Cannabidiol compositions and uses thereof

Abstract

The present application relates to cannabidiol composition and its use. Specifically, the present application relates to cannabidiol composition containing cannabidiol and surfactant, which can significantly improve the water solubility of cannabidiol, making it better applied in the fields of medicine, daily chemical, food, health care products, etc.

Description

Technical Field

[0001] This application relates to the field of chemistry, and in particular to a composition containing cannabidiol (CBD) that can significantly improve the water solubility of cannabidiol, making it better suited for use in the pharmaceutical field. Background Technology

[0002] Cannabis (scientific name: Cannabis sativa L.) is a plant belonging to the Cannabaceae family and the Cannabis genus. Also known as hemp, Chinese hemp, fire hemp, mountain silk hemp, and yellow hemp, it has important agricultural and medicinal value. Cannabis contains a toxic component called tetrahydrocannabinol (THC), which can cause hallucinations and addiction, and can be used as a drug. Its cultivation was prohibited for a considerable period of time.

[0003] Hemp varieties intended for industrial use (referred to as "industrial hemp") contain less than 0.3% THC in their flowers and leaves during their growth period. They are not valuable for extracting the toxic component THCTHC or for direct use as drugs. However, given their extremely high economic and medicinal value, they can be legally cultivated and industrially developed.

[0004] Currently, over 500 substances have been isolated from cannabis plants, including at least 86 cannabinoids. Cannabinoids are a class of substances unique to cannabis plants and are the main active ingredients; research on them has always been a hot topic in cannabis research. The main cannabinoids in cannabis plants include THC, cannabinol (CBN), CBD, cannabinoid glycosides (CBG), and cannabinoid cyclic glycosides (CBC), with the first three accounting for over 90% of cannabinoids.

[0005] Cannabidiol is a type of cannabinoid with the molecular formula C6H2O. 21 H 30 O2 is a pale yellow resin or crystal, usually extracted from the natural plant cannabis, but can also be synthesized artificially. It is a compound with diverse pharmacological activities, almost insoluble in water, but soluble in organic solvents such as ethanol, methanol, ether, benzene, chloroform, and petroleum ether. Its structural formula is shown in Formula I below:

[0006]

[0007] Cannabidiol is completely different from tetrahydrocannabinol. Not only does it have no hallucinogenic effects, but it also has good pharmacological activity in treating spasms, anxiety / depression, inflammation, cancer, rheumatoid arthritis, multiple sclerosis, epilepsy, especially refractory epilepsy, as well as antiviral and analgesic effects. Therefore, it has become a hot topic of research in the global medical field.

[0008] However, cannabidiol suffers from poor water solubility, low bioavailability, and weak stability, which greatly limits its application, as follows:

[0009] GW Pharma is a company primarily engaged in the research and development of cannabis-related drugs. Its proprietary combination drug Sativex, containing cannabidiol (CBD) and THC, has been developed and marketed for the treatment of multiple sclerosis. However, due to the poor stability and low bioavailability of CBD when administered via the gastrointestinal system, it has been developed as an oral spray for absorption through the oral mucosa. Furthermore, due to the water solubility of CBD, the company has had to use organic solvents such as anhydrous ethanol and propylene glycol for formulation.

[0010] However, according to information disclosed by Sativex in its Summary of Product Characteristics, despite choosing oral mucosal administration and an organic solvent system, Sativex still has the following issues:

[0011] 1) Pharmacokinetic behavior is greatly affected by whether or not food has been consumed. Compared with fasting conditions, the C60 of cannabidiol is significantly higher in the cannabidiol than in fasting conditions. max The difference between AUC and AUC was 3.3 times and 5.1 times, respectively;

[0012] 2) It exhibits significant differences in pharmacokinetic characteristics among different subjects, with a large coefficient of variation. The C60 of cannabidiol after a single dose... max The CV% was 64.1%, and the AUC CV% was 72.5%. After multiple consecutive administrations, the C of cannabidiol... max The CV% was 75.7%, and the CV% of AUC was 46.6%.

[0013] 3) It also exhibits high variability in pharmacokinetic behavior within the same subject.

[0014] These issues are related to the properties of cannabidiol itself, and of course, also to the specific route of administration and formulation technology chosen by Sativex. In summary, these significant uncertainties lead to substantial differences in clinical efficacy and inevitably pose certain safety risks.

[0015] Russell Hobart Stebbins disclosed a method for preparing solid cannabidiol in patent US20160143972 A1, claiming that this form of cannabidiol is soluble in an aqueous system. Although the above-mentioned prior art has improved the problems of poor water solubility and low bioavailability of cannabidiol to some extent, the effect is still unsatisfactory.

[0016] Furthermore, it should be pointed out that, in addition to its poor water solubility, issues regarding the safety, compliance, cost-effectiveness, and stability of cannabidiol are also unavoidable:

[0017] 1) Cannabidiol has low bioavailability and requires high doses to achieve efficacy. However, high doses inevitably lead to compliance problems and increase the economic burden on patients. At the same time, high doses can also increase the burden on the liver and kidneys, leading to unpredictable liver and kidney diseases.

[0018] 2) Cannabidiol is highly lipid-soluble (K oil-water = 6-7), with an apparent volume of distribution of about 32 L / kg. It can be rapidly distributed to the brain, adipose tissue and other organs. Cannabidiol has a high plasma protein binding rate, and about 10% can bind to red blood cells. Long-term use may lead to accumulation in the body of patients, especially obese patients.

[0019] 3) In their article "Identification of Psychoactive Degradants of Cannabidiol in Simulated Gastric and Physiological Fluid," John Merrick, Brian Lane, and others disclosed the effects of cannabidiol in simulated gastric fluid. They pointed out that under these experimental conditions, 85% of cannabidiol was degraded in 60 minutes and 98% was degraded in 120 minutes. This is very unfavorable. Even more alarming is that after being degraded in simulated gastric fluid, cannabidiol is converted into the psychoactive substance tetrahydrocannabinol (THC), which is extremely dangerous and will inevitably bring huge safety hazards in practical applications.

[0020] In summary, finding suitable technical means to solve the problems existing in the application of cannabidiol has become an urgent problem to be solved in this field. Summary of the Invention

[0021] Therefore, on the one hand, this application provides a cannabidiol composition containing cannabidiol and a surfactant.

[0022] In some preferred embodiments, the surfactant is selected from nonionic surfactants.

[0023] In some preferred embodiments, the surfactant is selected from adducts based on polypropylene glycol and / or polyethylene oxide, as well as polyol-type esters or amide adducts.

[0024] The polypropylene glycol mentioned in this article refers to... The polymer has a molecular weight of 1000-10000 Da, preferably 5000-10000 Da, and more preferably 6000-9000 Da.

[0025] The polyoxyethylene mentioned in this article refers to the polyoxyethylene with... The polymer has a molecular weight of 500-2000 Da, preferably 1000-2000 Da.

[0026] In some preferred embodiments, the polyol-type ester adduct refers to an ester formed by organic compounds containing multiple hydroxyl groups in molecules such as ethylene glycol, sorbitol, and sucrose with higher fatty acids, such as sorbitol esters and sucrose esters.

[0027] In some preferred embodiments, the polyol-type amide adduct refers to an organic compound formed by the condensation of a polyol amine (e.g., diethanolamine) with a fatty acid, wherein the fatty acid may be coconut oil acid, fatty acid, or lauric acid.

[0028] In some preferred embodiments, the surfactant is poloxamer.

[0029] In some preferred embodiments, the poloxamer has a molecular weight of 9000–20000 Da, for example 9000–16000 Da, 9000–12600 Da, 9900–12600 Da, or 10000–15000 Da (preferably 9950–12600 Da), and the percentage of polyoxyethylene blocks is 40%–80%, for example 60%–80%, 60%–75%, 62%–72%, 65%–75%, or 65%–80%.

[0030] In some preferred embodiments, the poloxamer is P407 or a poloxamer with a molecular weight of 9950-12600 Da and a polyoxyethylene block percentage of 70%.

[0031] In some preferred embodiments, the mass ratio of cannabidiol to surfactant is 1:(1-100).

[0032] In some preferred embodiments, the mass ratio of cannabidiol to surfactant is 1:(5-50).

[0033] In some preferred embodiments, the composition also contains an antioxidant.

[0034] In some preferred embodiments, the antioxidant is an organic acid or a pharmaceutically acceptable salt thereof.

[0035] In some preferred embodiments, the antioxidant is selected from citric acid, tartaric acid, malic acid, succinic acid, ascorbic acid, and pharmaceutically acceptable salts thereof.

[0036] In some preferred embodiments, the antioxidant is selected from citric acid, tartaric acid, malic acid, succinic acid, ascorbic acid, potassium citrate, sodium citrate, and potassium hydrogen tartrate.

[0037] In some preferred embodiments, the antioxidant is citric acid or tartaric acid.

[0038] In some preferred embodiments, the weight ratio of cannabidiol to antioxidant is 1:(0.01-0.2); for example, 1:(0.01-0.1).

[0039] In some preferred embodiments, the weight ratio of cannabidiol, surfactant, and antioxidant is 1:(1-100):(0.01-0.2).

[0040] In some preferred embodiments, the weight ratio of cannabidiol, surfactant, and antioxidant is 1:(5-50):(0.01-0.1).

[0041] In some preferred embodiments, the weight ratio of cannabidiol, surfactant, and antioxidant is 1:50:0.01.

[0042] In some preferred embodiments, the composition further contains a pharmaceutically acceptable carrier or excipient.

[0043] The carriers described herein include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycerol, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, and lanolin. The excipients referred to herein are additives in pharmaceutical preparations other than the active pharmaceutical ingredient. They are stable in nature, have no incompatibilities with the active pharmaceutical ingredient, do not produce side effects, do not affect efficacy, are not easily deformed, cracked, moldy, or infested by insects at room temperature, are harmless to the human body, have no physiological effects, do not produce chemical or physical reactions with the active pharmaceutical ingredient, and do not affect the content determination of the active pharmaceutical ingredient. Examples of excipients include binders, fillers, disintegrants, and lubricants in tablets; wine, vinegar, and medicinal juice in traditional Chinese medicine pills; the base portion in semi-solid preparations such as ointments and creams; and preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, osmotic pressure regulators, and colorants in liquid preparations.

[0044] In some preferred embodiments, the composition further contains 5-20 times the weight of water as solids; preferably 10-15 times; more preferably 10 times.

[0045] The solids mentioned herein refer to the substances remaining after the composition or product has been thoroughly dried, that is, the sum of substances in the composition excluding water.

[0046] In some preferred embodiments, the composition is a nanoemulsion formulation.

[0047] In some preferred embodiments, the nanoemulsion has a particle size of 1-200 nm.

[0048] In some preferred embodiments, the nanoemulsion has a particle size of 1-100 nm.

[0049] In some preferred embodiments, the composition is a tablet, granule, capsule, soft capsule, or injection.

[0050] In some preferred embodiments, the composition can be administered orally, transdermally, or by injection.

[0051] In another aspect, this application relates to the use of the said composition in the preparation of medicaments, antiviral drugs, or analgesics for the prevention and / or treatment of spasms, anxiety / depression, inflammation, cancer, rheumatoid arthritis, multiple sclerosis, epilepsy, especially refractory epilepsy.

[0052] Unless otherwise specified, "molecular weight" as used in this article specifically refers to "number-average molecular weight Mn".

[0053] Beneficial effects of the invention

[0054] This application provides a cannabidiol composition that can achieve at least one of the following technical effects:

[0055] 1) The composition can significantly improve the water solubility of cannabidiol, which greatly promotes its application in the pharmaceutical field.

[0056] 2) The composition has high stability, solving the problem of cannabidiol's instability and easy degradation in gastric juice;

[0057] 3) The composition can significantly improve the relative bioavailability of cannabidiol;

[0058] 4) The pharmacokinetics of cannabidiol in the body are more consistent, with lower C-values ​​among different individuals. max The low coefficients of variation of the two key pharmacokinetic indicators, AUC, reduce the unpredictable risks in clinical application.

[0059] 5) The binding rate of cannabidiol to plasma proteins in the composition is significantly lower than that of cannabidiol alone, which solves the problem of accumulation of cannabidiol with long-term use, which has been reported in the media. Attached Figure Description

[0060] Figure 1 The GPC chromatogram of poloxamer P-HY prepared in this application is shown. Detailed Implementation

[0061] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0062] Cannabidiol: Yunnan Hansu Biotechnology Co., Ltd.

[0063] Example 1: Preparation and performance parameter determination of self-made poloxamer P-HY

[0064] 1. Synthetic route of poloxamer (P-HY):

[0065] 1)

[0066] 2)

[0067] (1) Condensate propylene oxide onto 1,2-propanediol to obtain polyoxypropylene intercalation segment; wherein the alkaline 1,2-propanediol is used as an initiator, and the molar ratio of each substance is propylene oxide: 1,2-propanediol: KOH / NaOH = (220-320): 1: (2-0.5), the reaction temperature is 90-120℃, and the reaction endpoint (time) varies depending on different reaction conditions, with the reference standard being the end of the reflux state.

[0068] (2) Ethylene oxide is condensed to both ends of the polyoxypropylene intercalation segment obtained in step (1) under the catalysis of potassium hydroxide (or sodium hydroxide) and EDTA. The molar ratio of the starting materials is controlled as ethylene oxide: polyoxypropylene: potassium hydroxide (or sodium hydroxide): EDTA = (450-650): 1: (0.5-1): (0.05-0.1). The reaction temperature is 90-120℃ and the time is 9-20h. Finally, poloxamer (named P-HY) with a molecular weight range of 9954-12506 Da and a polyoxyethylene segment of 62-72% is obtained. The final total yield is 45%-50%.

[0069] 2. Specific preparation method of poloxamer P-HY

[0070] (1) Preparation of polyoxypropylene intercalation segment

[0071] The reaction was carried out according to the material ratio in Table 1. First, anhydrous sodium sulfate dehydrated propylene glycol was added sequentially to the reactor as an initiator, along with KOH / NaOH fine powder (80-100 mesh) dried under reduced pressure. Nitrogen was used to purge the air, stirring was started, and hot water (95-100℃) was introduced for heating. When the KOH / NaOH was completely dissolved, propylene oxide was slowly added, and the reaction began. When the reflux phenomenon in the reactor ended, the hot water supply was stopped, and tap water was circulated to cool the reactor, resulting in a yellow, viscous, crude polyoxypropylene with a molecular weight of about 3500 Da. This crude product was then extracted with an equal volume of water and twice the volume of n-heptane, repeatedly washed and purified with purified water, and finally concentrated under reduced pressure to obtain a yellow, oily polyoxypropylene intercalation segment.

[0072] Table 1. Feed and Output of Polyoxypropylene Intercalation Segments

[0073]

[0074]

[0075] (2) Preparation of poloxamer P-HY

[0076] Using the polyoxypropylene intercalation segment obtained in step (1) as raw material, dry inorganic alkali powder and EDTA were added according to the feeding amounts in Table 2. Nitrogen was purged to remove air, stirring was started, and ethylene oxide was slowly added from the bottom of the reactor under boiling water bath heating. The reaction continued, and the reaction time was as described in Table 2. When the reaction was stopped, a pale yellow paste-like crude poloxamer copolymer was obtained. It was dissolved in purified water, and 1 mol / L HCl was added dropwise to adjust the pH to 6.5-7.0. The mixture was filtered through 300-500 mesh neutral alumina to remove macromolecular polymers. The filtrate was loaded onto a macroporous resin D101 column and flushed with 2 column volumes of purified water to remove salts and oligomers. Then, 3 column volumes of 15% ethanol were added to obtain the target product segment eluent. The target product segment eluent was concentrated under reduced pressure at 60°C to a thick paste, poured out, and dried under vacuum at 40°C and -0.10 MPa for 48 hours to obtain poloxamer P-HY product with a moisture content of less than 0.2%.

[0077] Table 2. Feed and output of poloxamer PH-Y preparation

[0078]

[0079] 3. Comparison of performance parameters between Poloxamer P-HY and other commercially available Poloxamer models

[0080] (1) Determination of the molecular weight of poloxamer by GPC method

[0081] Prepare a 5% solution of each poloxamer sample and determine it using a Waters 1515GPC gel permeation chromatography system. The column is a G5000PWXL column (10μm, 7.8*300mm), with tetrahydrofuran as the mobile phase, 25℃, flow rate 1ml / min, and detection at a wavelength of 256nm (detector model: Waters 2489).

[0082] The calibration curve obtained using polyoxypropylene with a known molecular weight (molecular weight: 3890 Da) as a standard is as follows:

[0083] Log M sam =9.03-0.19Vsam(R) 2 =0.998).

[0084] M sam Vsam represents the molecular weight of the sample; Vsam represents the volume of the sample elution solution.

[0085] For detailed GPC spectral data of the self-made poloxamer P-HY, please refer to [link to relevant data]. Figure 1 The P-HY segment of poloxamer is the 6th peak, with a retention time of 24.948 min. It is the main component peak, with a percentage area of ​​88.73% and an average molecular weight of 11432 Da. The molecular weight of oligomers is mainly below 8000 Da, while the molecular weight of larger polymer molecules is above 15000 Da.

[0086] The molecular weight determination results of commercially available poloxamer samples are shown in Tables 3 and 4.

[0087] (2) Determination of the content of poloxamer polyoxyethylene intercalation segment:

[0088] Take each poloxamer sample and dissolve it in deuterated methanol (or heavy water) containing 1% tetramethylsilane to prepare a 10%-20% (g / ml) solution. Take 0.5-1.0 ml of the above solution and place it in an NMR tube. Add 1 drop of heavy water, shake, and analyze the NMR spectrometer from 0*10⁻¹⁰. -6 Up to 5*10 -6 Scanning was performed, and the PEO value was calculated using the direct comparison method. The proportion of polyoxyethylene in the total molecular composition was obtained by the following formula. The results are shown in Tables 3 and 4:

[0089] PEO%=3300a / (33a+58)

[0090] In the formula, a = (A2 / A1) - 1

[0091] A1 is 1.15 * 10 -6 The integral area of ​​the bimodal peaks represents the methyl groups of polyoxypropylene;

[0092] A2 is (3.2-3.8)*10 -6The integral area of ​​the composite peak represents CH2O of polyoxypropylene, CH2O of polyoxyethylene, and CH2O of polyoxypropylene.

[0093] (3) Calculation of hydrophilic-lipophilic balance (HLB value):

[0094] The HLB of a surfactant is represented by the difference between the sum of the HLB values ​​of the individual groups of the hydrophilic and lipophilic groups.

[0095] HLB = Σ(hydrophilic group) - Σ(lipophilic group) + 7

[0096] The HLB value of oxyvinyl group is 0.33, and the HLB value of oxypropylene group is 0.15. The approximate HLB value of poloxamer was calculated using the formula, and the results are shown in Tables 3 and 4.

[0097] (4) pH value determination: Each poloxamer sample was prepared into a 2% (w / w) dilute solution, and three parallel groups were formed. The pH value was determined by a precision pH meter (model: Mettler S220), and the average value was taken. The results are shown in Table 3.

[0098] (5) Determination of the solubility of cannabidiol samples with different formulations: Each poloxamer sample to be tested was mixed with the self-made poloxamer P-HY and cannabidiol at a weight ratio of 25:1 to prepare a semi-finished product. The maximum solubility in 100 ml of purified water at room temperature (20-25℃) was determined. The mixture was shaken on a shaker to aid dissolution. The endpoint was determined when the precipitate did not dissolve within 6-8 hours. The results are shown in Table 3.

[0099] Table 3. Performance Comparison of Poloxamer P-HY Finished Products

[0100]

[0101] Table 4 Comparison of Poloxamer Specifications and Physicochemical Properties

[0102]

[0103] Example 2: Preparation and performance determination of cannabidiol composition

[0104] 1. Preparation process of cannabidiol composition

[0105] The cannabidiol composition described in this application can be prepared using the following processes:

[0106] a) Physical grinding: Weigh the prescribed amount of each component, place them in a mortar of suitable size or a grinding machine, and grind them thoroughly until the components in the prescription are evenly mixed. Then, pulverize the evenly mixed material by physical or air jet pulverization or by passing it through a pharmacopoeia sieve.

[0107] b) Heating and melting: Place each component of the prescription in a container of suitable size, heat to above 65°C to make each component of the prescription melt, then stir thoroughly at this temperature, and then cool to room temperature, and pulverize the homogeneous mixture using the method described in a).

[0108] c) Spray drying: Weigh the prescribed amounts of each component, disperse them in purified water at a solid content of 1-5%, and after uniform dispersion, spray dry them in a spray dryer to obtain powdered material. Then, pulverize the spray-dried material using the method described in a).

[0109] d) Dry granulation: Place each component of the prescription in a dry granulator for dry granulation, and finally pulverize the resulting granules using the method described in a).

[0110] e) Wet granulation: Place each component of the prescribed amount in a wet granulator, add ethanol as a wetting agent, perform wet granulation, then dry the obtained granules below 50°C to remove the ethanol, and finally pulverize the obtained granules using the method described in a).

[0111] 2. Screening of excipients: Cannabidiol compositions were prepared according to method a) with a weight ratio of poloxamer to cannabidiol of 50:1. The composition was evaluated comprehensively based on its maximum solubility, solution state, and colloidal particle size.

[0112] The cannabidiol composition, when dissolved in approximately 10 times its weight (w / w) or more of water, can form a thermodynamically stable nanoemulsion through self-emulsification. Its maximum solubility, solution state, and colloidal particle size were determined using the following methods, and the results are shown in Table 5.

[0113] 1) Maximum solubility determination method: Due to the unique solubility characteristics of poloxamer (e.g., it dissolves more readily at low temperatures, and the resulting composition has a sustained-release-like effect), the method for determining its maximum solubility differs from the standard pharmacopoeia requirements. At room temperature (20-25℃), the composition is gradually added to 100 ml of purified water in increments of 0.05 g (initially quickly then slowly), with shaking to aid dissolution. The endpoint is determined when the precipitate remains undissolved overnight (6-8 hours).

[0114] 2) State of the solution: Take each group of prescription samples and prepare a 2% (w / w) solution, and observe the physical state of the solution.

[0115] 3) Micelle particle size determination: The Mastersizer 2000 laser particle size analyzer (MS2000, 0.02-2000μm, 0.2% opacity) was used for wet measurement. The sample was prepared as a 0.1mg / ml solution and filtered through a 0.45μm filter membrane. The initial filtrate was discarded and the subsequent filtrate was the sample to be tested.

[0116] Table 5 Results of the screening of auxiliary materials

[0117]

[0118] As shown in Table 5, the solubility, solution color, and nanoemulsion particle size (<100) of the self-made P-HY (corresponding molecular weight 9950-12500 Da, polyoxyethylene segment ratio 62-72%) group of cannabidiol compositions were significantly better than those of other groups.

[0119] 3. Screening of excipient ratios: Poloxamer P-HY and P407 were preferred. Cannabidiol compositions were prepared according to method a) in 1) with poloxamer:cannabidiol weight ratios of 100:1, 50:1, 10:1, 5:1 and 1:1 respectively. The maximum solubility, solution state and colloidal particle size were used as indicators for comprehensive evaluation. The results are shown in Table 6.

[0120] Table 6 Screening Results of Auxiliary Material Proportions

[0121]

[0122] As shown in Table 6, the best performance of the product was obtained when the weight ratio of self-made poloxamer P-HY to cannabidiol was (5-50):1, resulting in a cannabidiol solubility of up to 7.08 mg / mL and a colloidal bundle particle size of less than 90 nm.

[0123] 4. Screening of Organic Acids: Poloxamer P-HY was used as the research object for excipient screening. An organic acid or its salt in a ratio of 1 / 10 (w / w) to cannabidiol and poloxamer P-HY (Poloxamer P-HY:CBD = 25:1 (w / w)) was added to prepare the composition according to method 1a). Under accelerated stability evaluation conditions (temperature 40℃±2℃, relative humidity 75±5%, samples taken at 0, 1, 2, 3, and 6 months), the influence of cannabidiol content (determined by HPLC external standard method) and appearance was comprehensively evaluated on the organic acid content. The results are shown in Table 7.

[0124] The method for determining cannabidiol content by high performance liquid chromatography (HPLC) is described below:

[0125] a. Chromatographic conditions and system suitability test:

[0126] An octadecylsilane-bonded silica gel column (C18, 4.6 × 150 mm, 4 μm) was used as the stationary phase, with acetonitrile-water (70:30) as the mobile phase. The detection wavelength was 210 nm, the flow rate was 1 mL / min, and the column temperature was 25 °C. The theoretical plate number, calculated based on the cannabidiol peak, should be no less than 2000.

[0127] b. Preparation of the reference solution:

[0128] Take an appropriate amount of cannabidiol reference standard, accurately weigh it, and prepare a solution containing approximately 0.01 mg per 1 ml using acetonitrile.

[0129] c. Preparation of the test solution:

[0130] Take 10 mg of sample, place it in a 1000 ml volumetric flask, add acetonitrile to the mark, shake well, and collect the filtrate to obtain the final product.

[0131] d. Measurement method:

[0132] Accurately inject 10 μl each of the reference solution and the test solution into the liquid chromatograph and record the chromatograms. Calculate the CBD content based on peak area using the external standard method.

[0133]

[0134] In the formula A sam The peak area of ​​CBD in the test sample;

[0135] V sam The dilution volume of the test solution is in ml;

[0136] A std The peak area of ​​the reference standard CBD;

[0137] W std The sample weight of the reference standard is in mg.

[0138] V std This represents the dilution volume of the reference solution, in ml.

[0139] Table 7 Results of organic acid screening

[0140]

[0141]

[0142] As shown in Table 7, the addition of organic acids or their salts can improve the stability of the cannabidiol composition to varying degrees and can prevent or delay the degradation and discoloration process. Citric acid and tartaric acid are the most effective, accelerating the maintenance of the original appearance of the finished product within 6 months and significantly reducing the degradation rate of cannabidiol itself. Vitamin C is the second most effective. Organic acid salts have similar effects, but are not as effective as organic acids. The best salt is the acid salt of tartaric acid (potassium hydrogen tartrate), followed by the potassium and sodium salts of citric acid.

[0143] 5. Screening of Organic Acid Ratios: Citric acid, tartaric acid, and potassium hydrogen tartrate were added to a mixture of cannabidiol and poloxamer P-HY (poloxamer P-HY:CBD = 25:1 (W / W)) at ratios of 1 / 1, 1 / 5, 1 / 10, 1 / 50, 1 / 100, or 1 / 500 to the cannabidiol component. A β-cyclodextrin:cannabidiol ratio of 50:1 was used as a control. The composition was prepared according to method a) in section 1. Under accelerated stability assessment conditions (temperature 40℃±2℃, relative humidity 75±5%, samples were taken at 0, 1, 2, 3, and 6 months), the ratio of organic acids in the composition was comprehensively evaluated using cannabidiol content (determined by HPLC external standard method, specific method as described above) and appearance as indicators.

[0144] Table 8 Results of organic acid ratio screening

[0145]

[0146]

[0147] As shown in Table 8, citric acid and tartaric acid exhibit the same trend, with significant effects occurring in the range of organic acid:cannabidiol (W / W) of 1-0.01. The optimal ratio is (0.01-0.1):1 for organic acid:cannabidiol, where the effect is similar to that of 0.1 when the organic acid ratio is above 0.1. The optimal ratio for organic acid salts is (0.02-0.1):1 for potassium hydrogen tartrate:cannabidiol.

[0148] Example 3: Evaluation of the biological activity of the cannabidiol composition

[0149] Two comparative studies were prepared according to the methods disclosed by GW PHARMA LTD in patent application GB2380129A and Russell Hobart Stebbins in patent application US20160143972 A1 to evaluate the technical effects of the cannabidiol composition described in this application, mainly including stability in gastric juice, bioavailability, and plasma protein binding rate.

[0150] Sample to be tested:

[0151] 1) Select the nine optimized formulations shown below from Example 2, and prepare the compositions using the method described in 1a) for subsequent experiments:

[0152] a. Prescription 1: Poloxamer P-HY:CBD:Citrate = 100:1:0.01;

[0153] b. Prescription 2: Poloxamer P-HY:CBD:Citrate = 50:1:0.01;

[0154] c. Prescription 3: Poloxamer P-HY:CBD:Citrate = 100:1:0.02;

[0155] Prescription 4: Poloxamer P-HY:CBD:Citrate = 100:1:0.1;

[0156] e. Prescription 5: Poloxamer P-HY:CBD:Citrate = 10:1:0.01;

[0157] f. Prescription 6: Poloxamer P-HY:CBD:Citrate = 5:1:0.01;

[0158] g. Prescription 7: Poloxamer P-HY:CBD:Citrate = 1:1:0.01;

[0159] h. Prescription 8: Poloxamer P-407: CBD: Citric acid = 50:1:0.01;

[0160] i. Prescription 9: Poloxamer P-188: CBD: Citric acid = 50:1:0.01.

[0161] 2) GW formula: Polyoxygenated castor oil RH40: Ethanol: CBD = 1:2:0.1.

[0162] 3) Russell's prescription: β-glucan:CBD = 15:1.

[0163] 1. To investigate the stability of raw material cannabidiol and different cannabidiol formulations in artificial gastric juice.

[0164] Experimental methods:

[0165] 1) Stability test method for artificial gastric fluid: Take 1 mol / ml dilute hydrochloric acid, dilute with water, and adjust the pH to 1.5. Add 1g of pepsin to every 100ml of liquid and mix well. Take equal volumes of 100ml of artificial gastric fluid, divide into 12 groups, and add 100mg of the composition to each group simultaneously according to the prescription. Use high-purity cannabidiol powder (100mg, 400 mesh, purity >99%) as a control. Shake at 37℃ for 0, 1, 4, 12, and 24h. Determine the cannabidiol concentration by high-performance liquid chromatography (HPLC). The control solution and the test solution are prepared according to the following steps:

[0166] Reference solution: Take an appropriate amount of cannabidiol reference standard, accurately weigh it, and prepare a solution containing approximately 0.01 mg per ml with acetonitrile.

[0167] Test solution: Mix the test sample thoroughly, accurately measure 10 ml, place it in a 100 ml volumetric flask, add acetonitrile to the mark, shake well, and collect the filtrate. Chromatographic conditions and determination methods are as described above.

[0168] Table 9. Stability of different cannabidiol preparations in artificial gastric fluid.

[0169]

[0170] Experimental results show that the composition has good stability in simulated gastric juice, and no significant degradation of cannabidiol or production of the psychoactive substance THC was observed.

[0171] 2. Investigate the pharmacokinetic properties of raw material cannabidiol and different cannabidiol combinations in rats.

[0172] After rats were administered cannabidiol (CBD) via gavage, whole blood samples were collected at different time points, plasma was separated, and the drug concentration in the plasma was determined by liquid chromatography-tandem mass spectrometry.

[0173] (1) Dosing regimen

[0174] Seventy-eight healthy SPF-grade rats, weighing 150-200g, were purchased from the Vital River Laboratory Animal Center (a laboratory animal company authorized by Charles River). They were randomly divided into 13 groups of six rats each. All groups were administered the drug by gavage. The rats were fasted for 12 hours before the experiment but had free access to water. They were fed uniformly 2 hours after drug administration.

[0175] Cannabidiol raw materials and various prescription cannabidiol compositions were dissolved in physiological saline to form a solution or suspension of 2 mg / ml (based on CBD content) for experimental use.

[0176] All test groups were administered liquid via gavage at a dose of 10 ml / kg. 1) The blank control group was administered physiological saline via gavage; 2) Cannabidiol raw material suspension was administered directly via gavage; 3) The compositions described in prescriptions 1-9, as well as the GW prescription and the Russell prescription, were administered via gavage.

[0177] (2) Blood collection time and sample processing:

[0178] Samples were collected at 10 time points: 10 min, 20 min, 40 min, 1.0 h, 80 min, 2.0 h, 4.0 h, 8.0 h, 12.0 h, and 24 h after drug administration.

[0179] At the time points specified above, 0.3 ml of venous blood was collected from the posterior venous plexus of rats, placed in heparinized test tubes, centrifuged at 11,000 rpm for 5 min, and the plasma was separated and frozen at -20°C.

[0180] (3) Sample testing and data analysis

[0181] 1) The concentration of cannabidiol in rat plasma was determined by LC / MS / MS.

[0182] The non-compartmental model of WinNonlin software was used to analyze and calculate the pharmacokinetic parameters after drug administration.

[0183] 2) Mass spectrometry conditions:

[0184] Instrumentation: AB SCIEX Triple Quad™ 5500 system; ESI: positive ion; Monitoring mode: SRM; Cannabidiol: [M+H] + m / z 315.2 / 123.2; Glibenclamide (internal standard): [M+H] + m / z 494.2 / 169.1.

[0185] 3) UPLC conditions:

[0186] Mobile phase A: 0.1% formic acid & 2mM ammonium formate aqueous-acetonitrile solution (v:v, 95:5)

[0187] Mobile phase B: 0.1% formic acid & 2mM ammonium formate acetonitrile aqueous solution (v:v, 95:5)

[0188] Table 10 UPLC Conditions

[0189] 0 20.0 0.30 20.0 0.80 90.0 1.20 90.0 1.21 20.0 1.30 20.0

[0190] 4) Chromatographic conditions:

[0191] Chromatographic column: ACQUITY UPLC Protein BEH C4 column, column temperature: 60℃, flow rate: 0.7 ml / min, retention time: cannabidiol: 0.96 min, glibenclamide (internal standard): 0.87 min;

[0192] 5) Sample preparation: Accurately take 5 μL of blood sample, add 100 μL of internal standard, vortex, centrifuge at 1300 rpm and 4℃ for 10 min, and take 10 μL of supernatant for analysis.

[0193] (4) Detection indicators

[0194] 1) Detect the plasma concentration of cannabidiol in rats after oral administration of 20 mg / kg;

[0195] 2) Detect the pharmacokinetic parameters (peak concentration C) of rats after gavage administration of cannabidiol and the cannabidiol combination. max Peak time T max The results (AUC, relative bioavailability with cannabidiol suspension as reference) are shown in Table 11.

[0196] (5) Research Results

[0197] Compared to raw cannabidiol, cannabidiol formulations 1-9 have a higher C content. max Higher, T max Shorter administration time allows for faster achievement of effective concentrations and exertion after oral administration. The AUC and relative bioavailability of formulations 1-9 were significantly better than those of the GW and Russell formulations, suggesting that the cannabidiol compositions have better absorption in vivo.

[0198] Table 11 Pharmacokinetic parameters of different cannabidiol preparations after gavage administration to mice

[0199]

[0200] 3. To investigate the therapeutic effects of raw cannabidiol and different cannabidiol formulations on a pentylenetetrazol-induced epilepsy mouse model.

[0201] (1) Experimental methods

[0202] One hundred and twenty male C57BL / 6 mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed under natural light (12 hours of light / 12 hours of darkness) conditions with free access to water and food.

[0203] Cannabidiol raw material was prepared as a 7.5 mg / ml suspension in physiological saline for experimental use; each formulation of cannabidiol composition was prepared as a 5 mg / ml aqueous solution or suspension (based on CBD content) in physiological saline for experimental use; carbamazepine was prepared as a 6.25 mg / ml aqueous solution or suspension in physiological saline for experimental use; pentylenetetrazol was prepared as a 6 mg / ml aqueous solution or suspension in physiological saline for experimental use.

[0204] After one week of acclimatization, mice were randomly divided into 12 groups according to their body weight: 1) a blank control group administered saline by gavage; 2) a raw material cannabidiol group administered cannabidiol suspension (150 mg / kg, 20 ml / kg) by gavage; 3) prescription groups 1-9 administered aqueous solutions of prescriptions 1-9 (100 mg / kg, 20 ml / kg) by gavage; 4) a positive control group administered carbamazepine solution (125 mg / kg, 20 ml / kg) by gavage. Two hours after gavage administration, each group was subcutaneously injected with pentylenetetrazol 60 mg / kg (10 ml / kg). The latency period of the first tonic-clonic seizure, the total duration of tonic-clonic seizures within 30 minutes, and the frequency of seizures were then observed and recorded. The Racine grading method was used to determine the most severe epileptic level reached by the mice within 30 minutes, evaluating the drug's effect on the occurrence and development of epilepsy. The specific Racine classification method is as follows: Type 1 epilepsy, lying still; Type 2 epilepsy, head nodding, occasional body jerking, tail stiffness; Type 3 epilepsy, single forelimb lifting, hindlimb spasm; Type 4 epilepsy, both forelimbs lifting, standing jerking; Type 5 epilepsy, standing and falling, violent jerking, large jumps, death.

[0205] (2) Experimental results:

[0206] Compared with the blank control group, the time of the first onset of tonic-clonic seizures was significantly delayed in the raw cannabidiol group and all cannabidiol composition groups except for formulation 7 (P < 0.01, P < 0.05). There were no significant differences in the total duration, frequency, and severity of tonic-clonic seizures in the raw cannabidiol group compared with the blank control group. All cannabidiol composition groups showed improvement in these three indicators compared with the blank control group. Specifically, formulations 1-6 and 8 significantly shortened the total duration and frequency of tonic-clonic seizures and reduced the severity of epileptic seizures in the model mice (P < 0.01, P < 0.05). These data indicate that, compared with raw cannabidiol, the cannabidiol compositions of this application can more effectively reduce the severity of epilepsy and delay seizures, especially the lower doses of cannabidiol compositions (formulations 1-6 and 8 with a poloxamer:CBD ratio greater than 5:1).

[0207] Table 12. Ameliorative effects of different cannabidiol preparations on pentylenetetrazol-induced epilepsy in mice.

[0208]

[0209]

[0210] #Compared with the blank control group, p < 0.05; ##Compared with the blank control group, p < 0.01

[0211] 4. To investigate the therapeutic effects of raw cannabidiol and different cannabidiol formulations on depressed mice in the forced swimming experiment.

[0212] (1) Experimental methods

[0213] One hundred and twenty Kunming mice, half male and half female, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed under natural light (12 hours of light / 12 hours of darkness) with free access to water and food.

[0214] The cannabidiol raw material was prepared as a suspension of 7.5 mg / ml in physiological saline for experimental use; each prescription cannabidiol composition was prepared as an aqueous solution or suspension of 5 mg / ml (based on CBD content) in physiological saline for experimental use; amitriptyline was prepared as an aqueous solution or suspension of 1 mg / ml in physiological saline for experimental use.

[0215] After a week of acclimatization, mice were fasted for 12-16 hours with free access to food and water, and were randomly divided into 12 groups according to their body weight: 1) Blank control group was given physiological saline by gavage; 2) Raw material cannabidiol group was given raw material cannabidiol suspension (150 mg / kg) by gavage; 3) Prescription 1-9 groups were given aqueous solutions of prescription 1-9 by gavage (100 mg / kg); 4) Positive control group was given amitriptyline solution (20 mg / kg) by gavage.

[0216] Twenty-four hours before the formal test, mice were placed in a glass cylinder (25 cm high, 10 cm in diameter) with a water depth of 10 cm and a water temperature of 24 ± 1 °C for 15 minutes of forced swimming training. After each group was administered the drug, the mice were again placed in the glass cylinder with a water depth of 10 cm and forced to swim for 6 minutes. The immobility time of the mice in the last 4 minutes was observed and recorded. The immobility time was defined as the time when the mice stopped struggling, floated in the water and remained still, or made only some necessary slight movements to keep their heads afloat. Specifically, in each cannabidiol group, the mice were placed in the glass cylinder and forced to swim for 6 minutes 2 hours after administration of cannabidiol, while in the amitriptyline group, the mice were placed in the glass cylinder and forced to swim for 6 minutes 1 hour after administration.

[0217] (2) Experimental Results

[0218] In the forced swimming test in mice, amitriptyline, raw cannabidiol, and each cannabidiol combination formulation all produced antidepressant effects. Compared with the raw cannabidiol group, the immobility time of mice in each cannabidiol combination formulation group, except for formulation 7, was further reduced (P < 0.05), indicating that lower doses of cannabidiol combinations (formulations 1-8 and 9 with a poloxamer:CBD ratio greater than 5:1) have better antidepressant effects than higher doses of raw cannabidiol.

[0219] Table 13 Effects of different cannabidiol preparations on immobility time in mice during the forced swimming test.

[0220] Blank control group 170.4±7.1 Positive control group <![CDATA[123.2±6.3 # ]]> Prescription 1 group <![CDATA[94.8±6.7 ##* ]]> Prescription 2 groups <![CDATA[83.9±4.1 ##* ]]> Prescription 3 groups <![CDATA[82.5±5.4 ##* ]]> Prescription 4 groups <![CDATA[86.1±3.9 ##* ]]> Prescription 5 groups <![CDATA[91.3±5.8 ##* ]]> Prescription 6 groups <![CDATA[97.8±7.5 ##* ]]> Prescription 7 groups <![CDATA[117.2±7.9 # ]]> Prescription 8 groups <![CDATA[93.7±4.3 ##* ]]> Prescription 9 groups <![CDATA[102.1±4.7 # ]]> Raw material cannabidiol group <![CDATA[111.5±7.7 # ]]>

[0221] #Compared with the blank control group, p < 0.05; ##Compared with the blank control group, p < 0.01; *Compared with the raw material cannabidiol group, p < 0.05

[0222] 5. Investigate the plasma protein binding rate of raw cannabidiol and different cannabidiol combination formulations.

[0223] The plasma protein binding rates of raw material cannabidiol and the cannabidiol compositions formulations 1-9 described in this application were evaluated using the commonly used balanced dialysis method.

[0224] (1) Experimental method:

[0225] 1) Sample stock solution preparation: Take raw material cannabidiol or cannabidiol composition, add methanol to prepare 6 gradients (200, 100, 50, 20, 10, 5) μg / mL. 200 μg / mL is used as high concentration sample, 100 μg / mL is used as medium concentration sample, and 50 μg / mL is used as low concentration sample.

[0226] 2) Preparation of blank dialysis solution: Accurately weigh 14.110g K2HPO4, 2.592g KH2PO4, and 1.991g NaCl and dissolve them in 950ml of deionized water. Adjust the pH to 7.4 with 0.1mol / L HCl, and then bring the volume to 1000ml.

[0227] 3) Dialysis solution: Pipette 200 μL of fresh rabbit plasma and add 600 μL of sample (cannabidiol or cannabidiol composition stock solution) to prepare the dialysis membrane solution. The dialysis membrane has a molecular weight cutoff of 5000D (model: G-RC-18-5K), and the dialysis cell is an ampoule containing 20 ml of blank dialysis solution.

[0228] 4) Specificity study: Examine the peaks of plasma (or dialysate) and plasma (or dialysate) + sample after dialysis equilibrium in the dialysis cell, and observe whether they interfere with the measurement results.

[0229] 5) Preparation of standard curve: Take 400 μL of each sample stock solution (5-100 μg / mL), add 200 μL of dialysis solution (or plasma), mix well, add 400 μL of methanol, centrifuge and take the supernatant, send it for HPLC test, and prepare a standard curve of the external dialysis solution (or internal dialysis solution) with sample concentration as the abscissa and peak area as the ordinate.

[0230] 6) Recovery rate test: 200 μL of fresh rabbit plasma was taken with a pipette and high, medium and low concentration samples were added respectively. The recovery rate of cannabidiol was determined according to the steps described in 5).

[0231] 7) Determination of plasma protein binding rate: After washing the dialysis bag with deionized water, tie one end and add three types of drug-containing plasma (200μL + 600μL) of high, medium and low concentrations respectively. After equilibration, follow the post-processing method in the standard curve, take samples and send them to HPLC for determination.

[0232] HPLC determination conditions:

[0233] An octadecylsilane-bonded silica gel column (C18, 4.6 × 150 mm, 3 μm) was used as the stationary phase, with acetonitrile-water (70:30) as the mobile phase. The detection wavelength was 210 nm, the flow rate was 1 mL / min, and the column temperature was 25 °C. The theoretical plate number, calculated based on the CBD peak, should be no less than 2000.

[0234] (2) Experimental results:

[0235] The results showed that, except for formulation 7, the plasma protein binding rates of all other formulations were significantly reduced compared to the raw material cannabidiol. This suggests that the compositions of this application help address the potential accumulation problem in cannabidiol metabolism.

[0236] Table 14 Results of plasma protein binding rate determination for different cannabidiol preparations

[0237] Raw material cannabidiol group 95 Prescription 1 79 Prescription 2 74 Prescription 3 75 Prescription 4 72 Prescription 5 78 Prescription 6 83 Prescription 7 91 Prescription 8 81 Prescription 9 89

[0238] Although specific embodiments of the invention have been described in detail, those skilled in the art will understand that various modifications and variations can be made to the details based on all the teachings disclosed, and all such changes are within the scope of protection of the invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims

1. A cannabidiol composition comprising cannabidiol, poloxamer, and an organic acid or a pharmaceutically acceptable salt thereof; in, The poloxamer has a molecular weight of 9000–16000 Da and a polyoxyethylene block percentage of 60%–80%. The organic acid is citric acid, tartaric acid, or ascorbic acid; The weight ratio of cannabidiol, poloxamer, organic acid or pharmaceutically acceptable salt thereof is 1:(5-50):(0.01-0.2).

2. The composition of claim 1, wherein the poloxamer has a molecular weight of 9000-12600 Da.

3. The composition of claim 1, wherein the poloxamer has a molecular weight of 9900 to 12600 Da.

4. The composition of claim 1, wherein the poloxamer has a molecular weight of 10,000 to 15,000 Da.

5. The composition of claim 1, wherein the poloxamer has a molecular weight of 9950 to 12600 Da.

6. The composition according to any one of claims 1-5, wherein the percentage of the poloxamer polyoxyethylene block is 60% to 75%.

7. The composition according to any one of claims 1-5, wherein the percentage of the poloxamer polyoxyethylene block is 62% to 72%.

8. The composition according to any one of claims 1-5, wherein the percentage of the poloxamer polyoxyethylene block is 65% to 75%.

9. The composition according to any one of claims 1-5, wherein the percentage of the poloxamer polyoxyethylene block is 65% to 80%.

10. The composition of claim 1, wherein the poloxamer is P407 or a poloxamer with a molecular weight of 9950-12600 Da and a polyoxyethylene block percentage of 70%.

11. The composition of claim 1, wherein the pharmaceutically acceptable salt is a sodium or potassium salt.

12. The composition of claim 1, wherein the weight ratio of cannabidiol, poloxamer, organic acid or pharmaceutically acceptable salt thereof is 1:50:0.

01.

13. The composition of claim 1, further comprising a pharmaceutically acceptable carrier or excipient.

14. The composition of claim 1, further comprising 5-20 times the weight of water in the composition's solids.

15. The composition of claim 1, further comprising 10-15 times the weight of the solids in the composition as water.

16. The composition of claim 1, further comprising 10 times the weight of the solids in the composition as water.

17. The composition of claim 14, wherein it is a nanoemulsion formulation.

18. The composition of claim 17, wherein the nanoemulsion has a particle size of 1-200 nm.

19. The composition of claim 17, wherein the nanoemulsion has a particle size of 1-100 nm.

20. The composition of claim 1, wherein it is a tablet, granule, capsule or injection.

21. The composition of claim 1, wherein it is a soft capsule.

22. Use of the composition of any one of claims 1-21 in the preparation of a medicament for treating depression or epilepsy.

23. The use according to claim 22, wherein the epilepsy is refractory epilepsy.

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