method, for
the chirality to be assessed. The resulting trace is shown in FIG.
7. As can be seen in Trace C there are peaks present for both
(-)-cis-THC and (+)-cis-THC indicating that the cis-THC present in
botanically derived purified CBD preparations is present as a
mixture of both isomers.
[0269] The approximate ratio of the (-)-cis-THC to (+)-cis-THC
present was calculated to be 9:1, as determined by area under the
curve. It is likely that various ranges of ratios of (-)-cis-THC to
(+)-cis-THC will exist depending on the source and processing of
the CBD material.
Example 6: Quantification of Composition of Synthetically Produced
CBD
[0270] Two different samples of synthetically produced CBD were
obtained and run on an HPLC to compare the composition of CBD of
synthetic origin with that of botanically derived purified CBD.
Both preparations comprised greater than or equal to 98% (w/w)
CBD.
[0271] As can be seen in FIG. 8 there is a difference between the
CBD of synthetic origin and that of botanically derived purified
CBD. The botanically derived purified CBD sample has peaks which
correspond to CBD-C1, CBDV, CBD-C4, trans-THC and cis-THC whereas
the synthetically produced CBD does not comprise these
compounds.
Example 7: Autofluorescence Properties of Botanically Derived
Purified CBD and Synthetic CBD
[0272] The physical properties of the emission and excitation
spectrum of botanically derived purified CBD and synthetic CBD were
determined in this example.
Methods
Test Substances
[0273] Botanically derived purified CBD (BOT) was tested in
duplicates at six concentrations of 100 mM in 100% DMSO or 100%
Ethanol, and at 100 .mu.M, 50 .mu.M, 1 .mu.M, 0.5 .mu.M, and 0.01
.mu.M in PBS+0.1% Ethanol or PBS+0.1% DMSO containing vehicle
solution. The THC concentration in the botanically derived purified
CBD used was 0.03% (w/w) THC.
[0274] Synthetic CBD (SYN) was tested in duplicates at five
concentrations of 100 mM, 50 mM, 1 mM, 0.5 mM, and 0.1 mM, in 100%
Ethanol or 100% DMSO.
Microplate Preparation
[0275] 200 .mu.L of each sample or buffer were added into
microplates in duplicate. Measurement has been performed on the
Ensight multimodal plate reader (Perkin Elmer).
Selection of Excitation Wavelength
[0276] An excitation scan was used to detect the excitation peak
for each test substance. The excitation scan was set from 230 to
380 nm and below the emission wavelength of 400 nm. The excitation
scan was also set from 230 to 420 nm and below the emission
wavelength of 440 nm. In both cases, a step increment of 2 nm was
used for these scans.
[0277] Based on these analyses, the peaks of excitation wavelengths
for the two tentative emission wavelengths were determined and used
to identify the optimal excitation wavelength/s.
Selection of Emission Wavelength
[0278] In order to confirm the emission peak of the test substance,
an emission scan was performed using the excitation peaks
identified after the excitation scan. The excitation scan was set
to start for botanically derived purified CBD at 50 nm above the
tentative excitation values and for synthetic CBD at 20 nm above
the tentative excitation values using a step increment of 2 nm.
Wavelengths up to 800 nm were scanned for emission.
[0279] The wavelength giving the maximum emission for the samples
was defined as the optimal emission wavelength for the chosen
excitation wavelength.
Data Presentation
[0280] Raw data were derived from the software WorkOut Plus
(version 2.5, Perkin Elmer, Waltham, Mass., USA). All data
extracted from the software were checked by two people for complete
verification before data analysis.
[0281] These data were used to generate the data presented herein,
representing in x the wavelength (nm) and in y the fluorescent
intensity using WorkOut Plus software (version 2.5, Perkin
Elmer).
[0282] Data were subsequently to plot these as XY graphs using
GraphPad Prism (version 8.0.2, La Jolla, USA).
Results
Excitation Peak Points for Botanically Derived Purified CBD and
Synthetic CBD in DMSO
[0283] Specific autofluorescence spectra distinct from the DMSO
profile for each compound could only be detected at the highest
concentration of 100 mM.
[0284] Representative spectroscopic data for each compound at the
concentration of 100 mM are shown in FIG. 9.
[0285] Using pure DMSO as vehicle and with emission wavelength set
at 400 or 440 nm, excitation peak points were identified for
botanically derived purified CBD at 328 or 332 nm; with
monodispersed peaks of higher fluorescence reading for botanically
derived purified CBD compared to synthetic CBD.
[0286] Using pure DMSO as vehicle and with emission wavelength set
at 400 or 440 nm, although excitation peak points were identified
for synthetic CBD 282 or 328 nm; these overlap with peaks for DMSO
suggestive of absence of excitation peak for this material in these
conditions.
Emission Peak Points for Botanically Derived Purified CBD and
Synthetic CBD in DMSO
[0287] Specific autofluorescence for each compound could only be
detected at the highest concentration of 100 mM. Data suggested low
specific fluorescence detection similar to that of DMSO, at all
lower concentrations.
[0288] Representative spectroscopic data for each compound at the
concentration of 100 mM are shown in FIG. 10.
[0289] Using pure DMSO as vehicle and with excitation wavelength
set at 326 as identified from this study or 370 nm based on
literature, emission peak points were identified for botanically
derived purified CBD at 398 or 428 nm with a monodispersed peak of
highest fluorescence reading at 326 nm.
[0290] Using pure DMSO as vehicle and with excitation wavelength
set at 328/334/344/284 nm, excitation peak points were identified
for synthetic CBD at 408/498/508/565 nm with a monodispersed peak
of highest fluorescence reading at 565 nm. As this overlaps with
DMSO peak it has not been included in the spectrogram.
Excitation Peak Points for Botanically Derived Purified CBD and
Synthetic CBD in Ethanol
[0291] Specific autofluorescence spectra distinct from the ethanol
profile for each compound could only be detected at the highest
concentration of 100 mM.
[0292] Representative spectroscopic data for each compound at the
concentration of 100 mM are shown in FIG. 11.
[0293] Using pure ethanol as vehicle and with emission wavelength
set at 400 nm, excitation peak points were identified for
botanically derived purified CBD at 328 nm and synthetic CBD at 280
nm.
[0294] Using pure ethanol as vehicle and with emission wavelength
set at 440 nm, excitation peak points were identified for
botanically derived purified CBD at 330 nm and synthetic CBD at 328
nm.
Emission Peak Points for Botanically Derived Purified CBD and
Synthetic CBD in Ethanol
[0295] Specific autofluorescence for each compound could only be
detected at the highest concentration of 100 mM. Data suggested low
specific fluorescence detection similar to that of ethanol, at all
lower concentrations.
[0296] Representative spectroscopic data for each compound at the
concentration of 100 mM are shown in FIG. 12.
[0297] Using pure ethanol as vehicle and with excitation wavelength
set at 326 nm as identified in this study or 370 nm based on the
literature, emission peak points were identified for botanically
derived purified CBD at 404 nm or 560 nm.
[0298] Using pure ethanol as vehicle and with excitation wavelength
set at 326 and 340 nm, excitation peak points were identified for
synthetic CBD between 412 nm for both wavelengths.
Conclusion
[0299] The data presented in this example suggest there is
difference in the excitation and emission wavelengths of
botanically derived purified CBD and synthetic CBD. As such there
is an apparent difference in the biophysical properties of the two
compounds.
Conclusion of Data Demonstrating Physicochemical Properties of
Botanically Derived Purified CBD
[0300] The data presented in Examples 2 to 7 above describe the
physicochemical properties of the botanically derived purified CBD
preparation that was produced from a high-CBD plant.
[0301] This botanically derived purified CBD comprises greater than
or equal to 98% (w/w) CBD and less than or equal to 2% (w/w) of
other cannabinoids. The other cannabinoids present are THC at a
concentration of less than or equal to 0.1% (w/w); CBD-C1 at a
concentration of less than or equal to 0.15% (w/w); CBDV at a
concentration of less than or equal to 0.8% (w/w); and CBD-C4 at a
concentration of less than or equal to 0.4% (w/w).
[0302] These data additionally demonstrate that both trans-THC and
cis-THC are present in the botanically derived purified CBD.
Furthermore, it has been shown that the ratio of the trans-THC to
cis-THC is altered and can be controlled by the processing and
purification process, ranging from 3.3:1 (trans-THC:cis-THC) in its
unrefined decarboxylated state to 0.8:1 (trans-THC:cis-THC) when
highly purified.
[0303] Finally, these data demonstrate that the cis-THC found in
botanically derived purified CBD is present as a mixture of both
the (+)-cis-THC and the (-)-cis-THC isoforms.
[0304] Of note the comparison of the botanically derived purified
CBD differs greatly in composition from that of a synthetic CBD.
However as provided by the teachings of this invention a CBD
preparation could be produced synthetically by producing a
composition with duplicate components.
Data Demonstrating Therapeutic Efficacy of Botanically Derived
Purified CBD
[0305] The following Examples describe data generated in vitro
using cell lines and in vivo using animal studies in particular
disease areas which demonstrate the superior efficacy of the
botanically derived purified CBD preparation over that of synthetic
CBD of the same concentration and purity.
[0306] The purity of the botanically derived purified CBD
preparation used in these experiments was greater than or equal to
98%. The CBD preparation included less than or equal to 2% of other
cannabinoids including THC, CBDV, CBD-C1, and CBD-C4. The THC was
present at a concentration of less than or equal to 0.1% (w/w);
CBD-C1 at a concentration of less than or equal to 0.15% (w/w);
CBDV at a concentration of less than or equal to 0.8% (w/w); and
CBD-C4 at a concentration of less than or equal to 0.4% (w/w).
[0307] The THC is present as both trans-THC and cis-THC in the
botanically derived purified CBD preparation used in the following
Examples. The ratio of the trans-THC to cis-THC was approximately
0.8:1 (trans-THC:cis-THC). The cis-THC was present as a mixture
with both the (+)-cis-THC and the (-)-cis-THC isoforms being
present.
Example 8: In Vitro Study on Neural Plasticity and Growth Cone
Development
[0308] This example demonstrates that certain concentrations of
botanically derived purified CBD are capable of inducing cell
differentiation and neurite outgrowth.
[0309] Studies were conducted in order to identify the effects of
botanically derived purified CBD and synthetic CBD on human iPSC
derived cerebral cortical neural stem/progenitor cells in vitro
(hNSCs).
Methods
Culture of Human iPSC-Derived Cerebral Cortical Neural Stem Cells
(hNSCs)
[0310] Human iPSC-derived cerebral cortical neural stem cells
(hNSCs) (Axol Bioscience Inc., UK) were cultured as monolayers at a
density of 1.0.times.10.sup.4 cells/cm.sup.2 on 100 mm diameter
Petri dishes. The cells were collected from their original
cryovial, suspended in Plating-XF Medium and plated on Petri dishes
coated with SureBondXF 1.times. working solution prepared in D-PBS
(without calcium or magnesium). hNSCs were incubated at 37.degree.
C. and 5% CO.sub.2 and 24 hours after of incubation, the plating
medium was replaced with fresh Neural Expansion-XF Medium
supplemented with the growth factors Recombinant Human EGF and
Recombinant Human FGF2 (final concentration of 20 ng/mL (1.times.)
of FGF2 and EGF). The cultures were re-fed with fresh Neural
Expansion-XF supplemented with EGF and FGF2 every two days. When
the hNSCs reached approximately 80% confluence, they were prepared
for treatment. Pre-warmed Axol Unlock-XF was used to detach the
cells and after 3 minutes of incubation the cells were re-suspended
in Neural Expansion-XF Medium supplemented with EGF and FGF2. The
cells (1.times.10.sup.4 cells/cm.sup.2) were then plated in Petri
dishes coated with SureBondXF 1.times. and after 2 hours the medium
was replaced with fresh Neural Expansion-XF supplemented with EGF
and FGF2.
Cell Treatment with Test Cannabinoids
[0311] hNSCs cultured as described above were plated at a density
of 1.times.10.sup.4 cells/cm.sup.2 on SureBondXF-coated coverslips
in the presence of botanically derived purified CBD; synthetic CBD;
or vehicle diluted in the Neural Differentiation-XF Medium (DM,
without growth factors) for 3-5 days at 37.degree. C. in a 5%
CO.sub.2/95% air atmosphere.
[0312] Cannabinoids were dissolved in 95% (v/v) ethanol and added
to the medium to obtain final concentrations of 0.1 or 1 .mu.M
CBD.
[0313] Ethanol was added to the vehicle-treated cells for the whole
differentiation period of 3-5 days wherein the final concentration
was never higher than 0.05% (v/v).
Immunohistochemistry
[0314] In order to perform immunohistochemistry, the cells were
fixed with 4% (w/v) paraformaldehyde. Nonspecific binding was
blocked with 10% (v/v) fetal bovine serum and 0.1% (v/v) Triton
X-100 and incubated with primary antibodies: mouse anti-Map2ab
(Sigma-Aldrich; 1:200) and rabbit anti-GFAP (DAKO; 1:200) or rabbit
anti-Gap43 (Synaptic System; 1:300 at room temperature for 4 hours.
Secondary antibodies--anti-mouse Alexa594 (Life Technologies;
1:100) and anti-rabbit Alexa488 (Life Technologies; 1:100)--were
used for immunofluorescence. Cells were then counterstained with
DAPI, washed with PBS and cover slipped with Aquatex mounting
medium (Merck, Darmstadt, Germany). The immunofluorescence was
studied with an epifluorescence microscope (Leica AF6000) equipped
with the appropriate filter and then acquired using a digital
camera (Leica, DFC 340) connected to the microscope and image
analysis software (Leica, LAS AF). No immunoreactivity was found in
the samples processed for the control of specificity of each
primary antibody used in the study, which was performed by omitting
the primary antibody before adding the appropriate secondary
antibody.
Data Collection and Statistical Analysis
[0315] Digital Leica LAS AF 2.2.0 software (Live Data Mode system)
was used for counting Map2ab+DAPI+ cells. The images were acquired
with a Leica DMI6000 microscope equipped with appropriate blue
(DAPI) and red (MAP2) filters for counting differentiated cells as
violet (merge of DAPI, blue, and MAP2, red) and hNSCs as blue (DAPI
only). For each experimental condition, cells from 3 different
tubes were each analyzed in triplicate and 3 frames were collected
at 20.times. magnification. Data were analyzed with GraphPad Prism
6 software, version 6.05 (GraphPad, Inc.) and are expressed as
mean.+-.SEM. Statistical differences among groups were determined
by two-way ANOVA followed by post hoc Bonferroni tests for
comparison among means. A level of confidence of P<0.05 was
employed for statistical significance.
Results
Effect of Botanically Derived Purified CBD and Synthetic CBD on
Neurogenesis and Neuronal Differentiation of hNSCs
[0316] Two different botanically derived purified CBD preparations
were tested on cultured human neural stem cells (hNSCs). Batch 1
comprised approximately 0.1% (w/w) THC and Batch 2 comprised 0.02%
THC (w/w). Doses of 0.1 and 1 .mu.M were used with both botanically
derived purified CBD and synthetic CBD.
[0317] As shown in Table 8.1 below both batches of botanically
derived purified CBD induced a significant increase in neurogenesis
and neuronal differentiation of hNSCs, this is indicated by an
increased number of MAP2-MAP2+ cells after incubation with CBD for
3 or 5 days.
TABLE-US-00006 TABLE 8.1 Percentage MAP2+ cells after 3 and 5 days
incubation Mean % Mean % MAP2+ cells MAP2+ cells Sample (3 days) (5
days) Vehicle 4.59 .+-. 0.3 5.77 .+-. 0.23 Botanically derived
purified 7.06 .+-. 0.11 *** 20.38 .+-. 0.86 *** CBD (batch 1) 0.1
.mu.M Botanically derived purified 28.29 .+-. 1.7 *** 38.38 .+-. 1
*** CBD (batch 1) 1 .mu.M Botanically derived purified 6.71 .+-.
0.91 *** 18.83 .+-. 1.22 *** CBD (batch 2) 0.1 .mu.M Botanically
derived purified 20.04 .+-. 1.37 *** 33.74 .+-. 1.83 *** CBD (batch
2) 1 .mu.M Synthetic CBD 0.1 .mu.M 4.87 .+-. 0.1 6.02 .+-. 0.3
Synthetic CBD 1 .mu.M 6.11 .+-. 0.3 7.09 .+-. 0.1 *** P < 0.0001
vs vehicle-treated cells. Two-way analysis of variance (ANOVA)
followed by the post-hoc Bonferroni test for pairwise
comparison
[0318] Botanically derived purified CBD at a concentration of 1
.mu.M was found to be more potent than the 0.1 .mu.M concentration
for both batches and at both time points (3 days and 5 days of
treatment). Interestingly, the same concentrations of synthetic CBD
did not have a significant effect on neurogenesis or neuronal
differentiation relative to vehicle-treated cells.
Effect of Botanically Derived Purified CBD and Synthetic CBD on
Axonal Outgrowth and Neuronal Differentiation of hNSCs
[0319] This study involved examining the effect of the two test
compounds on neuronal differentiation of hNSCs after 3 or 5 days of
treatment using GAP43 immunocytochemistry. As shown in Table 8.2
below, after 3 and 5 days of treatment with botanically derived
purified CBD significantly increased neurogenesis by enhancing
neurite elongation as confirmed by GAP43 expression in the vast
majority of the cells.
TABLE-US-00007 TABLE 8.2 Percentage GAP43+ cells after 3 and 5 days
incubation Mean % Mean % GAP43+ cells GAP43+ cells Sample (3 days)
(5 days) Untreated 5.96 .+-. 1.05 7.08 .+-. 1.5 Botanically derived
26.39 .+-. 3.40 37.32 .+-. 0.9 purified CBD 1 .mu.M Synthetic CBD 1
.mu.M 8.40 .+-. 2.0 9.15 .+-. 1.6
Conclusion
[0320] Together, these studies indicate that botanically derived
purified CBD, which comprises less than or equal to 2% other
cannabinoids including THC, CBD-C1, CBDV and CBD-C4 are capable of
inducing differentiation and neurite outgrowth.
[0321] Remarkably the synthetic CBD (which does not contain these
other cannabinoids) appears to not have any effect on neurogenesis
and differentiation of hNSCs when supplied at the same
concentration as the botanically derived purified CBD.
[0322] The effect shown by botanically derived purified CBD on
neurogenesis and neurite outgrowth could be important for the
treatment of neurodegenerative disease or brain injuries.
Example 9: Comparison of Botanically Derived Purified CBD and
Synthetic CBD in a Mouse Model of Epilepsy
[0323] This example demonstrates the effect of botanically derived
purified CBD and synthetic CBD on mice in a maximal electroshock
seizure test.
Methods
Animals and Experimental Conditions
[0324] This study was performed using male C57Bl6 mice, weighing
between 23.6-31.2 g, which were purchased from a licensed breeder
(Charles River, UK). Naive mice were acclimatized to the procedure
room in their home cages, with food and water available ad libitum.
Animals were housed in groups of 2-3, in standard caging on a 12
hr/12 hr light-dark cycle. All animals were tail marked and weighed
at the beginning of the study. Animals were randomly assigned to
vehicle or treatment groups.
Anti-Epileptic Drugs (AED)
[0325] The following drugs were used in this study: synthetic CBD
(SYN) and botanically derived purified CBD (BOT). The control
vehicle used was (1:1:18), which was 5% Ethanol, 5% Kolliphor, 90%
saline, and has been used extensively throughout similar seizure
tests. All drugs were dissolved in the control vehicle for
comparison. Animals were dosed (10 ml/kg) i.p. at 60 minutes prior
to the administration of the maximal electroshock seizure (MES)
test. In addition to the vehicle group, the dosage groups used for
each treatment and included doses of 10, 50, 100 (Experiment 1
only), 150 and 200 mg/kg for CBD (SYN) and CBD (BOT). The number of
animals in each of these groups were 10 (n=10).
[0326] Two separate experiments were undertaken using two different
batches of the botanically derived purified CBD. In the first
experiment the concentration of THC in botanically derived purified
CBD was 0.02% THC (w/w) whereas in the second experiment the
concentration of THC in the botanically derived purified CBD was
approximately 0.1% THC (w/w).
Maximal Electroshock Seizure (MES) Test
[0327] The protective activity of CBD (SYN) and CBD (BOT) was
evaluated and expressed as effective doses (ED.sub.50 in mg/kg),
protecting 50% of mice against MES-induced tonic seizures.
Electroconvulsions were produced by a fixed current intensity of 30
mA and corneally delivered electroshock (0.2 second duration). Mice
were individually assessed for seizures following this
pre-determined high level (30 mA) corneally delivered electroshock
of sufficient intensity to reliably produce tonic hind limb
extension seizures in 100% of control animals. Induction of seizure
was measured as an all-or-nothing effect scored as either present
(+) or absent (0) for each animal. Data were collected by an
observer unaware of the treatment for each animal and expressed as
the total number of +s and 0s for each treatment group. Percent
inhibition of relevant vehicle treated group (the degree of
protection relative to vehicle treated controls) was then
generated.
Statistical Analysis
[0328] All statistical tests were performed using commercially
available GraphPad Prism version 7.0 for Windows (GraphPad
Software, San Diego, Calif.). The effective doses (ED.sub.50 in
mg/kg), protecting 50% of mice against MES-induced tonic seizures
for CBD (SYN) and CBD (BOT) were calculated in Prism using
Sigmoidal dose-response--variable slope and log-probit analysis.
The ED.sub.50 values from CBD (SYN and CBD) were compared using the
t test approach to compare two best best-fit values from one
experiment to statistically evaluate the significance of
differences between these.
Results
[0329] The anticonvulsant effects shown by given doses of CBD (SYN)
and CBD (BOT 0.02% THC) were analyzed by sigmoidal curve analyses
as shown in FIG. 13. The ED.sub.50 values were derived from these
sigmoidal curves as CBD (SYN): 77.63 mg/kg and CBD (BOT 0.02% THC):
70.22 mg/kg.
[0330] These ED.sub.50 values were statistically compared using the
t test approach for two best best-fit values from one experiment.
As a result, these ED.sub.50 values were determined statistically
different as shown in Table 9.1 below (p=0.0013).
[0331] The mouse doses were converted to human equivalent dose
(HED) of 10 mg/kg, assuming 60 kg human, and the difference in HED
using CBD (BOT 0.02% THC) over CBD (SYN) is 10.00%.
TABLE-US-00008 TABLE 9.1 Experiment 1 - Analysis from the t-test
approach to compare the ED.sub.50 of CBD (SYN) versus CBD (BOT
0.02% THC) and subsequent dose conversion to human equivalent dose
(HED) CBD CBD CBD CBD (SYN) (BOT) (SYN) (BOT) Diff. ED.sub.50
ED.sub.50 t-test HED - HED - in HED Analysis (mg/kg) (mg/kg) (p
value) mg mg (%) Sigmoidal 77.60 70.20 0.0013 600.0 542.8 10.0
curve - constant top
[0332] The anticonvulsant effects shown by given doses of CBD (SYN)
and CBD (BOT 0.02% THC) were analyzed by sigmoidal curve analyses
as shown in FIG. 14. The ED.sub.50 values were derived from these
sigmoidal curves as CBD (SYN): 77.40 mg/kg and CBD (BOT 0.1% THC):
57.94 mg/kg.
[0333] These ED.sub.50 values were statistically compared using the
t test approach for two best best-fit values from one experiment.
As a result, these ED.sub.50 values were determined statistically
different as described in Table 9.2 (p=0.0000015).
[0334] The mouse doses have been converted to human equivalent dose
(HED) of 10 mg/kg, assuming 60 kg human, and the difference in HED
using CBD (BOT 0.08% THC) over CBD (SYN) is 28.75%.
TABLE-US-00009 TABLE 9.2 Experiment 2 - Analysis from the t-test
approach to compare the ED.sub.50 of CBD (SYN) versus CBD (BOT
approx. 0.1% THC) and subsequent dose conversion to human
equivalent dose (HED) CBD CBD CBD CBD (SYN) (BOT) (SYN) (BOT) Diff.
ED.sub.50 ED.sub.50 t-test HED - HED - in HED Analysis (mg/kg)
(mg/kg) (p value) mg mg (%) Sigmoidal 77.40 57.94 0.0000015 600.0
449.15 28.75 curve - constant top
Conclusion
[0335] The data generated in this study indicates that treatment
with botanically derived purified CBD is more efficacious than
treatment with synthetic CBD in the maximal electroshock model of
epilepsy.
[0336] Such data are significant as they demonstrate that such a
CBD composition may be useful in the treatment of epilepsy.
Example 10: Comparison of Botanically Derived Purified CBD and
Synthetic CBD in an Animal Model of Schizophrenia
[0337] The effect of PCP in the novel object recognition (NOR) test
is a model of visual recognition memory deficits similar to those
observed in schizophrenia. The atypical antipsychotics, clozapine
and risperidone, can attenuate the deficit. The study was designed
to determine if botanically derived purified CBD and/or synthetic
CBD could attenuate the deficits in novel object recognition caused
by administration of PCP.
Methods
[0338] Female hooded-Lister rats were used for this experiment.
Rats were housed in groups of 5 under standard laboratory
conditions under a 12 hr light: dark cycle, with lights on at 0700
hr. Testing was carried out in the light phase. Rats were randomly
assigned to two treatment groups and treated with vehicle, n=20
(distilled water, i.p.) or Phencyclidine hydrochloride (PCP), n=100
(2 mg/kg, i.p. twice daily for 7-days). PCP was dissolved in
distilled water. This was followed by a 7-day wash out period
before the rats were tested following acute treatment with CBD,
risperidone or vehicle.
[0339] Risperidone (0.1 mg/kg) was dissolved in a minimum volume of
acetic acid, made up to volume with distilled water and pH adjusted
to 6 with 0.1M NaOH and administered via the i.p. route in a volume
of 1 ml/kg, 120 minutes prior to testing.
[0340] Botanically derived purified CBD was tested at 2, 10, 20 or
100 mg/kg and was dissolved in 2:1:17 (Ethanol:Cremofor:Saline
0.9%) and administered via the i.p. route in a volume of 5 ml/kg,
60 minutes prior to testing. The THC concentration in the
botanically derived purified CBD was 0.03% (w/w) THC.
[0341] Synthetic CBD at 1, 2, 5, 10 or 20 mg/kg was dissolved in
2:1:17 (Ethanol:Cremofor:Saline 0.9%) and administered via the i.p.
route in a volume of 5 ml/kg, 120 minutes prior to testing.
[0342] Rats were allowed to habituate to the empty test box and the
behavioral test room environment for one hour on day 1. Prior to
behavioral testing on day 2, rats were given a further 3 minute
habituation.
[0343] Following the 3 minute habituation period, the rats are
given two 3 minute trials (T1 and T2) which were separated by a 1
minute inter-trial interval in the home cage during which the
objects were changed. Behavior in all trials was recorded on video
for subsequent blind scoring.
[0344] In the acquisition trial (T1), the animals were allowed to
explore two identical objects (A1 and A2) for 3 minutes. In the
retention trial (T2), the animals were allowed to explore a
familiar object (A) from T1 and a novel object (B) for 3 minutes.
The familiar object presented during T2 was a duplicate of objects
presented in T1 in order to avoid any olfactory trails.
[0345] Object exploration was defined by animals licking, sniffing,
or touching an object with the forepaws while sniffing the object,
but object exploration did not include an animal leaning against,
turning around, standing on or sitting on an object. The
exploration time(s) of each object (A, B, familiar and novel) in
each trial were recorded using two stopwatches and the following
factors were calculated: total exploration time of both objects in
the acquisition trial, total exploration time of both objects in
the retention trial. Habituation of exploratory activity included
the exploration time, as measured by the number of lines crossed,
for both trials.
[0346] All data were assessed for normality using D'Agostino and
Pearson normality test. Data non-normally distributed were analyzed
using Kruskal-Wallis followed by planned comparisons with Dunn's
correction. Normally distributed data were analyzed using one-way
ANOVA followed by planned comparisons with Sidak's correction. All
analyses were carried out using GraphPad Prism V7.03.
Results
[0347] As shown in FIG. 15, botanically derived purified CBD (2-100
mg/kg i.p., 120 min ppt) attenuated a sub-chronic PCP-induced
deficit in novel object recognition in rats (n=9-10 per group),
with a minimal effective dose (MED) of 2 mg/kg, (p=<0.05).
[0348] As shown in FIG. 16, synthetic CBD (2-100 mg/kg i.p., 120
min ppt) attenuated a sub-chronic PCP-induced deficit in novel
object recognition in the rat (n=8-10 per group), with a minimal
effective dose (MED) of 10 mg/kg (p=<0.01).
[0349] Interestingly, the difference in potency (minimal effective
dose) between the synthetic CBD and botanically derived purified
CBD means that in a 70 kg human being dosed at 20 mg/kg/day of CBD
there is a significant difference in the amount of CBD required. As
shown below:
Rat dose of 2 mg/kg/day=(2.times.0.16)=0.32 mg/kg/day in a
human=(0.32.times.70)=22.4 mg/day for highly purified CBD of
botanic origin.
Rat dose of 10 mg/kg/day=(10.times.0.16)=1.6 mg/kg/day in a
human=(1.6.times.70)=112 mg/day for synthetic CBD
Calculations are based on FDA dose conversion guidance from animal
to human (to convert a mg/kg dose in a rat to a mg/kg human
equivalent dose the rat dose is multiplied by 0.16).
Conclusion
[0350] Botanically derived purified CBD has been shown to be useful
in attenuating the sub-chronic PCP-induced deficit in novel object
recognition in rats at a much lower minimal effective dose (MED)
than synthetic CBD suggesting it would be a useful treatment option
in schizophrenia and associated conditions.
[0351] Given the difference in potency, using synthetic CBD in a
human would require five times the amount of CBD than botanically
derived purified CBD. Such a difference in potency is important
given that CBD is an expensive compound to produce whether from a
synthetic or botanic route.
[0352] Example 11: Comparison of High Concentrations of THC in
Combination with Botanically Derived Purified CBD in an Animal
Model of Schizophrenia
[0353] The novel object recognition (NOR) test as described in
Example 9 above is a model of visual recognition memory deficits
similar to those observed in schizophrenia. The study was designed
to determine if higher concentrations of THC, as are commonly found
in vernacular CBD oil preparations would affect the ability of
botanically derived purified CBD to attenuate the deficits in novel
object recognition caused by administration of PCP in rats.
[0354] Vernacular CBD preparations typically comprise between 3 and
20% (w/w) THC.
Methods
[0355] The same method that is described in Example 10 above was
employed in this experiment.
[0356] Botanically derived purified CBD was tested at 20 mg/kg with
an addition of either 10% (w/w) or 20% (w/w) THC added to the CBD.
Botanically derived purified CBD which comprised 0.08% THC was made
up to 10% THC and 20% THC with purified botanically derived
THC.
[0357] The cannabinoids were dissolved in 2:1:17
(Ethanol:Cremofor:Saline 0.9%) and administered via the i.p. route
in a volume of 5 ml/kg, 120 minutes prior to testing.
[0358] The behavioral test was performed as described in Example 10
above.
[0359] All data were assessed for normality using D'Agostino and
Pearson normality test. Data non-normally distributed were analyzed
using Kruskal-Wallis followed by planned comparisons with Dunn's
correction. Normally distributed data were analyzed using one-way
ANOVA followed by planned comparisons with Sidak's correction. All
analyses were carried out using GraphPad Prism V7.03.
Results
[0360] The animals treated with the botanically derived purified
CBD comprising 20% (w/w) THC were found to be sedated and as such
no data was generated with respect to the novel object recognition
for this group.
[0361] Some of the animals treated with the botanically derived
purified CBD comprising 10% (w/w) THC were also found to be sedated
and as such data was only generated with five of the 15 test
animals for this group.
[0362] As can be seen in FIGS. 17 to 19 animals treated with
botanically derived purified CBD at 20 mg/kg were able to attenuate
the sub-chronic PCP-induced deficit in novel object recognition.
Interestingly, it was observed that this effect was removed in
animals that were treated with botanically derived purified CBD
comprising 10% (w/w) THC.
[0363] FIG. 17 demonstrates that botanically derived purified CBD
(20 mg/kg i.p., 120 min ppt) attenuated a sub-chronic PCP-induced
deficit in novel object recognition in rats (n=15 per group),
(p=<0.001). Whereas in the group administered CBD (20 mg/kg)
plus THC (1.984 mg/kg, i.p., 120 min ppt) this effect was lost
(n=5). Effects similar to those observed in the vehicle treated PCP
rats occurred.
[0364] FIG. 18 details the discrimination index (DI) in rats
treated with botanically derived purified CBD supplemented with THC
at 10 and 20% (w/w). Rats treated with botanically derived purified
CBD (20 mg/kg i.p., 120 min ppt) had a DI which was similar to
those rats untreated with PCP (n=15 per group), (p=<0.01).
Whereas in the group administered CBD (20 mg/kg) plus THC (1.984
mg/kg, i.p., 120 min ppt) this effect was lost (n=5).
[0365] FIG. 19 details the number of line crossings in rats treated
with botanically derived purified CBD supplemented with THC at 10
and 20% (w/w). Rats treated with CBD (20 mg/kg) plus THC (1.984
mg/kg, i.p., 120 min ppt) had a reduced number of line crossings
(n=5).
Conclusion
[0366] As such it would appear that a small concentration of THC as
is found in the botanically derived purified CBD is efficacious
however increasing the concentration of THC to levels found in
vernacular CBD preparations is detrimental to cognitive and social
deficit.
Conclusion of Data Demonstrating Therapeutic Efficacy of
Botanically Derived Purified CBD
[0367] The data presented in Examples 8 to 11 above describe the
pharmacological properties of the botanically derived purified CBD
preparation that was produced from a high-CBD plant in comparison
to synthetic CBD.
[0368] As we have determined in Examples 2 to 7 the botanically
derived purified CBD used in these experiments has a precise
composition which differs from that of synthetic CBD albeit the two
compositions have the same concentration of CBD.
[0369] These data demonstrate in three different models that the
botanically derived purified CBD is more efficacious than synthetic
CBD. These data are surprising given that the difference in the
composition between the two types of CBD tested lies only within
what may be considered to be the impurities.
[0370] Such data are important in many respects. Firstly, when
treating a patient with a disease or condition the medical
practitioner is desirous of ensuring the most effective treatment
possible. These data show that the botanically derived purified CBD
is more effective than synthetic CBD and as such is a more valuable
treatment option, particularly in patients that are suffering from
difficult to treat conditions such as many of the epilepsy
syndromes such as Dravet syndrome or Lennox-Gastaust syndrome.
[0371] Secondly, the findings from these experiments have
demonstrated that botanically derived purified CBD is effective at
a lower minimal effective dose than synthetic CBD. As such when
administering the botanically derived purified CBD a smaller amount
of the composition may be provided. This has many benefits
including a lower cost of goods, potentially lower associated side
effects and patient compliance.
[0372] Furthermore, these data also demonstrate that using an
unrefined source of CBD such as that which is found from CBD oil
retailers does not produce the same efficacious benefits as is
observed with botanically derived purified CBD. The detrimental
effects shown on cognition when the amount of THC in the CBD was
increased to vernacular levels suggests that such compositions
would not be suitable for the treatment of many diseases and
conditions.
[0373] In summary, the precise composition of components within the
botanically derived purified CBD are of great benefit. Such a
composition may be reproduced using either CBD of botanical origin
or components that are produced synthetically.
* * * * *