Cannabidiol-based formulation and its use for treating pain, inflammation

Abstract

The invention relates to cannabidiol-based (CBD) formulations, and to their use in treating, preventing or alleviating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea in a subject. The invention also concerns low-dose CBD compositions, and extends to their use in methods of treatment.

Description

[0001] Cannabidiol-based Formulation

[0002] The present invention relates to cannabidiol-based (CBD) formulations, and particularly, although not exclusively, to their use in treating, preventing or alleviating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea in a subject. The invention is especially concerned with low-dose CBD compositions, and extends to their use in methods of treatment.

[0003] The cannabinoids are several structural classes of compounds found primarily in the cannabis plant. The best studied cannabinoids include cannabidiol (CBD), tetrahydrocannabinal (THC) and cannabinol (CBN), with CBD being the most widely investigated phytocannabinoid (i.e., a plant-derived cannabinoid) due to its therapeutic activities and lack of psychoactive effects.

[0004] Unfortunately, the CBD that is used for the treatment of various health conditions is usually extracted directly from the cannabis plant, and it is very difficult, if not impossible, to standardise CBD extracts due to a wide variety of factors influencing the biochemical composition of the cannabis plant, such as the geographical location of where the cannabis plant is grown, the fluctuating growth conditions of the plant (i.e. soil and weather conditions), the time of year of CBD extraction from the plant, and differences in extraction methods from the plant, etc. This significant variability in the cannabis plant and extraction parameters results in the CBD formulations that are currently available suffering from the problem of not being consistently reproducible and so not always having the correct amounts of CBD that is listed on the label (Liebling et al., 2022, An Analysis of Over-the-Counter Cannabidiol Products in the United Kingdom, Cannabis and Cannabinoid Research, 7, 2, 207-213).

[0005] Formulations comprising cannabinoids are referred to as a Cannabis-Based Product for Medicinal use (CBPM). CBPMs have been added to the treatment paradigm for many patients suffering from acute and chronic inflammatory conditions, pain, and symptoms of negative affect, such as anxiety and depression that have limited or a lack of response to traditional therapeutic approaches. Due to this, the National Institute for Health and Care Excellence (NICE) and the International Association for the Study of Pain (IASP) have made specific calls for research using CBPMs in humans. However, there is still an unmet need for treating autoimmune diseases, multiple treatment-resistant conditions, such as fibromyalgia, chemotherapy-induced peripheral neuropathy (CIPN) and endometriosis, which are characterised by pain, inflammation and other negative effects that have an adverse influence on the quality of life of patients suffering from these conditions.

[0006] Accordingly, there is a need for a new medical CBPM formulation for the treatment of various conditions, such as pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea, and which is produced under defined pharmaceutical standards and under the guidelines of GMP for use in medical research and practice, and in foodstuffs, and which therefore facilitates the access and delivery of a pharmaceutical standard CBPM to clinical trials and to benefit patients. There is also a need to produce standardised CBD-based formulations for treating these conditions which contain reproducibly consistent levels of CBD while lacking any significant levels of THC. This standardisation means that the composition and biological activity of the CBD formulation will be the same between batches, giving patients significantly more confidence in the CBPM product and its therapeutic effects for treating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea.

[0007] As described in the Examples, and as shown in Figures 1 and 2, the inventors have devised an innovative method for producing a highly standardised CBD-based formulation that can be reproducibly delivered at an accurate low dose, for the treatment of pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea. The formulation comprises the cannabinoid, CBD, using an isolate of CBD of known purity allowing the preparation of a formulation with standardised amounts of CBD, whilst simultaneously minimising any contamination with THC. As described in Example 13, the inventors have now demonstrated that surprisingly low doses of CBD (60mg / day or 70mg / day; or 0.8mg / kg / day or Img / kg / day CBD) is effective for reducing pain in less than 4 weeks of therapy, such as that caused by chemotherapy-induced peripheral neuropathy (CIPN).

[0008] Thus, according to a first aspect of the invention, there is provided a composition comprising cannabidiol (CBD), for use in treating, preventing or alleviating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea in a subject, wherein a dosage of between O.lmg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0009] In a second aspect, there is provided a method of treating, preventing or ameliorating a subject suffering from pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea, the method comprising administering, or having administered, to a subject in need of such treatment, between O.lmg and 20mg CBD per kilogram subject body weight per day.

[0010] Unexpectedly, low doses of CBD have been shown to be surprisingly effective for treating the above conditions, and in pain in less than 4 weeks of therapy, such as that caused CIPN. Advantageously, and preferably, the invention combines CBD of a known purity to produce a fully standardized formulation with a known concentration and purity of each of the constituents, which is fully reproducible, with no variability between different batches, and which can therefore be administered reproducibly at low doses. In one embodiment, the CBD-based composition of the invention is referred to herein as MRX1, embodiments of which are shown in Tables 2, 3 and 4. Accordingly, a certificate of analysis can be given for each batch of the composition of the invention, showing its stability data, as well as evidence that there is substantially no contamination with any THC, thereby providing the patient with full confidence of the composition and, therefore, its therapeutic efficacy.

[0011] In an embodiment, between O.lmg and 18mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 16mg CBD per kilogram subject body weight per day is administered to the subject.

[0012] In another embodiment, between O.lmg and 14mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 12mg CBD per kilogram subject body weight per day is administered to the subject.

[0013] In another embodiment, between O.lmg and llmg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and lOmg CBD per kilogram subject body weight per day is administered to the subject.

[0014] In another embodiment, between O.lmg and 9mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 8mg CBD per kilogram subject body weight per day is administered to the subject.

[0015] In another embodiment, between O.lmg and 7mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 6mg CBD per kilogram subject body weight per day is administered to the subject. In another embodiment, between O.lmg and 5mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 4mg CBD per kilogram subject body weight per day is administered to the subject.

[0016] In another embodiment, between O.lmg and 3mg CBD per kilogram subject body weight per day is administered to the subject, or between O.lmg and 2mg CBD per kilogram subject body weight per day is administered to the subject.

[0017] In a typical embodiment, between O.lmg and Img CBD per kilogram subject body weight per day is administered to the subject.

[0018] Any of the above upper and lower values may be combined in any combination.

[0019] In an embodiment, between 0.15mg and 20mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.2mg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0020] In another embodiment, between 0.3mg and 20mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.4mg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0021] In another embodiment, between 0.5mg and 20mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.6mg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0022] In another embodiment, between 0.7mg and 20mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.8mg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0023] In another embodiment, between 0.9mg and 20mg CBD per kilogram subject body weight per day is administered to the subject, or between l.Omg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

[0024] Any of the above upper and lower values may be combined in any combination. In an embodiment, between 0.15mg and 4.5mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.2mg and 4mg CBD per kilogram subject body weight per day is administered to the subject.

[0025] In another embodiment, between 0.3mg and 3.5mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.4mg and 3mg CBD per kilogram subject body weight per day is administered to the subject.

[0026] In another embodiment, between 0.45mg and 2.5mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.50mg and 2mg CBD per kilogram subject body weight per day is administered to the subject.

[0027] In another embodiment, between 0.55mg and 1.75mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.60mg and 1.5mg CBD per kilogram subject body weight per day is administered to the subject.

[0028] In another embodiment, between 0.65mg and 1.3mg CBD per kilogram subject body weight per day is administered to the subject, or between 0.7mg and l.lmg CBD per kilogram subject body weight per day is administered to the subject.

[0029] Typically, between 0.8mg and Img CBD per kilogram subject body weight per day is administered to the subject.

[0030] Typically, about 0.8mg CBD per kilogram subject body weight per day is administered to the subject.

[0031] Typically, about Img CBD per kilogram subject body weight per day is administered to the subject.

[0032] Any of the above upper and lower values may be combined in any combination.

[0033] In one embodiment, the composition and therefore the above doses may be administered to the subject as a single dose per day.

[0034] In another embodiment, however, the CBD doses may be administered to the subject as two or more doses per day. Typically, two doses are administered per day. Therefore, for example, in an embodiment in which 0.8mg CBD / kg body weight / day is administered, then when given as two separate doses, 0.4mg CBD / kg body weight bis in die (b.i.d.) is administered.

[0035] Typically, for example, in an embodiment in which Img CBD / kg body weight / day is administered, then when given as two separate doses, about 0.5mg CBD / kg body weight bis in die (b.i.d.) is administered.

[0036] As such, in some embodiments, between 0.2mg and lOmg CBD / kg body weight bis in die (b.i.d.) is administered, or between 0.4mg and 5mg CBD / kg body weight bis in die (b.i.d.) is administered.

[0037] In an embodiment, between 30 and lOOmg CBD per day is administered to the subject, or between 40 and 90mg CBD per day is administered to the subject.

[0038] In another embodiment, between 50 and 80mg CBD per day is administered to the subject, or between 55 and 75mg CBD per day is administered to the subject.

[0039] Typically, between 60 and 70mg CBD per day is administered to the subject.

[0040] As mentioned above, the CBD may be given as two or more doses per day.

[0041] Therefore, for example, in an embodiment in which 60mg CBD / day is administered, then when given as two separate doses, 30mg CBD bis in die (b.i.d.) is administered. In an embodiment in which 70mg CBD / day is administered, then when given as two separate doses, 35mg CBD bis in die (b.i.d.) is administered.

[0042] In some embodiments, the pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea are selected from the group of conditions consisting of: chemotherapy-induced peripheral neuropathy (CIPN), endometriosis, osteoarthritis, rheumatoid arthritis, ulcerative colitis, Crohn's disease, fibromyalgia, irritable bowel syndrome, asthma, chronic obstructive pulmonary disease (COPD), gout, scleroderma, lupus, Ehlers-Danlos syndrome, pericarditis, myocarditis, myocardial infarction, chemotherapy-induced nausea, anxiety, Type I diabetes, cardiovascular disease, primary hypertension, secondary hypertension, resistant hypertension, isolated systolic hypertension, malignant hypertension, obstructive hypertrophic cardiomyopathy, nonobstructive hypertrophic cardiomyopathy, pulmonary fibrosis, liver fibrosis, heart fibrosis, kidney fibrosis, mediastinal fibrosis, retroperitoneal cavity fibrosis, bone marrow fibrosis, skin fibrosis, and scleroderma.

[0043] Typically, the composition is used to treat, prevent or alleviate pain.

[0044] More typically, the composition is used to treat, prevent or alleviate chemotherapy-induced peripheral neuropathy (CIPN).

[0045] The cannabinoid-based composition of the invention is typically in a liquid form. The formulation may, for example, comprise a tincture.

[0046] In an embodiment, the formulation is administrable by oral, sublabial, buccal, sublingual, or oropharyngeal administration delivery. The preparation of the liquid formulations for sublabial, buccal, oropharyngeal or sublingual (oral) delivery allows the introduction of the active agents (i.e., the CBD) into the systemic circulation via a mucous membrane, increasing the onset of activity and potency due to bypassing first-pass metabolism and increasing the drug bioavailability. Advantageously, given the lipophilicity and poor bioavailability of oral administration of cannabinoids for gastrointestinal absorption, which is considered the most popular method currently in the market for the delivery of medicines, the formulations of the invention surprisingly enable administration by this route. The liquid formulations may also be prepared for inhaled or nebulised delivery to the respiratory tract, as well as formulations for topical application to the skin and mucosal surfaces.

[0047] The composition may be administered at least once a day, at least twice a day, at least once a week, or at least once a month. The formulation may be administered two or three times a day, two or three times a week, or two or three times a month.

[0048] The composition may be administered to a "subject" in need of the treatment, which may be a vertebrate, mammal, or domestic animal. Hence, the compositions according to the invention may be used to treat any mammal, for example livestock (e.g., a horse), domestic pets, or may be used in other veterinary applications. Most typically, however, the subject is a human being. A "therapeutically effective amount" of the composition is any amount which, when administered to the subject, is the amount of the aforementioned that is needed to treat the target medical condition.

[0049] The chemical structure of the phytocannabinoid, CBD, comprises Formula [I]

[0050]

[0051] Typically, the CBD comprises an isolate of CBD. The purity of the CBD may be at least 97%, at least 98% or at least 99%, or about 98-100%, as measured by HPLC-UV assay at 220nm detection wavelength. As shown in Figure 3, the inventors have a certificate of analysis (COA) for the CBD isolate. The specification has been set in order to comply with ICH Q3A guidance on impurities in new Drug Substance, i.e., to demonstrate sufficient control of Organic impurities (minor cannabinoids, solvents etc.), inorganic impurities (e.g., heavy metals) and microbiological contamination. Particularly relevant is the decision to set the specification for d9-tetrahydrocannabinol to 0.02%, which is lower than the requirements of less than, or equal to, 0.15% for any single known organic impurity.

[0052] The concentration of the CBD in the cannabinoid-based formulation may be at least 2% (w / v), at least 3% (w / v) or at least 4% (w / v). The concentration of the CBD in the cannabinoid-based formulation may be at least 5% (w / v), at least 6% (w / v) or at least 7% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation may be at least 8% (w / v), at least 9% (w / v) or at least 10% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation may be at least 11% (w / v), or at least 12% (w / v).

[0053] The concentration of the CBD in the cannabinoid-based formulation may be less than 20% (w / v), less than 19% (w / v) or less than 18% (w / v). The concentration of the CBD in the cannabinoid-based formulation may be less than 17% (w / v), less than 16% (w / v) or less than 15% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation may be less than 14% (w / v), less than 13% (w / v) or less than 12% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation may be less than 11% (w / v), less than 10% (w / v) or less than 9% (w / v).

[0054] The concentration of the CBD in the cannabinoid-based formulation may be between 1 and 25% (w / v), between 3 and 20% (w / v) or between 5 and 17% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation is between 7 and 13% (w / v), between 8 and 12% (w / v) or between 9 and 11% (w / v). Typically, the concentration of the CBD in the cannabinoid-based formulation is about 10% (w / v). It will be appreciated that any of the lower values for the concentration of the CBD mentioned herein may be combined with any of the upper values for the concentration of the CBD.

[0055] Typically, the composition does not comprise cannabichromene (CBC). Typically, the composition does not comprise cannabigerol (CBG). Typically, the composition does not comprise cannabigerol monomethyl ether (CBGM).

[0056] Typically, the formulation substantially lacks any THC. THC can be detected using any standard analytical chemistry approaches, such as liquid chromatography with ultraviolet (UV) or mass spectrometric (MS or MS / MS) detection, e.g. Lukas Vaclavik et al., (2019) 'Quantitation of Cannabinoids in Cannabis Dried Plant Materials, Concentrates, and Oils Using Liquid Chromatography-Diode Array Detection Technique with Optional Mass Spectrometric Detection: Single-Laboratory Validation Study, First Action 2018.11' Journal of AOAC INTERNATIONAL, Volume 102, Issue 6, 1 November 2019, Pages 1822-1833.

[0057] The concentration of THC in the composition is ideally less than lOOOpg per finished product, e.g. for 30ml this equates to 0.0033% (w / v). Therefore, typically the concentration of THC in the composition is less than 0.01% (w / v), more typically less than 0.005% (w / v), and even more typically less than 0.001% (w / v).

[0058] Typically, the concentration of THC in the composition is less 0.0005% (w / v), and even more typically less than 0.0002% (w / v), and most typically less than 0.0001% (w / v). Typically, however, the composition comprises no detectable THC using reasonable validated means of analysis. In an embodiment, the composition may further comprise one or more terpene. The one or more terpen may be selected from a group of terpenes consisting of: myrcene; caryophyllene; ocimene; pinene; limonene; humulene; and linalool.

[0059] Terpenes are lipids (complex compounds of fatty acids) and belong to the terpenoid family. Chemically, terpenes present a distinctive carbon skeleton which consists of a basic five-carbon isoprene unit (CsHs, 2-methyl-l,3-butadiene) linked together most commonly in a head-to-tail arrangement. However, they can be built in other configurations with varying degrees of oxidation, unsaturation, functional groups and ring closures, leading to a vast diversity of structural classes. Nevertheless, terpenes can be grouped into categories according to the number of isoprene units (n) in the molecule. Isoprene is a gaseous hydrocarbon released by multiple plants as a natural by-product of plant metabolism. However, some of the larger and more complex terpenes, such as squalene and lanosterol, are also found in animals.

[0060] In a some embodiments, the cannabinoid-based composition further comprises a pharmaceutically acceptable excipient or carrier.

[0061] A "pharmaceutically acceptable vehicle" as referred to herein, is any known compound or combination of known compounds that are known to those skilled in the art to be useful in formulating pharmaceutical compositions.

[0062] In one embodiment, the pharmaceutically acceptable vehicle may be a liquid, and the pharmaceutical composition is in the form of a suspension in solution. Liquid vehicles are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The CBD may be dissolved or suspended in a pharmaceutically acceptable liquid vehicle such as water, alcohol, ionic buffered solution, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid vehicle can contain other suitable pharmaceutical additives such as solubilisers, emulsifiers, buffers, preservatives, sweeteners, flavouring agents, suspending agents, thickening agents, colours, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid vehicles for oral administration include water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil). Pharmaceutically acceptable carriers, excipients, and diluents are relatively inert substances that facilitate administration or a pharmaceutically effective substance and can be supplied as a liquid solutions or suspensions, as emulsions, or as solid forms suitable for dissolution or suspension in liquid prior to use. For example, an excipient can give forms suitable for consistency, or act as a diluent. Suitable excipients include, but are not limited to stabilizing agents, wetting and emulsifying agents, salts for varying osmolarity, encapsulating agents, pH buffering substances, and buffers. Such excipients include any pharmaceutical agent suitable for direct delivery to the subject (for example sub-lingually) which may be administered without undue toxicity. Pharmaceutical acceptable excipients include, but are not limited to, sorbitol, any of the various TWEEN compounds, and liquids such as water, saline, glycerol and ethanol. Pharmaceutically acceptable salts can be included therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulphates and the like; and the salts of organic acids such as acetates, propionates, malonates, or benzoates.

[0063] In some embodiments, pharmaceutical acceptable excipients may include pharmaceutical acceptable carriers. Such pharmaceutically acceptable carriers can be sterile liquids, such as water and oil, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, sesame oil, or mineral oil. Saline solutions and aqueous dextrose, polyethylene glycol (PEG) and glycerol solutions can also be employed as liquid carriers, particularly for sub-lingual solutions. Additional ingredients may also be used, for example preservatives, buffers, tonicity agents, antioxidants and stabilizers, non-ionic wetting or clarifying agents, or viscosity-increasing agents. A thorough discussion of pharmaceutical acceptable excipients and carriers is available in Remington's Pharmaceutical Sciences (Ed Remington JP and Gennaro AR; Mack Pub. Co. Easton, Pa 1990).

[0064] The pharmaceutically acceptable excipient or carrier may comprise a medium-chain triglyceride (MCT), preferably MCT oil. Suitable MCTs may be selected from a group consisting of a triglycerol linker with three fatty acid residues with carbon chain lengths ranging from 1 to 14. The chemical formula of each of these MCTs is shown below: <>

[0065]

[0066] where X= 1 - 14.

[0067] The MCT oil is preferably sourced from fractionated coconut oil. Typical triglycerides comprise a carbon chain length or C6, C8, CIO or C12. Typically, the triglyceride is almost all C8, which is highly refined.

[0068] All of the features described herein (including any accompanying claims, abstract and drawings), and / or all of the steps of any method or process so disclosed, may be combined with any of the above aspects in any combination, except combinations where at least some of such features and / or steps are mutually exclusive.

[0069] For a better understanding of the invention, and to show how embodiments of the same may be carried into effect, reference will now be made, by way of example, to the accompanying Figures, in which:-

[0070] Figure 1 is a flow diagram showing a method for producing a CBD formulation of the invention comprising a CBD isolate, terpene isolates and an MCT oil, as excipient.

[0071] Figure 2 is a flow diagram showing another method for producing a CBD formulation of the invention comprising a CBD isolate, terpene isolates and an MCT oil, as excipient.

[0072] Figure 3 shows a Certificate of Analysis (COA) for one embodiment of the CBD raw material used in low dosages according to the invention.

[0073] Figure 4 shows a Certificate of Analysis (COA) that the CBD formulation of the invention (referred to as "MRX1") substantially lacks 13 non-CBD cannabinoid analytes.

[0074] Figure 5 shows the cumulative concentration of CBD permeation across porcine mucosal tissue membrane over time.

[0075] Figure 6 shows the design a Phase II placebo controlled double-blind cross-over trial of the low dose CBD-based formulation of the invention for treating Chemotherapy Induced Peripheral Neuropathy (CIPN).

[0076]

[0077] The inventors have devised a method for reproducibly and consistently producing a highly standardised formulation comprising CBD, which can be administered reproducibly at a low dose, for use in treating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea in a subject. The composition lacks detectable levels of THC.

[0078] Example 1 - Method of producing the CBD formulation

[0079] Referring to Figures 1 and 2, the inventors have designed a method for producing a highly uniform and reproducible CBD-based formulation (referred to herein as MRX1). The method involves the following steps:

[0080] 1. Extracting CBD from a Cannabis sativa plant to produce a CBD extract; 2. Purifying a CBD isolate (at a defined purity) from the CBD extract;

[0081] 3. Extracting terpenes from the appropriate species of plant to produce a terpene extract (i.e. an essential oil);

[0082] 4. Purifying a terpene (at a defined purity) from the terpene extract, several of which are then blended together to create a terpene blend;

[0083] 5. Preparing an MCT oil (at a defined purity);

[0084] 6. Mixing together the CBD isolate, the terpene blend and the MCT oil to produce the CBD formulation of the invention.

[0085] Each of these steps will now be described in detail.

[0086] Example 2 - Preparing the CBD isolate

[0087] The inventors have used two different embodiments of a method for extracting the pure CBD isolate, i.e., solvent extraction (shown in Figure 1) and enzyme extraction (shown in Figure 2). Solvent Extraction (Figure 1 )

[0088] Referring to Figure 1 (left-hand side), in a first embodiment, the CBD isolate is prepared with the use of solvent extraction (e.g., using either an alcohol or an alkane) from suitable plant material Cannabis sativa inflorescence and / or leaves), producing a slurry which is then filtered to produce a crude liquid extract. The crude extract is chilled to preferably -80°C for 24 hours in a process called "Winterisation" in order to precipitate fats and waxes that are subsequently filtered off to produce a so-called 'winterised' extract.

[0089] The solvent (i.e., alcohol or alkane) is then removed under reduced pressure and the remaining crude oil 'decarboxylated' to convert the acidic compounds to neutral forms, and this is typically performed at 120°C for at least 30 minutes, ideally at least one hour. The resulting decarboxylated extract is further purified by short path distillation at 130°C-180°C to remove residual extraneous non-cannabinoid materials. The resulting material is dissolved in alcohol (i.e., a solvent) to around the saturation point of the solution, before an alkane (acting as an anti-solvent) is slowly added to induce crystallisation. Upon completion of the first crystallisation (~95% purity CBD), the solid material is filtered off and re-dissolved in alcohol, and then a second crystallisation is performed to produce isolated CBD of the required specification.

[0090] Enzyme extraction (Figure 2)

[0091] With reference to Figure 2, in a second embodiment, the one or more cannabinoid is prepared by using enzyme digestion of plant material (e.g., inflorescence and / or leaves of Cannabis sativa). The plant material can comprise a moisture content of about 40%. The plant material is contacted with an enzyme solution which comprises a combination of one or more of the following enzymes: cellulase, betaglucosidase, hemicellulase, xylanase, glucanase, beta- glucanase, pectinase, amylase, alpha-amylase, phospholipase, beta-mannanase, arabinanase, phytase and protease. The pH of the enzyme solution is adjusted to about pH5.6 using citric acid. A weight ratio of 1-10% (wt) of plant material is used to 99-10% (wt) enzyme solution. The plant material is macerated before, during and / or after contacting with the enzyme solution. In addition to the enzyme solution, the plant material is also contacted with a lipid for extracting the cannabinoid (i.e., CBD). For example, the lipid is vegetable oil, MCT oil, seed oil or olive oil. The weight ratio of lipid to plant material is about 1:1.5. The plant material is also macerated before, during and / or after contacting with the lipid. The mixture of plant material, enzymes and lipid is then stirred and macerated at 10°C-50°C for about an hour. Upon full breakdown and dissolution of the plant material, the oil and water phases are then separated by gravity, centrifugation, filtration or a combination thereof. The oil layer is transferred to a separate vessel having agitation capability. The oil layer is contacted with an aqueous base, such as 0.1M NaOH or KOH, at a pH greater than or equal to 12. The weight of aqueous basic solution relative to the cannabinoid-containing oil is 1:5. This mixture is agitated for at least 20 minutes at a temperature of 40°C-80°C.

[0092] On completion of the previous step, the mixture is contacted with a combination of: (i) sodium chloride and / or calcium carbonate; and (ii) glucose and / or fructose, and then agitated for a further 20 minutes. Upon completion, the layers are separated by gravity or centrifugation, and the aqueous layer may then be separated off. The pH of the separated aqueous layer is reduced to a pH of about 1 by contacting the aqueous layer with 85% ortho-phosphoric acid. The solution is agitated for a further 20 minutes, causing precipitation of cannabinoid acid salts. This liquid is filtered, giving a cannabinoid-rich solid precipitate between 65% - 90% total cannabinoid content. The cannabinoid-rich precipitate is heated to at least 120°C for 60 mins, or until such time that complete decarboxylation has been achieved. The resulting material is dissolved in a solvent (e.g., alcohol) to around the saturation point of the solution, before an anti-solvent (e.g., alkane) is (slowly) added to induce crystallisation. Optionally, the solid material can be filtered off and re-dissolved in solvent (e.g., alcohol), and a second crystallisation may be performed to produce an isolated cannabinoid (preferably, CBD) of the required specification.

[0093] Certificate of Analysis (COA)

[0094] Referring to Figure 3, there is shown a Certificate of Analysis (COA) for one embodiment of the CBD raw material. As can be seen, a wide number of parameters have been measured, and are well within accepted limits. In particular, the inventors note the ability to set the specification for d9-tetrahydrocannabinol (THC) to 0.02%, which is significantly lower than the pharmaceutical requirements of less than, or equal to, 0.15% for any single known organic impurity.

[0095] 3 - Preparing the terpene blend

[0096] Referring to Figures 1 and 2 (right-hand side), the terpenes are prepared by_steam distillation to produce an essential oil from the appropriate species of plant, such as lavender. The resulting essential oil is purified to individual components by flash chromatography with silica. The following example is for the purification of linalool from lavender, though it will be appreciated that other essential oils comprising one or more terpene can be prepared from other plant species.

[0097] The steam distillation equipment is a closed system consisting of a boiling vessel into which macerated lavender is placed, adequate water is then added to allow for mobilisation of the plant material. Steam is introduced into the bottom of the vessel via a dip tube running from the top of the boiling vessel. The mixture is heated to boiling point and the vapour travels through a manifold at the top of the boiling vessel to a water-cooled condenser. The condensate is collected and allowed to separate, where the top essential oil layer is collected.

[0098] The essential oil is dissolved at the appropriate concentration in 90:10 heptane:ethyl acetate and introduced to a flash chromatography system and separated using a silica packed column and 90:10 heptane:ethyl acetate mobile phase. The later fractions yield >95% purity linalool.

[0099] The inventors used isolated terpenes after the purification process as raw materials for preparation of the CBD formulation. The purity standard of the raw terpenes is 95% purity and above (measured by GC). Impurities are mostly other terpenes, and all impurities above 0.2% are identified in order to be included as part of the formulation and to ensure the highest consistency.

[0100] The terpenes are then combined by weight to produce a highly reproducible terpene mixture or blend. As shown in Figures 1 and 2, the resulting essential oils, each comprising a terpene at a 95% purity or above, are mixed together to form a terpene blend, i.e., terpene X, terpene Y and / or terpene Z, and so on), each at a purity of 95% or more. Table 1 shows one exemplary embodiment of the terpene blend.

[0101] Table 1 - Terpene blend

[0102]

[0103]

[0104] Example 4 - Preparing the MCT oil

[0105] Referring to Figures 1 and 2 (left-hand side), a commercially available pharmaceutical grade MCT oil is obtained from H Plus Limited, Walker House, Exchange Flags, Liverpool, L2 3YL, England. For example, the MCT can be sourced from fractionated coconut oil, and has a carbon chain length of C6, C8, CIO or C12, though a highly refined C8 chain length is preferred.

[0106] Example 5 - CBD formulation

[0107] The method comprises combining the CBD isolate from Example 2, the terpene isolate from Example 3, and the MCT oil from Example 4 to produce the cannabinoid-based formulation of the invention. This is achieved by first mixing the CBD isolate with the MCT oil with agitation at about 20°C-50°C. Then, the CBD / MCT solution is mixed with the plurality of blended terpenes with agitation at about 20°C-50°C to create the CBD / MCT / terpene formulation.

[0108] The inventors have designed an embodiment of the CBD-based formulation with the composition shown in Table 2.

[0109] Table 2 - A first embodiment of a CBD Formulation

[0110]

[0111]

[0112] The inventors have designed another embodiment of the CBD-based formulation with the composition shown in Table 3.

[0113] Table 3 - A second embodiment of a CBD Formulation

[0114]

[0115] The inventors have designed another embodiment of the CBD-based formulation with the composition shown in Table 4.

[0116] Table 4 - A third embodiment of a CBD Formulation

[0117]

[0118]

[0119] Example 6 - CBD-onlv formulation (MRX1)

[0120] Analysis of the cannabinoid-based formulation of the invention, referred to herein as MRX1, was performed by Phytovista Laboratories, a specialist ISO accredited laboratory which test CBD & hemp products. Using HPLC-DAD and UV-spectrophotometry, Phytovista Laboratories tested the MRX1 for the presence of 13 non-CBD cannabinoid analytes, namely Cannabidiolic acid (CBDA), Cannabidivarinic acid (CBDVA), Cannabigerol (CBG), Cannabigerolic acid (CBGA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), Cannabicyclol (CBL), Tetrahydrocannabivarinic acid (THCVA), Tetrahydrocannabivarin (THCV), Cannabinol (CBN), A9-Tetrahydrocannabinol (A9-THC), A8-Tetrahydrocannabinol (A8-THC), A9-Tetrahydrocannabinolic acid A (A9-THCA-A).

[0121] As shown in Figure 4, Phytovista Laboratories issued a certificate of analysis confirming that MRX1 contained less than the limit of quantification (<LOQ), where the LOQ was 0.0025, for all 13 analytes.

[0122] Example 7 - MRX1 Bioavailabilitv

[0123] A mucosal skin permeation study of the cannabinoid-based formulation of the invention, referred to herein as MRX1, via Franze cell analysis was performed by Nottingham Trent University.

[0124] Aim

[0125] The aim of the study was to determine the systemic exposure, over time, of MRX1 in a porcine (pig) mucosal tissue permeation study, using an in vitro model (Franz cell apparatus) utilising a proprietary transdermal formulation, i.e., an embodiment of the cannabinoid-based formulation of the invention, containing cannabinoid isolate cannabidiol (CBD) in a lipid matrix (comprising of combinations of the following: medium chain triglycerides (MCT), propylene glycol, ethanol and terpenes (B-Myrcene, B-Caryophyllene, B-Ocimene, o-Pinene, Limonene, o-Humulene, and linalool). Materials and instrumentation

[0126] MRX1, Cannabidiol (CBD) isolate, Kollisolv MCT70, propylene glycol, and anhydrous ethanol were supplied to the University. The cannabidiol reference standard was used as received by the supplier (Restek, # 34011). All other solvents were used as supplied (Analytical or LCMS grade; Fisher Scientific and Sigma) without further purification. Apparatus used included DixonScience Franz cell apparatus (# XFCDR01), and Agilent 7890A series Gas Chromatography coupled with Agilent 5975C MDS with ALS 7693 supported with NIST library search.

[0127] Method

[0128] Franz cell protocol

[0129] Franz cell experiments were carried out on the single MRX1 formulation (0.5 mL sample containing CBD 10-20 % w / v) in triplicate for five sperate formulations, namely BCMS_032_020, BCMS_032_021, BCMS_032_022, BCMS_032_023, and BCMS_032_024. Each Franz cell was carried out over a five-hour period with specific sampling time points at time = 0, 30, 60, 90, 120, 150, 180, 240 and 300 minutes using porcine mucosa tissue membranes.

[0130] In summary, the following procedure was carried out:

[0131] The porcine mucosa tissue membrane tissue was harvested from fresh

[0132] (<4 hours) euthanised pigs (ca. 6 month old) before starting the procedure, ensuring to trim any excess tissue from the skin sample. All samples were washed and dried before loading into the Franz cell apparatus. Franz cell apparatus with a receptor fill volume of ca. 2.0 mL and an effective diffusional area of 1.33 cm2were used.

[0133] The receptor compartment of the Franz cell was filled (ca. 2.0 mL) with degassed phosphate-buffered saline solution (PBS - sodium chloride 8.0

[0134] g / L, potassium chloride 0.2 g / L, disodium hydrogen phosphate 1.15 g / L, potassium dihydrogen phosphate 0.2 g / L) with bovine serum albumin (4% v / v) at pH 7.4 (± 0.1), maintained at 37°C (± 1°C).

[0135] The prepared tissue was placed into the Franz cell apparatus and clamped into position, ensuring a firm seal. The skin was saturated in situ by placing a small amount (ca. 0.5 mL) of buffer medium on top on the skin / donor chamber for prior starting the analysis (1 hr). After ensuring the sample

[0136] was saturated and there were no leaks in the system, buffer medium was removed from the donor chamber.

[0137] The Franz cell apparatus was continually checked to ensure there were no air bubbles present under the skin membrane; if present the sample chamber was carefully angled to remove them. The temperature of the system was maintained at 37°C (± 1°C) with 5% CO2 and kept under constant stirring using a magnetic stirrer bar to ensure homogeneous mixing of the receptor solution throughout the analysis.

[0138] The five suppled formulations for testing were individually applied (0.5 mL, stored at room temperature) to the receptor reservoirs for direct contact with the surface of the tissue membrane.

[0139] A pre-weighed sterilised Pasteur pipette was used to gently spread the sample onto the surface of the skin ensuring it covered the entire surface. The pipette was then reweighed to obtain the weight of any residue removed from the Franz cell apparatus. The timer was then started from 0 min.

[0140] At each specified time-point (0, 30, 60, 90, 120, 150, 180, 240, and 300 min), all of the buffer phase (2 mL) was removed from the receptor chamber via a sterile syringe and needle. The sample aliquot was then flash frozen (with liquid nitrogen), and placed in the freezer (-20 °C) ready for chromatographic analysis. The receptor chamber was immediately replenished with fresh buffer solution (2 mL, equilibrated to 37 °C), ensuring complete filling of the chamber with no air bubbles.

[0141] After the final sampling time point (five hours), the stirrer was stopped and the Franz cell apparatus was carefully dismantled. The removed skin tissue sample was placed into a clean dry beaker and the donor chamber into a separate beaker.

[0142] Buffer solution (10 mL) was added to each beaker to thoroughly rinse the surfaces of the skin and donor chamber. The washings were then each flash frozen with liquid nitrogen and placed in the freezer (-20 °C) ready for chromatographic analysis.

[0143] The frozen skin samples were freeze dried, milled to fine powder, and weighed ready for chromatographic analysis. GC / MS protocol

[0144] Agilent 7890A Gas Chromatography coupled with Agilent 5975C MDS mass spectrometer and GC column DB-5ms (30.0 m x 250 pm x 0.25 pm nominal) Ultra inert size with constant follow of Helium gas (1 mL / min) in split mode (10:1) was utilised for all experiments.

[0145] All time point aliquot samples were allowed to come to room temperature and the CDB extracted with chloroform (2.0 mL) and dried with magnesium sulphate (ca. 0.2 g). The aqueous layer was discarded using an aspiration station (Gilson, UK) and the organic layer was transferred (1 mL) to amber GC / MS sample vials ready for analysis.

[0146] The samples (10 mL) collected from the washed Franz cell apparatus were extracted into chloroform (4.0 mL) and dried with magnesium sulphate (ca.

[0147] 0.4 g). The aqueous layer was discarded using an aspiration station (Gilson, UK) and the organic layer was transferred (1 mL) to amber GC / MS sample vials ready for analysis.

[0148] Samples (10 mL) collected from the washed skin were extracted into chloroform (2.0 mL), dried with magnesium sulphate (ca. 0.2 g) and filtered before being transferred (1 mL) to amber GC / MS sample vials ready for analysis.

[0149] The powdered freeze dried skin samples were dispersed into chloroform (3x 1.0 mL), filtered, dried with magnesium sulphate (ca. 0.2 g), and transferred (1 mL) to amber GC / MS sample vials ready for analysis.

[0150] The samples were loaded into the autosampler of the GM / MS. The GM / MS instrument was programmed to heat (300 °C with ramp 10 °C / min) from initial temperature (130 °C) and held for 2 mins. The total run time per sample was 24 mins. The mass spectrometer was operated in full scan mode with scanning range of 40-450 amu and an ionisation energy of 70 eV.

[0151] Results

[0152] Franz cell analysis studies of CDB permeability across porcine mucosal Tissue.

[0153] 1. CBD permeation study over time

[0154] Table 5. The total of CBD permeation across the porcine mucosal tissue membrane after 5 hours from formulations BCMS_032_020, BCMS_032_021, BCMS_032_022, BCMS_032_023, and BCMS_032_024. (EtOH - ethanol, PG - propylene glycol, MOT - medium chain triglycerides.)

[0155]

[0156] Table 6. The accumulative concentration of CBD permeation across the

[0157] porcine mucosal tissue membrane over time from formulations BCMS_032_020, BCMS_032_021, BCMS_032_022, BCMS_032_023, and BCMS_032_024.

[0158]

[0159] Referring to Figure 5, Franz cell analysis of CBD permeation studies across ex vivo porcine mucosa tissue membrane demonstrates that one embodiment of the cannabinoid-based formulation of the invention, sample BCMS_032_020, which contains 10% CBD, 0.250% terpenes, and 100 % MCT, shows a higher percentage of CBD penetration across the porcine mucosal membrane, followed by another embodiment of the cannabinoid-based formulation of the invention, sample BCMS_032_021, which contains 10% CBD, 0.250% terpenes and 100 % MCT. This confirms that the presence of terpenes increases the rate of CBD penetration across the skin membrane. The CBD permeation from the formulations based on Ethanol: Propylene glycol (50:50), namely samples BCMS_032_022 and BCMS_032_023, are significantly lower compared to the MCT formulations. Moreover, the sample BCMS_032_024 which consists of EtOH: Propylene glycol (50:50) along with terpenes, also demonstrated a reduction in CBD penetration compared to the MCT formulations.

[0160] 5 Example 8 - MRX1 formulation substantially lacks THC

[0161] Analysis of an embodiment of the cannabinoid-based formulation of the invention was performed by BCM Analytical Services, who are the contract testing business of the Fareva group, and provide full spectrum pharmaceutical analysis and testing.

[0162] Using HPLC, BCM Analytical Services tested an embodiment of the cannabinoid- 0 based formulation of the invention for impurities including d9-THC and d8-THC.

[0163] As shown in Table 7 below, the results of HPLC analysis of the formulation showed that both d9-THC and d8-THC were below the limit of detection (<LOD), thus confirming that the formulation of the invention substantially lacks THC.

[0164] 5

[0165] Table 7. HPLC impurity profile analysis of an embodiment of the cannabinoid-based formulation of the invention.

[0166] < < <

[0167]

[0168] <

[0169] <

[0170] 0 Example 9 - Treatment of Chemotherapy Induced Peripheral Neuropathy (CIPN)

[0171] An embodiment of the cannabinoid-based formulation of the invention, which may be referred to herein as MRX1, will be used to treat Chemotherapy Induced Peripheral Neuropathy (CIPN) in an upcoming Phase II placebo controlled doubleblind cross-over trial. The Phase II clinical trial will begin in 2024 and will be

[0172] 5 conducted by the University of Edinburgh. It will be led by Professor Marie Fallon, the St Columba's Hospice Chair of Palliative Medicine and Honorary Consultant in Palliative Care at the Western General Hospital in Edinburgh, Scotland. Professor Fallon is a world-leading research, and has conducted previous clinical trials using

[0173] CBD.

[0174] 0

[0175] The Applicant of the present application has signed a Drug Supply Agreement with the University of Edinburgh and the Lothian Health Board on 7 November 2023. This Phase II trial will assess the efficacy of MRX1 in providing effective analgesia in 92 patients with CIPN, as well as a range of secondary outcomes including motor function, quality of life, anxiety and depression, and sleep quantity and efficiency. The design of the trial is shown in Figure 6.

[0176] The trial will allow the inventors to demonstrate that the cannabinoid-based formulation of the invention, including embodiments such as MRX1, is efficacious in treating CIPN over a period of 5 weeks, as well as improving a range of secondary outcomes.

[0177] MRX1 will be administered using an appropriate dosing regimen and titration schedule based on available non-clinical and clinical evidence, as well as data gathered from the Phase I pharmacokinetic study that will be undertaken on MRX1 in 2024, which is discussed further in Example 12.

[0178] Data will be gathered on inflammatory markers, to investigate mechanism of action of CBD in the prevention and treatment of CIPN, and dose-response pharmacodynamics using specific and potentially novel biomarkers for CIPN disease state severity, to assist in the evaluation of efficacy and effective dose.

[0179] Brain connectome and central nervous system inflammation by fMRI will also be investigated, providing specific and potentially novel biomarkers for CIPN disease state and severity to assist in the evaluation of efficacy and effective dose.

[0180] Following the completion of this Phase II trial, the inventors intend to progress MRX1 to a further Phase III trial to demonstrate the efficacy of MRX1 in treating CIPN in a larger population. The inventors also intend to engage with the Medicines and Healthcare Products Regulatory Agency (MHRA) and National Institute for Health and Care Excellence (NICE) to progress the consideration of MRX1 for marketing authorisation to treat CIPN, NICE appraisal, and sale as a licensed drug in the United Kingdom through the NHS.

[0181] Example 10 - Treatment of pain associated with endometriosis

[0182] An embodiment of the cannabinoid-based formulation of the invention, which may be referred to herein as MRX1, will be used to treat the pain associated with endometriosis in an upcoming Phase II double blind placebo-controlled trial. The Phase II clinical trial will begin in 2024, and will be conducted by the University of Edinburgh. It will be led by Dr Lucy Whitaker, Senior Clinical Research Fellow at the Centre for Reproductive Health, University of Edinburgh.

[0183] The Applicant of the present application signed a Drug Supply Agreement with the University of Edinburgh and Lothian Health Board on 22 February 2024.

[0184] This Phase II trial will assess the efficacy of MRX1 in treating the pain associated with endometriosis in 100 patients with endometriosis identified at laparoscopy or imaging (performed in the last five years) and chronic pelvic pain of greater than six months duration. The trial will investigate the feasibility of recruitment and retention. It will also assess other symptoms, including fatigue, and the impact on quality of life and number of health care visits.

[0185] The design of the trial is a 12-week, double-blind, placebo-controlled trial with no cross-over. It will be conducted across two sites: NHS Lothian and NHS Grampian.

[0186] The trial will allow the inventors to demonstrate that MRX1 is efficacious in treating endometriosis associated pain over a period of 12 weeks, as well as improving a range of secondary outcomes outlined above. It will also record patient use of opioids before and during the treatment period to gather data on the ability of MRX1 to reduce opioid use.

[0187] MRX1 will be administered using an appropriate dosing regimen and titration schedule based on available non-clinical and clinical evidence, as well as data gathered from a Phase I pharmacokinetic study that will be undertaken on MRX1 in 2024, which is discussed further in Example 12.

[0188] Blood samples will be taken for testing of inflammatory markers to investigate mechanism of action of CBD in the prevention and treatment of pain associated with endometriosis.

[0189] Following the completion of this Phase II trial, the inventors intend to progress MRX1 to a further Phase III trial to demonstrate the efficacy of MRX1 in treating pain associated with endometriosis in a larger population. They also intend to engage with the Medicines and Healthcare Products Regulatory Agency (MHRA) and National Institute for Health and Care Excellence (NICE) to progress the consideration of MRX1 for marketing authorisation to treat endometriosis, NICE appraisal, and sale as a licensed drug in the United Kingdom through the NHS.

[0190] Example 11 - Resolution of pathogenic nature of fibroblasts and mesenchymal stromal cells in endometriosis

[0191] Fibroblasts play a significant role in endometriosis, contributing to its complex pathogenesis. Briefly, fibroblasts affect the remodelling of the extracellular matrix (ECM), contribute to inflammatory processes, modulate angiogenesis, and are known to be immunomodulatory. All these processes are involved in endometriosis, and there is evidence of fibroblast involvement in all these processes.

[0192] Utilising an embodiment of the cannabinoid-based formulation of the invention, which may be referred to herein as MRX1, a co-funded PhD studentship at the University of Reading under the supervision of Professor Darius Widera will perform, and investigate, the following:

[0193] 1) Inflammation will be simulated by exposing fibroblasts / mesenchymal stem / stromal cells (MSCs) to Tumour Necrosis Factor-o, IL-ip, and IL-6, either alone, or in combination with MRXlxfollowed by immunocytochemical staining against the NF-KB subunit p65. Imaging and analysis of NF-KB translocation will be performed on a Revvity Operetta high-content imaging system (with machine learning capacity).

[0194] 2) Secretion of pro-angiogenic and pro-inflammatory factors will be assessed by ELISA (e.g., FGF-2, IL-6, IL-lg, VEGF).

[0195] 3) Secretomes from fibroblasts / MSCs treated as described in 2) will be collected and used in a HUVEC in vitro angiogenesis model.

[0196] 4) Secretomes generated as above will be used in a human monocyte polarisation assay.

[0197] 5) Fibroblast and MSCs will be treated as described in 1), and the composition of the deposited ECM (Collagens, Proteoglycans, and Glycoproteins (e.g., fibronectin, laminin), Matrix Metalloproteinases (MMPs), Integrins, and Basement Membrane Components) will be studied using immunocytochemistry and high-content imaging (see 1 above).

[0198] The inventors hypothesise that MRX1 can reduce inflammatory signalling, production of pro-angiogenic factors, and ECM remodelling by fibroblasts and MSCs, and modulate immune cells towards a regulatory M2-phenotype. Example 12 - Phase I pharmacokinetic (PK) study

[0199] An embodiment of the cannabinoid-based formulation of the invention, which may be referred to herein as MRX1, will be administered to healthy volunteers to determine the safety and PK profile of the MRX1 formulation. MRX1 will be administered to between 20 and 30 participants using a dosing regimen and titration schedule aligned with non-clinical and clinical evidence, and the inventors' expected administration of the MRX1 formulation in clinical practice to treat various chronic inflammatory pain conditions.

[0200] The Phase I PK study will be conducted in 2025 by a phase 1 clinical research unitin Australia. MRX1 will be administered to participants over a period of 1-3 weeks. Blood samples will be taken at regular intervals, and those blood samples will be tested for analytes including: Cannabidiol (CBD), 7-Carboxy cannabidiol (7-COOH-CBD), 7-Hydroxy cannabidiol (7-OH-CBD), Delta-9 Tetrahydrocannabinol (THC), ll-carboxy-A9-THC (11-OH-THC), and 11-hydroxy- A9-THC (11-COOH-THC).

[0201] In line with the novel and innovative characteristics of the cannabinoid-based formulation of the invention, and the fact that the formulation substantially lacks THC, the inventors expect that there will be a low or zero level of Delta-9 Tetrahydrocannabinol (THC) and metabolites detected in the blood samples of participants. This would provide further evidence of the novelty of the cannabinoid-based formulation of the invention when administered to humans.

[0202] Based on their research and the results of the permeability study previously conducted on the cannabinoid-based formulation of the invention, as discussed in Example 7, the inventors also expect that the cannabinoid-based formulation of the invention, such as MRX1, will deliver superior CBD bioavailability to that of other CBD in oil formulations. The presence of the proprietary terpene blend of the cannabinoid-based formulation of the invention is expected to increase permeability, and therefore bioavailability, of CBD in the blood.

[0203] Following the completion of this Phase I trial, the inventors will be able to utilise these proven novel characteristics, and the bioavailability data which is generated, to better target the low dosing regimen and titration schedule used for the cannabinoid-based formulation of the invention in additional clinical trials, including two Phase II clinical trials being undertaken by the University of Edinburgh using the cannabinoid-based formulation of the invention to treat the pain associated with chemotherapy induced peripheral neuropathy and endometriosis, respectively, as discussed in Examples 10 and 11.

[0204] Example 13 - Low CBD dose treatment of pain

[0205] Two chemotherapy induced peripheral neuropathy (CIPN) patients experienced significant improvements in pain, mood, sleep and general quality of life when administered a low dose of CBD. The preparation administered to these patients was a lOOmg / ml CBD oil formulation.

[0206] Patient 1

[0207] Patient 1 was a 53-year-old woman weighing 75kg. The patient had a 3-year history of intractable, severe chemotherapy induced peripheral neuropathy and was 3-years post treatment for breast cancer with curative intent. Pain management to date had included Gabapentin 600mg ter in die (t.i.d), Tramadol 50mg t.i.d, and Paracetamol 1g q.i.d. The patient had also had physiotherapy and topical treatments such as topical menthol 5% cream, topical lidocaine 5% patches and topical capsaicin. The patient had not experienced any improvement in scores on these drugs.

[0208] The patient consistently took a dose of 60mg bd CBD and in less than 4 weeks had the following improvements:

[0209]

[0210] *A reduction in score denotes an improvement i.e. 8 / 10 quality of life is a poorer quality of life than 3 / 10.

[0211] Patient 2

[0212] Patient 2 was a 45-year-old woman weighing 68kg. The patient had a 2-year history of severe chemotherapy induced peripheral neuropathy following treatment for breast cancer with curative intent. Several neuropathic agents had been tried and failed with this patient including pregabalin, duloxetine, amitriptyline, paracetamol, NSAIDs and weak opioid, codeine. The patient had also tried topical menthol 5%, topical lidocaine patches 5% and physiotherapy. The patient had not experienced any improvement in scores on these drugs and had stopped all treatments.

[0213] The patient consistently took a dose of 70mg bd CBD and in less than 4 weeks had the following improvements:

[0214]

[0215] *A reduction in score denotes an improvement i.e. 8 / 10 quality of life is a poorer quality of life than 1 / 10.

[0216] The patient had no side effects.

[0217] In view of the above data, the inventors recommend a dose of 0.5mg CBD / kg body weight / day - 12.5mg CBD / kg body weight / day, which is ideally split into 2 doses per day, i.e. 0.25mg CBD / kg bw b.i.d. - 6.25mg CBD / kg bw b.i.d.

[0218] Typically, the dose is: 1.9mg CBD / kg bw / day, ideally split into 2 doses per day, i.e.

[0219] 0.95mg / kg bw b.i.d.

[0220] Conclusions

[0221] The resultant CBD formulations are referred to as Cannabis-Based Products for Medicinal use (CBPM) in humans or animals. In particular, the CBPMs comprises CBD isolate, and optionally terpenes and medium-chain triglyceride (MCT) or sesame oil in a liquid form for oral delivery, and is administered at low dose, such as 60-70mg CBD / day, or 0.8mg / kg bd CBD-lmg / kg bd CBD. The use of these formulations at such low doses focuses primarily on conditions associated with pain (in particular, CIPN), inflammation, autoimmune diseases, neurological diseases, neoplasm, nausea, hypertension, hypertrophic cardiomyopathy, and fibrosis. The formulations (or CBPMs) are broad-spectrum CBD mixtures, which consist of a particular combination of CBD and terpenes, produced at pharmaceutical standards under the guidelines of Good Manufacturing Practice (GMP) and designed to produce precise physical effects. To date, there are no products available in the UK that comply with the GMP standard and which can be used at such low doses, and so the pharmaceutical formulations of the invention address this gap and facilitate the access and delivery of a pharmaceutical standard CBPM for use in clinical trials and for treating patients.

[0222] The preparation of these liquid formulations for sublabial, buccal or sublingual delivery allows the introduction of the drug into the systemic circulation via a mucous membrane, increasing the onset of activity and potency due to bypassing first-pass metabolism and increasing the drug bioavailability. Advantageously, given the lipophilicity and poor bioavailability of oral administration of cannabinoids for gastrointestinal absorption, which is considered the most popular method currently in the market for the delivery of medicines, the formulations of the invention surprisingly enable administration by this route.

Claims

1. Claims1. A composition comprising cannabidiol (CBD), for use in treating, preventing or alleviating pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea in a subject, wherein a dosage of between O.lmg and 20mg CBD per kilogram subject body weight per day is administered to the subject.

2. A composition, for use according to claim 1, wherein:4.(a) between O.lmg and 18mg CBD per kilogram subject body weight per day is administered to the subject;5.(b) between O.lmg and 16mg CBD per kilogram subject body weight per day is administered to the subject;6.(c) between O.lmg and 14mg CBD per kilogram subject body weight per day is administered to the subject;7.(d) between O.lmg and 12mg CBD per kilogram subject body weight per day is administered to the subject.8.(e) between O.lmg and llmg CBD per kilogram subject body weight per day is administered to the subject; or9.(f) between O.lmg and lOmg CBD per kilogram subject body weight per day is administered to the subject;3. A composition, for use according to any preceding claim, wherein:11.(a) between O.lmg and 9mg CBD per kilogram subject body weight per day is administered to the subject;12.(b) between O.lmg and 8mg CBD per kilogram subject body weight per day is administered to the subject;13.(c) between O.lmg and 7mg CBD per kilogram subject body weight per day is administered to the subject,14.(d) between O.lmg and 6mg CBD per kilogram subject body weight per day is administered to the subject;15.(e) between O.lmg and 5mg CBD per kilogram subject body weight per day is administered to the subject:16.(f) between O.lmg and 4mg CBD per kilogram subject body weight per day is administered to the subject.

4. A composition, for use according to any preceding claim, wherein:(a) between O.lmg and 3mg CBD per kilogram subject body weight per day is administered to the subject;18.(b) between O.lmg and 2mg CBD per kilogram subject body weight per day is administered to the subject; or19.(c) between O.lmg and Img CBD per kilogram subject body weight per day is administered to the subject.

5. A composition, for use according to any preceding claim, wherein:21.(a) between 0.15mg and 4.5mg CBD per kilogram subject body weight per day is administered to the subject;22.(b) between 0.2mg and 4mg CBD per kilogram subject body weight per day is administered to the subject;23.(c) between 0.3mg and 3.5mg CBD per kilogram subject body weight per day is administered to the subject; or24.(d) between 0.4mg and 3mg CBD per kilogram subject body weight per day is administered to the subject.

6. A composition, for use according to any preceding claim, wherein:26.(a) between 0.45mg and 2.5mg CBD per kilogram subject body weight per day is administered to the subject;27.(b) between 0.50mg and 2mg CBD per kilogram subject body weight per day is administered to the subject;28.(c) between 0.55mg and 1.75mg CBD per kilogram subject body weight per day is administered to the subject; or29.(d) between 0.60mg and 1.5mg CBD per kilogram subject body weight per day is administered to the subject.

7. A composition, for use according to any preceding claim, wherein:31.(a) between 0.65mg and 1.3mg CBD per kilogram subject body weight per day is administered to the subject;32.(b) between 0.7mg and l.lmg CBD per kilogram subject body weight per day is administered to the subject; or33.(c) between 0.8mg and Img CBD per kilogram subject body weight per day is administered to the subject.

8. A composition, for use according to any preceding claim, wherein:(a) about 0.8mg CBD per kilogram subject body weight per day is administered to the subject or35.(b) about Img CBD per kilogram subject body weight per day is administered to the subject.

9. A composition, for use according to any preceding claim, the composition is administered to the subject as a single dose per day.

10. A composition, for use according to any preceding claim, wherein the composition is administered to the subject as two or more doses per day.

11. A composition, for use according to any preceding claim, wherein:39.(a) 0.4mg CBD / kg body weight bis in die (b.i.d.) is administered; or40.(b) 0.5mg CBD / kg body weight bis in die (b.i.d.) is administered.

12. A composition, for use according to any preceding claim, wherein:42.(a) between 0.2mg and lOmg CBD / kg body weight bis in die (b.i.d.) is administered; or43.(b) between 0.4mg and 5mg CBD / kg body weight bis in die (b.i.d.) is administered.

13. A composition, for use according to any preceding claim, wherein the dose is 1.9mg CBD / kg bw / day, optionally split into two doses per day, i.e. 0.95mg / kg bw b.i.d.

14. A composition, for use according to any preceding claim, wherein:46.(a) between 30 and lOOmg CBD per day is administered to the subject;47.(b) between 40 and 90mg CBD per day is administered to the subject; or48.(c) between 50 and 80mg CBD per day is administered to the subject.

15. A composition, for use according to any preceding claim, wherein:50.(a) between 55 and 75mg CBD per day is administered to the subject; or51.(b) between 60 and 70mg CBD per day is administered to the subject.

16. A composition, for use according to any preceding claim, wherein the pain, inflammation, an autoimmune disease, a neurological disease, neoplasm, or nausea are selected from the group of conditions consisting of: chemotherapy-induced peripheral neuropathy (CIPN), endometriosis, osteoarthritis, rheumatoid arthritis,ulcerative colitis, Crohn's disease, fibromyalgia, irritable bowel syndrome, asthma, chronic obstructive pulmonary disease (COPD), gout, scleroderma, lupus, Ehlers-Danlos syndrome, pericarditis, myocarditis, myocardial infarction, chemotherapy-induced nausea, anxiety, Type I diabetes, cardiovascular disease, primary hypertension, secondary hypertension, resistant hypertension, isolated systolic hypertension, malignant hypertension, obstructive hypertrophic cardiomyopathy, nonobstructive hypertrophic cardiomyopathy, pulmonary fibrosis, liver fibrosis, heart fibrosis, kidney fibrosis, mediastinal fibrosis, retroperitoneal cavity fibrosis, bone marrow fibrosis, skin fibrosis, and scleroderma.

17. A composition, for use according to any preceding claim, wherein the composition is used to treat, prevent or alleviate pain.

18. A composition, for use according to any preceding claim, wherein the composition is used to treat, prevent or alleviate chemotherapy-induced peripheral neuropathy (CIPN).

19. A composition, for use according to any preceding claim, wherein the formulation is administrable by oral, sublabial, buccal, sublingual, or oropharyngeal administration delivery.

20. A composition, for use according to any preceding claim, wherein the formulation substantially lacks any THC.

21. A composition, for use according to any preceding claim, wherein the concentration of THC in the composition is less than lOOOpg per finished product.

22. A composition, for use according to any preceding claim, wherein the concentration of THC in the composition is less than 0.01% (w / v), less than 0.005% (w / v), or less than 0.001% (w / v).

23. A composition, for use according to any preceding claim, wherein the concentration of THC in the composition is less than 0.0005% (w / v), less than 0.0002% (w / v), or less than 0.0001% (w / v).

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