WO2019232783A1 - 大麻二酚组合物及其用途 - Google Patents

大麻二酚组合物及其用途 Download PDF

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WO2019232783A1
WO2019232783A1 PCT/CN2018/090423 CN2018090423W WO2019232783A1 WO 2019232783 A1 WO2019232783 A1 WO 2019232783A1 CN 2018090423 W CN2018090423 W CN 2018090423W WO 2019232783 A1 WO2019232783 A1 WO 2019232783A1
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cannabidiol
composition
poloxamer
improving
acid
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PCT/CN2018/090423
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English (en)
French (fr)
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张可
谭昕
于朝晖
柳旭
常坦然
金倩
高伟博
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云南汉素生物科技有限公司
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Publication of WO2019232783A1 publication Critical patent/WO2019232783A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/08—Antiepileptics; Anticonvulsants

Definitions

  • the present application relates to the field of chemistry, and in particular to a composition containing cannabidiol (CBD), which can significantly improve the water solubility of cannabidiol, making it better used in medicine, daily chemical, food, Health products and other fields.
  • CBD cannabidiol
  • Cannabis (scientific name: Cannabis sativa L.) is a family of cannabis, cannabis, also known as hemp, hemp, hemp, mountain seedlings, jute, and has important agricultural and medicinal values. Hemp contains tetrahydrocannabinol (THC), a toxic ingredient that can make people addicted and can be used as a drug. It was banned for a long time.
  • THC tetrahydrocannabinol
  • Varieties of cannabis for industrial use (referred to as “industrial cannabis”) have a THC content of less than three-thousandths in flowers and leaves during the growing period, and do not have the value of extracting the toxic component THCTHC or directly used as a drug. Economic and medicinal value, it can be legally used for large-scale planting and industrial development.
  • cannabinol compounds are a unique class of substances in cannabis plants and are the main active ingredient in cannabis plants. Research on it has been a hot topic in cannabis research.
  • the main cannabinoid compounds in cannabis plants are THC, cannabinol (CBN), CBD, cannabinol (CBG), cannabinoid (CBC), etc., the first three of which account for more than 90% of cannabinoids.
  • Cannabinol is one of the cannabinoids. Its molecular formula is C 21 H 30 O 2. It is a pale yellow resin or crystal. It is usually extracted from natural plant cannabis and can be artificially synthesized. It has many aspects of pharmacological activity. The compound is almost insoluble in water and soluble in organic solvents such as ethanol, methanol, ether, benzene, chloroform and petroleum ether. Its structural formula is shown in the following formula I:
  • Cannabidiol is completely different from tetrahydrocannabinol. It not only does not have any hallucinogenic effects, but also treats cramps, anxiety / depression, inflammation, cancer, rheumatoid arthritis, multiple sclerosis, epilepsy, especially refractory epilepsy, and Antiviral, analgesic and other aspects have good pharmacological activities, so it has become a hot spot in the global medical field.
  • cannabidiol has problems such as poor water solubility, low bioavailability, and weak stability, which greatly limits its application, as follows:
  • GW Pharma is a company whose main business is research and development of cannabis-related ingredients. Its compound drug Sativex containing cannabidiol and THC has been developed and marketed for the treatment of multiple sclerosis. However, due to the problems of poor stability and low bioavailability of gastric fluid in the administration of cannabidiol through the gastrointestinal system, it can only be developed as an oral spray and absorbed and administered through the oral mucosa. At the same time, due to the solubility of cannabidiol in the water system, the company had to choose organic solvents such as absolute ethanol and propylene glycol as a means of formulation.
  • organic solvents such as absolute ethanol and propylene glycol
  • the pharmacokinetic characteristics of different subjects showed high differences, and the coefficient of variation was large.
  • the CV% of C max for cannabinol after a single administration was 64.1%, and the CV% of AUC was 72.5 %, CV% of C max of cannabidiol after multiple consecutive administrations is 75.7%, CV% of AUC is 46.6%;
  • the present application provides a cannabidiol composition comprising cannabidiol and a surfactant.
  • the surfactant is selected from non-ionic surfactants.
  • the surfactant is selected from the group consisting of polypropylene glycol and / or polyoxyethylene as the parent adduct and polyol-type ester or amide adduct.
  • Polypropylene glycol as described herein means having The structured polymer has a molecular weight of 1000-10000 Da, preferably 5000-10000 Da, and more preferably 6000-9000 Da.
  • polyoxyethylene means The structured polymer has a molecular weight of 500-2000 Da, preferably 1000-2000 Da.
  • the polyhydric alcohol type ester adducts refer to esters formed by organic substances containing multiple hydroxyl groups in molecules such as ethylene glycol, sorbitol, and sucrose and higher fatty acids, such as sorbitol Esters, sucrose esters, etc.
  • the polyhydric alcohol amide adduct refers to an organic substance obtained by condensing a polyhydric alcohol amine (such as diethanolamine) with a fatty acid, wherein the fatty acid may be coconut oleic acid, fatty acid, or lauric acid. .
  • the surfactant is poloxamer.
  • the molecular weight of the poloxamer is 9000 to 20,000 Da, such as 9000 to 16000 Da, 9000 to 12600 Da, 9900 to 12600 Da, or 10,000 to 15000 Da, preferably 9950 to 12600 Da), and the polyoxyethylene block
  • the percentage is 40% to 80%, such as 60% to 80%, 60% to 75%, 62% to 72%, 65% to 75%, or 65% to 80%.
  • the poloxamer is P407 or a poloxamer having a molecular weight of 9950-12600 Da and a polyoxyethylene block percentage of 70%.
  • the mass ratio of cannabidiol and surfactant is 1: (1-100).
  • the mass ratio of cannabidiol and surfactant is 1: (5-50).
  • the composition also contains an antioxidant.
  • the antioxidant is an organic acid or a pharmaceutically acceptable salt thereof.
  • the antioxidant is selected from the group consisting of citric acid, tartaric acid, malic acid, succinic acid, ascorbic acid, and pharmaceutically acceptable salts thereof.
  • the antioxidant is selected from the group consisting of citric acid, tartaric acid, malic acid, succinic acid, ascorbic acid, potassium citrate, sodium citrate, and potassium hydrogen tartrate.
  • the antioxidant is citric acid or tartaric acid.
  • the weight ratio of cannabidiol and antioxidant is 1: (0.01-0.2); for example, 1: (0.01-0.1).
  • the weight ratio of cannabidiol, surfactant, and antioxidant is 1: (1-100): (0.01-0.2).
  • the weight ratio of cannabidiol, surfactant, and antioxidant is 1: (5-50): (0.01-0.1).
  • the weight ratio of cannabidiol, surfactant and antioxidant is 1: 50: 0.01.
  • the composition further contains a pharmaceutically acceptable carrier or excipient.
  • the carriers described herein include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphate, glycerol, sorbic acid, potassium sorbate, saturated plants Partial glycerin mixtures of 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, fibers Substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin.
  • the excipient refers to an additive other than the main drug in a pharmaceutical preparation. It is stable in nature, has no contraindications to the main drug, does not cause side effects, does not affect the curative effect, is not easy to deform, chapped, mildew, moth-eaten at room temperature, is harmless to the human body, has no physiological effect, and does not produce chemical or physical effects with the main drug. Effect, does not affect the content of the main drug.
  • the composition also contains 5-20 times the weight of solids; preferably 10-15 times; more preferably 10 times.
  • the solids herein refer to the remaining substances after the composition or product is sufficiently dried, that is, the sum of substances other than water in the composition.
  • the composition is a nanoemulsion formulation.
  • the nanoemulsion has a particle size of 1-200 nm.
  • the nanoemulsion has a particle size of 1-100 nm.
  • the composition is a tablet, granule, capsule, soft capsule or injection.
  • the composition may be administered orally, transdermally, or by injection.
  • the present application relates to the use of the composition in the manufacture of a medicament for the prevention and / or treatment of spasms, anxiety / depression, inflammation, cancer, rheumatoid arthritis, multiple sclerosis, epilepsy, especially refractory epilepsy , The use of antiviral drugs or analgesics.
  • the present application relates to the composition in the preparation of a food (especially a health food) that enhances immunity, assists in lowering blood glucose / lipids, aids in improving memory, clears throat, improves sleep, loses weight, or improves chemical liver damage. the use of.
  • the present application relates to the use of the composition for removing acne or improving skin moisture / oil balance in a non-therapeutic destination.
  • the application relates to the prevention and / or treatment of spasm, anxiety / depression, inflammation (e.g. rheumatoid arthritis), cancer, multiple sclerosis, epilepsy, especially refractory epilepsy, viral infection or pain
  • a method comprising the step of administering an effective amount of said composition to a subject in need thereof.
  • the present application relates to a method for enhancing immunity, assisting in lowering blood glucose / lipids, assisting in improving memory, clearing throat, improving sleep, losing weight, or improving chemical liver damage, which comprises providing a subject in need thereof A step of administering an effective amount of said composition.
  • the application relates to a method for removing acne or improving skin moisture / oil balance, comprising the step of administering an effective amount of the composition to a subject in need thereof.
  • the application relates to the composition for the prevention and / or treatment of spasms, anxiety / depression, inflammation (e.g. rheumatoid arthritis), cancer, multiple sclerosis, epilepsy, especially refractory epilepsy , Viral infection or pain.
  • the application relates to the composition for enhancing immunity, assisting in lowering blood glucose / lipids, assisting in improving memory, clearing throat, improving sleep, losing weight, or improving chemical liver damage.
  • the application relates to the composition for removing acne or improving skin moisture / oil balance.
  • the application relates to a cosmetic product comprising a composition as described above.
  • Cosmetics referred to in this article refers to spreading on any part of the human body surface, such as skin, hair, fingernails, lips, teeth, etc., by applying, spraying or other similar methods, in order to achieve cleansing, maintenance, beauty, grooming and changing the appearance, or correction Human odor, chemical industrial products or fine chemical products for the purpose of maintaining good condition.
  • the cosmetics are useful for removing acne or improving skin moisture / oil balance.
  • the present application relates to a food product (eg, a health food) that contains a composition as described above.
  • the food may be used to enhance immunity, assist in lowering blood glucose / lipids, assist in improving memory, clearing throat, improving sleep, losing weight, or improving chemical liver damage.
  • the "molecular weight” herein refers to "the number average molecular weight Mn”.
  • the term "health food” refers to foods with additional effects (usually referred to as promoting health or preventing disease) obtained by adding new nutrients or by increasing the content of certain inherent nutrients.
  • the food also has the function of regulating human functions, but it is not for the purpose of treating diseases. From the appearance point of view, it may be no different from ordinary food. For certain people, it can be used as part of daily diet.
  • the term "subject" refers to an animal, particularly a mammal, preferably a human.
  • the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain a desired effect.
  • a prophylactically effective amount refers to an amount sufficient to prevent, prevent, or delay the onset of a disease
  • a therapeutically effective amount refers to an amount sufficient to cure or at least partially prevent a disease and its complications in a patient already suffering from the disease. It is well within the ability of those skilled in the art to determine such an effective amount.
  • the amount effective for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the general condition of the patient such as age, weight and sex, the manner in which the drug is administered, and other treatments administered concurrently and many more.
  • the present application provides a cannabidiol composition, which can achieve at least one of the following technical effects:
  • composition can significantly improve the water solubility of cannabidiol, which greatly promotes its application in many fields such as medicine, daily chemical, food, health products and the like.
  • composition has high stability and solves the problems of cannabidiol instability and easy degradation in gastric juice
  • composition can significantly increase the relative bioavailability of cannabidiol
  • FIG. 1 is a GPC measurement chart of poloxamer P-HY prepared by the present application.
  • Cannabinol Yunnan Hansu Biotechnology Co., Ltd.
  • the reference standard is the end of the reflux state.
  • Ethylene oxide is condensed to both ends of the polyoxypropylene chimeric segment obtained in step (1) under the catalysis of potassium hydroxide (or sodium hydroxide) and EDTA, and the molar ratio of the starting material is controlled to be 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 ° C
  • the time 9-20h
  • a poloxamer (named P-HY) with a molecular weight ranging from 9954 to 12506 Da and a polyoxyethylene segment of 62 to 72% was obtained, and the final total yield was 45% to 50%.
  • step (1) Using the polyoxypropylene chimney segment obtained in step (1) as the raw material, add the dry inorganic alkali fine powder and EDTA according to the feeding amount in Table 2, fill with nitrogen to exhaust the air, turn on the stirring, and slowly heat in the boiling water bath. Add ethylene oxide from the bottom of the reactor and continue the reaction. The reaction time is as described in Table 2. Stop the reaction to obtain a crude yellow poloxamer copolymer. Dissolve it in purified water.
  • M sam is the molecular weight of the sample; Vsam is the volume of the sample eluent.
  • the GPC measurement spectrum data of homemade poloxamer P-HY is shown in Figure 1.
  • the poloxamer P-HY segment is at the 6th peak with a retention time of 24.948 min. It is the main material peak with a percentage area of 88.73%.
  • the average molecular weight is 11,432 Da, the molecular weight of the oligomer is mainly below 8000 Da, and the larger molecular polymer is above 15,000 Da.
  • A1 is the integrated area of the doublet at 1.15 * 10 -6 , which represents the methyl group of polyoxypropylene;
  • A2 is the integrated area of the complex peak at (3.2-3.8) * 10 -6 , and represents polyoxypropylene-based, polyoxyethylene-based CH 2 O, and polyoxypropylene-based CHO.
  • the HLB of a surfactant is represented by the difference between the sum of the HLB of each atomic group of a hydrophilic group and a lipophilic group.
  • HLB ⁇ (hydrophilic group) - ⁇ (lipophilic group) +7
  • the HLB value of oxyethylene group is 0.33, and the HLB value of oxypropylene group is 0.15.
  • the approximate HLB value of poloxamer is calculated by the formula. The results are shown in Tables 3 and 4.
  • the cannabidiol composition described in this application can be prepared by the following processes:
  • Heating and melting Put the components in the prescribed amount in a container of suitable size, and heat them above 65 ° C to make the components in a molten state. Then, stir the components sufficiently at this temperature, and then Cool to room temperature and crush the homogeneous mixture using the method described in a);
  • Spray drying Weigh out each component in the prescribed amount and disperse it in purified water at a solid content of 1 to 5%. After uniform dispersion, spray dry to powdery material through a spray dryer, and then use a) The method pulverizes the materials after spray drying;
  • dry granulation the components in the prescribed amount are placed in a dry granulator for dry granulation, and finally the granules obtained are pulverized by the method described in a);
  • Wet granulation Put the prescribed amount of each component in a wet granulator, add ethanol as a wetting agent, perform wet granulation, and then dry the prepared granules below 50 ° C to remove ethanol. Finally, the method described in a) is used for pulverization of the obtained particles.
  • the cannabidiol composition After the cannabidiol composition is dissolved in about 10 times the amount (W / W) or more of water, it is possible to form a thermodynamically stable nanoemulsion by self-emulsification.
  • the maximum solubility, the state of the solution, and the colloidal particle size can be measured by the following methods. The results are shown in Table 5.
  • Dissolved solution state Take each group of prescription samples, configure them as 2% (w / w) solution, and observe the physical form of the dissolved solution.
  • micellar particle size Wet measurement using a Mastersizer 2000 laser particle size analyzer (MS2000, 0.02-2000 ⁇ m, 0.2% shading), the sample is configured as a 0.1 mg / ml solution, filtered through a 0.45 ⁇ m filter membrane, and the initial filtrate is discarded The subsequent filtrate is the sample to be tested.
  • MS2000 Mastersizer 2000 laser particle size analyzer
  • HPLC high performance liquid chromatography
  • Octadecylsilane-bonded silica gel as filler (C18, 4.6 ⁇ 150mm, 4um column), acetonitrile-water (70:30) as mobile phase, detection wavelength is 210nm, flow rate is 1ml / min, column temperature 25 ° C.
  • the theoretical number of plates calculated from the cannabidiol peak should not be less than 2000.
  • cannabidiol reference substance Take an appropriate amount of cannabidiol reference substance, accurately weigh it, and use acetonitrile to make a solution containing about 0.01mg per 1ml.
  • a sam is the peak area of the CBD in the test product
  • V sam is the dilution volume of the test solution, ml
  • a std is the peak area of the reference CBD
  • W std is the weighing amount of the reference substance, mg
  • V std is the dilution volume of the reference solution, ml.
  • organic acids or salts can improve the stability of the cannabidiol composition to different degrees, and can avoid or delay the process of degradation and discoloration.
  • the best are citric acid and tartaric acid, which can maintain the appearance of the original finished product within 6 months of acceleration, and significantly reduce the degradation rate of cannabidiol itself.
  • the second best is vitamin C; organic acid salts have similar effects, but less than organic acids.
  • the optimal salts are acid salts of potassium tartaric acid (potassium hydrogen tartrate), and the second most preferred are potassium and sodium salts of citric acid.
  • the fraction ratio (W / W) is 1/1, 1/5, 1/10, 1/50, 1/100 or 1/500 of citric acid, tartaric acid and potassium hydrogen tartrate.
  • Phenol content determined by HPLC external standard method, the specific method is as described above
  • appearance properties are used as indicators, and the proportion of organic acids in the composition is comprehensively evaluated.
  • Stability test method of artificial gastric juice Dilute hydrochloric acid with a concentration of 1mol / ml, dilute with water, and adjust the pH to 1.5. Add 1g pepsin per 100ml of liquid and mix. Take an equal amount of 100ml artificial gastric juice and divide it into 12 groups, and then add 100mg of the composition at the same time, using high purity cannabidiol fine powder (100mg, 400 mesh, purity> 99%) as a control, and shake at 37 ° C on a constant temperature shaker. The samples were taken at 0,1,4,12,24h time points, and the concentration of cannabidiol was determined by high performance liquid chromatography (HPLC). The reference solution and the test solution are prepared by referring to the following steps:
  • Reference solution Take an appropriate amount of cannabidiol reference, accurately weigh it, and use acetonitrile to make a solution containing about 0.01mg per 1ml.
  • Test solution thoroughly mix the experimental sample, accurately measure 10ml, place it in a 100ml volumetric flask, add acetonitrile to the scale line, shake well, and take the filtrate to obtain. Chromatographic conditions and measurement methods are as described above.
  • composition has good stability in simulated gastric juice, and no significant degradation of cannabidiol and no psychoactive substance THC are found.
  • Seventy-eight healthy SPF rats were purchased from Weitonglihua Experimental Animal Center (Experimental Animal Company authorized by Charles River), weighing 150-200g, and randomly divided into 13 groups of 6 rats each. Each group was administered by gavage, fasted for 12 hours before the test, and drank freely. Eat uniformly 2h after administration.
  • the raw materials of cannabidiol and the various cannabidiol compositions were dissolved in physiological saline into a solution or suspension of 2mg / ml (based on CBD content) for experiment.
  • test group was orally administered with liquid at an administration volume of 10 ml / kg.
  • the blank control group was orally administered with physiological saline; 2)
  • the cannabinol raw material suspension was directly administered orally: 3)
  • the composition described in prescriptions 1-9 as well as the GW prescription and Russell prescription were administered orally.
  • Samples were taken at 10 time points 10min, 20min, 40min, 1.0h, 80min, 2.0h, 4.0h, 8.0h, 12.0h, and 24h after administration.
  • venous blood was taken from the venous plexus of the rat's eyeball, placed in a heparinized tube, centrifuged at 11,000 rpm for 5 min, and the plasma was separated and frozen in a -20 ° C refrigerator.
  • the non-compartment model of WinNonlin software was used to analyze and calculate the pharmacokinetic parameters after administration.
  • Mobile phase B 0.1% formic acid & 2mM ammonium formate acetonitrile-water solution (v: v, 95: 5)
  • Sample preparation Precisely take 5 ⁇ L of blood sample and add 100 ⁇ L of internal standard, vortex, 1300 rpm, centrifuge at 4 ° C for 10 min, and take 10 ⁇ L of supernatant for injection analysis.
  • cannabidiol Compared to the feedstock cannabidiol, cannabidiol higher prescription 1-9 C max, T max shorter, effective concentrations can be achieved more quickly after oral administration and play a role.
  • the AUC and relative bioavailability of prescriptions 1-9 are significantly better than those of GW prescription and Russell prescription, suggesting that the cannabidiol composition has a better absorption effect in the body.
  • mice 120 male C57BL / 6 mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. Breeding under natural light 12h light / 12h dark conditions, free drinking and feeding.
  • the raw material of cannabidiol was dissolved in a saline solution of 7.5mg / ml for experimental use; the cannabidiol composition was dissolved in an aqueous solution or suspension of 5mg / ml (based on CBD content) in normal saline.
  • carbamazepine was dissolved in a 6.25 mg / ml aqueous solution or suspension of saline for experimental purposes; pentamidine was dissolved in a normal 6 mg / ml aqueous solution or suspension of saline for experiments.
  • mice were randomly divided into 12 groups based on body weight: 1) the blank control group was given intragastrically with saline; 2) the raw cannabinol group was given intragastrically with a raw cannabidiol suspension (150mg / kg, (Medicine volume 20ml / kg); 3) the prescription 1-9 groups were administered orally with the aqueous solution of prescription 1-9 (100mg / kg, the administration volume was 20ml / kg); 4) the positive control group was administered with the carbamazepine solution orally ( 125 mg / kg, with a volume of 20 ml / kg).
  • mice After 2 hours of intragastric administration in each group, 60 mg / kg of pentylenetetrazol was administered subcutaneously (dosage volume of 10 ml / kg), and then the statistics of the incubation period of the first forced spasm in mice and the occurrence of tonic spasm in 30 minutes were started. Total duration and frequency of attacks.
  • the behavioral Racine grading method was used to count the most severe epilepsy grades achieved by mice in 30 minutes, and to evaluate the effects of drugs on the occurrence and development of epilepsy.
  • the specific Racine classification method is as follows: type 1 epilepsy, standing still; type 2 epilepsy, nodding, occasional body twitching, and stiff tail; type 3 epilepsy, single forelimb lifting, hind limb spasm; type 4 epilepsy, lifting both forelimbs, Standing tics; Type 5 epilepsy, standing down, twitching violently, jumping sharply, and dying.
  • the time at which the first tonic clonic occurred in the raw cannabinol group and each of the cannabidiol composition prescription groups except prescription 7 was significantly delayed (P ⁇ 0.01, P ⁇ 0.05).
  • the total tonic clonic duration, frequency of attacks, and level of attacks of the raw cannabinol group were not significantly different from those of the blank control group.
  • Each of the cannabidiol composition groups was compared with the blank control group on these three indicators. There are improvements, in which prescriptions 1-6, 8 can significantly reduce the total duration and frequency of tonic clonics in model mice, and reduce the level of seizures (P ⁇ 0.01, P ⁇ 0.05).
  • mice male and female, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. Breeding under natural light 12h light / 12h dark conditions, free drinking and feeding.
  • the raw material of cannabidiol was dissolved in a saline solution of 7.5mg / ml for experimental use; the cannabidiol composition was dissolved in an aqueous solution or suspension of 5mg / ml (based on CBD content) in normal saline.
  • amitriptyline was dissolved in a 1 mg / ml aqueous solution or suspension with physiological saline for experiments.
  • mice After one week of adaptive breeding, the mice were fasted for 12-16 hours and had free water. They were randomly divided into 12 groups according to body weight: 1) the blank control group was given intragastrically with saline; 2) the raw cannabinol group was given intragastrically with raw Diphenol suspension (150mg / kg); 3) The prescription 1-9 group was orally administered with an aqueous solution of prescription 1-9 (100mg / kg); 4) the positive control group was orally administered with an amitriptyline solution (20mg / kg) kg).
  • mice were placed in glass round cylinders (height 25 cm, diameter 10 cm) with a water depth of 10 cm, and the water temperature was 24 ⁇ 1 ° C for 15 minutes of forced swimming training. After each group was administered, the mice were again forced to swim in a glass cylinder with a water depth of 10 cm for 6 minutes, and the immobility time of the mice in the last 4 minutes was observed and recorded. When the mice stopped struggling, stayed floating in the water, or only made some necessary light movements to keep their heads floating on the water, the time of swimming was considered immobile; of which, each of the cannabidiol groups was administered cannabis. After 2 hours of diphenol, the mice were forced to swim in a glass round cylinder for 6 minutes, and in the amitriptyline group, mice were forced to swim in a glass round cylinder for 1 minute after 1 h.
  • the commonly used equilibrium dialysis method is used to evaluate the plasma protein binding rate of the raw cannabidiol and the cannabidiol composition prescription 1-9 of the present application.
  • Sample mother liquor configuration take raw cannabidiol or cannabidiol composition, add methanol to configure (200, 100, 50, 20, 10, 5) ⁇ g / mL total 6 gradients, and take 200 ⁇ g / mL as high concentration sample, 100 ⁇ g / mL was used as the medium concentration sample, and 50 ⁇ g / mL was used as the low concentration sample.
  • Blank dialysate configuration accurately weigh 14.110g of K 2 HPO 4 , 2.592g of KH 2 PO 4 , 1.991g of NaCl in 950ml of deionized water, adjust the pH to 7.4 with 0.1mol / L HCl, then set the volume to 1000ml.
  • Inner dialysis solution Take 200 ⁇ L of fresh rabbit plasma with a pipette gun and add 600 ⁇ L of sample (cannabinol or mother liquor of cannabinol composition) to the test solution in the dialysis membrane.
  • the cut-off molecular weight of the dialysis membrane is 5000D (model: G-RC-18-5K), and the dialysis cell is an ampoule containing 20 ml of blank dialysate.
  • Octadecylsilane-bonded silica gel as filler (C18, 4.6 ⁇ 150mm, 3um column), acetonitrile-water (70:30) as mobile phase, detection wavelength is 210nm, flow rate is 1ml / min, column temperature 25 ° C.
  • the number of theoretical plates should not be less than 2000 calculated by the CBD peak.

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Abstract

本申请涉及大麻二酚组合物及其用途。具体地，本申请涉及大麻二酚组合物，其含有大麻二酚和表面活性剂，所述组合物可显著提高大麻二酚的水溶性，使其更好地应用于医药、日化、食品、保健品等领域。

Description

大麻二酚组合物及其用途 技术领域
本申请涉及化学领域，特别涉及一种含大麻二酚(Cannabidiol,CBD)的组合物，该组合物可显著提高大麻二酚的水溶性，使其更好地应用于医药、日化、食品、保健品等领域。
背景技术
大麻(学名：Cannabis sativa L.)为大麻科、大麻属植物，又名麻、汉麻、火麻、山丝苗、黄麻，具有重要的农用及药用价值。大麻中含有一种毒性成分四氢大麻酚(THC)可使人致幻成瘾，可用作毒品，曾在相当长时期内禁种。
专供工业用途的大麻品种(简称为“工业大麻”)其生长期花叶中的THC含量小于千分之三，不具备提取毒性成分THCTHC的价值或直接作为毒品吸食，而鉴于其极高的经济和药用价值，可以合法进行规模化种植与工业化开发利用。
目前，人们已从大麻植株中分离出了500余种物质，其中大麻酚类化合物至少有86种。大麻酚类化合物是大麻植株中特有的一类物质，是大麻植物中主要的活性成分，有关它的研究一直是大麻研究的热点。大麻植株中主要的大麻酚类化合物有THC、大麻酚(CBN)、CBD、大麻萜酚(CBG)、大麻环萜酚(CBC)等，其中前三者占大麻酚类化合物的90％以上。
大麻二酚是大麻素类物质中的一种，分子式为C 21H 30O 2，是一种苍黄色树脂或结晶，通常从天然植物大麻中提取得到也可人工合成，具有多方面药理活性的化合物，几乎不溶于水，溶于乙醇、甲醇、乙醚、苯、氯仿及石油醚等有机溶剂。其结构式如下面的式I所示：
Figure PCTCN2018090423-appb-000001
大麻二酚与四氢大麻酚完全不同，它不仅无任何致幻作用，并且在治疗痉挛、焦虑/抑郁、炎症、癌症、风湿性关节炎、多发性硬化症、癫痫特别是难治性癫痫以及抗病毒、镇痛等方面具有良好的药理活性，因此，成为全球医药领域研究的热点。
然而大麻二酚存在水溶性差、生物利用度低、稳定性弱等问题，这在很大程度上限制了它的应用，具体如下：
GW Pharma是一家以研究开发大麻相关成分药物为主要业务的公司，其拥有的含大麻二酚和THC的复方药物Sativex已开发上市，应用于多发性硬化症的治疗。但是由于大麻二酚经胃肠道系统给药存在胃液中稳定性差、生物利用度低等问题，只能将其开发为口腔喷雾剂，通过口腔黏膜吸收给药。同时，也因大麻二酚在水系中的溶解性问题，该公司不得不选用无水乙醇、丙二醇这样的有机溶剂作为制剂成型的手段。
但是，根据Sativex在Summary of Product Characteristics中披露的信息，尽管选择了口腔黏膜给药、同时选择了有机溶剂体系，Sativex仍存在以下问题：
1)药代动力学行为受进食与否影响非常大，空腹和进食条件下相比，大麻二酚的C max和AUC分别相差了3.3倍和5.1倍；
2)在不同受试者体内，表现出较高的药代动力学特点差异，变异系数很大，单次给药后大麻二酚的C max的CV％为64.1％、AUC的CV％为72.5％，多次连续给药后大麻二酚的C max的CV％为75.7％、AUC的CV％为46.6％；
3)在同一受试者体内，也同样表现出高度的药代动力学行为变异性。
这些问题的存在与大麻二酚本身的特性有关，当然，与Sativex选择的这一特殊的给药途径和所选用的制剂技术也有很大关系。综合而言，这些巨大的不确定性，给临床上带来了有效性的显著差异，同时，也不可避免的带来一定的安全隐患。
Russell Hobart Stebbins在专利US20160143972 A1中，披露了一种制备固态大麻二酚的方法，声称这种形式的大麻二酚能够溶解于水相体系。尽管上述现有技术一定程度上改善了大麻二酚在水中溶解性差、生物利用度低的问题，但是效果差强人意。
另外，需要进一步指出的是，除了水溶性差，关于大麻二酚的安全性、顺应性、经济性、稳定性方面的问题也是无法回避的：
1)大麻二酚生物利用度低，需要通过大剂量给药来达到有效性的目标，但是，大剂量的使用必然带来顺应性问题以及增加患者经济负担，同时大剂量服用药物会也加重肝 肾负担，导致不可预测的肝肾疾病；
2)大麻二酚具有高脂溶性(K油-水＝6-7)，表观分布容积约为32L/kg，可迅速分布至大脑、脂肪组织和其他器官中，大麻二酚血浆蛋白结合率高，且约10％可与红细胞结合，长期服用在患者体内蓄积的可能性大，尤其对于肥胖的患者；
3)John Merrick,Brian Lane等在Identification of Psychoactive Degradants of Cannabidiol in Simulated Gastric and Physiological Fluid一文中公开了大麻二酚在模拟人工胃液中的情况，指出大麻二酚在该实验条件下，60分钟降解掉了85％、在120分钟时有98％被降解掉，这是非常不利的，更为可怕的是，大麻二酚在人工胃液降解后，转化成的物质是精神活性物质四氢大麻酚，这是十分危险的，并且实际应用中必然带来巨大的安全隐患。
综上可知，寻找合适的技术手段解决大麻二酚在应用中存在的问题成为本领域亟待解决的难题。
发明内容
因此，一方面，本申请提供一种大麻二酚组合物，其含有大麻二酚和表面活性剂。
在某些优选的实施方案中，所述表面活性剂选自非离子型表面活性剂。
在某些优选的实施方案中，所述表面活性剂选自聚丙二醇和/或聚氧乙烯为母体的加成物以及多元醇型的酯类或酰胺加成物。
本文中所述聚丙二醇是指具有
Figure PCTCN2018090423-appb-000002
结构的聚合物，其分子量为1000-10000Da，优选5000-10000Da，更优选6000-9000Da。
本文中所述聚氧乙烯是指具有
Figure PCTCN2018090423-appb-000003
结构的聚合物，其分子量为500-2000Da，优选1000-2000Da。
在某些优选的实施方案中，所述多元醇型的酯类加成物是指乙二醇、山梨醇、和蔗糖等分子中含有多个羟基的有机物与高级脂肪酸形成的酯，例如山梨醇酯、蔗糖酯等。
在某些优选的实施方案中，所述多元醇型酰胺加成物是指多元醇胺(例如二乙醇胺)与脂肪酸缩合而成的有机物，其中所述脂肪酸可为椰子油酸、脂肪酸或月桂酸。
在某些优选的实施方案中，所述表面活性剂为泊洛沙姆。
在某些优选的实施方案中，所述泊洛沙姆分子量为9000～20000Da，例如9000～ 16000Da、9000～12600Da、9900～12600Da或10000～15000Da，优选9950～12600Da)，并且聚氧乙烯嵌段百分比为40％～80％,例如60％～80％、60％～75％、62％～72％、65％～75％或65％～80％。
在某些优选的实施方案中，所述泊洛沙姆为P407或分子量为9950～12600Da并且聚氧乙烯嵌段百分比为70％的泊洛沙姆。
在某些优选的实施方案中，所述大麻二酚和表面活性剂的质量比为1：(1-100)。
在某些优选的实施方案中，所述大麻二酚和表面活性剂的质量比为1：(5-50)。
在某些优选的实施方案中，所述组合物中还含有抗氧化剂。
在某些优选的实施方案中，所述抗氧化剂为有机酸或其药学上可接受的盐。
在某些优选的实施方案中，所述抗氧化剂选自柠檬酸、酒石酸、苹果酸、琥珀酸、抗坏血酸及其药学上可接受的盐。
在某些优选的实施方案中，所述抗氧化剂选自柠檬酸、酒石酸、苹果酸、琥珀酸、抗坏血酸、柠檬酸钾、柠檬酸钠和酒石酸氢钾。
在某些优选的实施方案中，所述抗氧化剂为柠檬酸或酒石酸。
在某些优选的实施方案中，所述大麻二酚和抗氧化剂的重量比为1:(0.01-0.2)；例如1:(0.01-0.1)。
在某些优选的实施方案中，所述大麻二酚、表面活性剂和抗氧化剂的重量比为1:(1-100):(0.01-0.2)。
在某些优选的实施方案中，所述大麻二酚、表面活性剂和抗氧化剂的重量比为1:(5-50):(0.01-0.1)。
在某些优选的实施方案中，所述大麻二酚、表面活性剂和抗氧化剂的重量比为1：50：0.01。
在某些优选的实施方案中，所述组合物中进一步含有药学上可接受的载体或赋形剂。
这里所述的载体包括但不限于：离子交换剂，氧化铝，硬脂酸铝，卵磷脂，血清蛋白如人血白蛋白，缓冲物质如磷酸盐，甘油，山梨酸，山梨酸钾，饱和植物脂肪酸的部分甘油酯混合物，水，盐或电解质，如硫酸鱼精蛋白，磷酸氢二钠，磷酸氢钾，氯化钠，锌盐，胶态氧化硅，三硅酸镁，聚乙烯吡咯烷酮，纤维素物质，聚乙二醇，羧甲基纤维素钠，聚丙烯酸酯，蜂蜡，羊毛脂。所述赋形剂是指在药物制剂中除主药以外的附加物。其性质稳定，与主药无配伍禁忌，不产生副作用，不影响疗效，在常 温下不易变形、干裂、霉变、虫蛀、对人体无害、无生理作用，不与主药产生化学或物理作用，不影响主药的含量测定等。如片剂中的黏合剂、填充剂、崩解剂、润滑剂；中药丸剂中的酒、醋、药汁等；半固体制剂软膏剂、霜剂中的基质部分；液体制剂中的防腐剂、抗氧剂、矫味剂、芳香剂、助溶剂、乳化剂、增溶剂、渗透压调节剂、着色剂等均可称为赋形剂。
在某些优选的实施方案中，所述组合物中还含有固形物5-20倍重量的水；优选10-15倍；更优选10倍。
本文中所述固形物是指所述组合物或产品经充分干燥后，余下的物质，也即所述组合物中除水外的物质的总和。
在某些优选的实施方案中，所述组合物为纳米乳制剂。
在某些优选的实施方案中，所述纳米乳的粒径为1-200nm。
在某些优选的实施方案中，所述纳米乳的粒径为1-100nm。
在某些优选的实施方案中，所述组合物为片剂、颗粒剂、胶囊、软胶囊或注射剂。
在某些优选的实施方案中，所述组合物可通过口服、透皮或注射给药。
在另一个方面，本申请涉及所述组合物在制备用于预防和/或治疗痉挛、焦虑/抑郁、炎症、癌症、风湿性关节炎、多发性硬化症、癫痫特别是难治性癫痫的药物、抗病毒药物或镇痛药物中的用途。
在另一个方面，本申请涉及所述组合物在制备增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤的食品(尤其是保健食品)中的用途。
在另一个方面，本申请涉及所述组合物在以非治疗目的地祛痤疮或改善皮肤水分/油分平衡中的用途。
在另一个方面，本申请涉及一种预防和/或治疗痉挛、焦虑/抑郁、炎症(例如风湿性关节炎)、癌症、多发性硬化症、癫痫特别是难治性癫痫、病毒感染或疼痛的方法，其包括向有此需要的受试者施用有效量的所述组合物的步骤。
在另一个方面，本申请涉及一种增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤的方法，其包括向有此需要的受试者施用有效量的所述组合物的步骤。
在另一个方面，本申请涉及一种祛痤疮或改善皮肤水分/油分平衡的方法，其包括向有此需要的受试者施用有效量的所述组合物的步骤。
在另一个方面，本申请涉及所述组合物，其用于预防和/或治疗痉挛、焦虑/抑郁、炎症(例如风湿性关节炎)、癌症、多发性硬化症、癫痫特别是难治性癫痫、病毒感染或疼痛。
在另一个方面，本申请涉及所述组合物，其用于增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤。
在另一个方面，本申请涉及所述组合物，其用于祛痤疮或改善皮肤水分/油分平衡。
在另一个方面，本申请涉及一种化妆品，其含有前文所述的组合物。
本文中所述化妆品是指以涂抹、喷洒或者其他类似方法，散布于人体表面的任何部位，如皮肤、毛发、指趾甲、唇齿等，以达到清洁、保养、美容、修饰和改变外观，或者修正人体气味，保持良好状态为目的的化学工业品或精细化工产品。
在某些实施方案中，所述化妆品可用于祛痤疮或改善皮肤水分/油分平衡。
在另一个方面，本申请涉及一种食品(例如保健食品)，其含有前文所述的组合物。
在某些优选的实施方案中，所述食品可用于增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤。
如无特别说明，本文中所述“分子量”特指“数均分子量Mn”。
如本文中所使用的，术语“保健食品”是指通过加入新的营养成分或通过提高某些固有营养成分的含量而获得的具有附加功效(通常指促进健康或预防疾病)的食品。该食品除满足基本营养需求外，还具有调节人体功能作用，但不以疾病治疗为目的， 其从外观上来看可能与普通食品无异，对于特定人群而言，其可作为日常饮食的一部分。
如本文中所使用的，术语“受试者”是指动物，特别是哺乳动物，优选人。
如本文中所使用的，术语“有效量”是指，足以获得或至少部分获得期望的效果的量。例如，预防有效量是指，足以预防，阻止，或延迟疾病的发生的量；治疗有效量是指，足以治愈或至少部分阻止已患有疾病的患者的疾病和其并发症的量。测定这样的有效量完全在本领域技术人员的能力范围之内。例如，对于治疗用途有效的量将取决于待治疗的疾病的严重度，患者自己的免疫系统的总体状态，患者的一般情况例如年龄、体重和性别，药物的施用方式，以及同时施用的其他治疗等等。
发明的有益效果
本申请提供一种大麻二酚组合物，其能够实现下述至少一种技术效果：
1)所述组合物能够显著提高大麻二酚的水溶性，极大地推动了其在医药、日化、食品、保健品等诸多领域的应用。
2)所述组合物具有较高稳定性，解决了大麻二酚在胃液中不稳定、易降解的问题；
3)所述组合物能够显著提高大麻二酚的相对生物利用度；
4)大麻二酚在体内的药代行为更加一致，不同个体间的C max和AUC两项药代关键指标的变异系数较低，降低了临床应用过程中的不可预测的风险；
5)所述组合物中大麻二酚与血浆蛋白的结合率同单独给予大麻二酚相比，大大下降，解决了公开报道的，大麻二酚因脂溶性高，血浆蛋白结合率高，长期使用存在蓄积的问题。
附图说明
图1为本申请制得的泊洛沙姆P-HY的GPC测定图谱。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注 明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
大麻二酚：云南汉素生物科技有限公司。
实施例1：自制泊洛沙姆P-HY的制备及性能参数测定
1、泊洛沙姆(P-HY)的合成工艺路线：
1)
Figure PCTCN2018090423-appb-000004
2)
Figure PCTCN2018090423-appb-000005
(1)将氧化丙烯缩合到1,2-丙二醇基上，得聚氧丙烯嵌合段；其中碱性的1,2-丙二醇做为诱发剂，各物质摩尔比为氧化丙烯：1，2-丙二醇：KOH/NaOH＝(220-320)：1：(2-0.5)，反应温度为90-120℃，反应终点(时间)以不同的反应条件而异，参照标准是回流状态的结束。
(2)将氧化乙烯在氢氧化钾(或氢氧化钠)和EDTA的催化下，缩合到步骤(1)所得聚氧丙烯嵌合段的两端，控制起始物料的摩尔比例为氧化乙烯：聚氧丙烯：氢氧化钾(或氢氧化钠)：EDTA＝(450-650)：1：(0.5-1)：(0.05-0.1)，反应温度为90-120℃、时间为9-20h，最终得分子量范围9954～12506Da、聚氧乙烯段62～72％的泊洛沙姆(命名为P-HY)，最终总收率为45％-50％。
2、泊洛沙姆P-HY的具体制备方法
(1)聚氧丙烯嵌合段的制备
按表1物料配比投料反应。首先，在反应釜中依次加入无水硫酸钠脱水的丙二醇作为诱发剂，以及减压干燥的KOH/NaOH细粉(80-100目)，充氮排尽空气，开启搅拌， 并通入热水(95～100℃)进行加热，当KOH/NaOH完全溶解，缓缓加入氧化丙烯，反应开始进行，当反应釜热回流现象结束，即停止供应热水，以自来水循环降温，即得分子量3500Da左右的黄色粘液状聚氧丙烯粗品，将此粗品加等量水，以2倍体积正庚烷萃取，反复以纯化水洗涤提纯，最终减压浓缩，得到黄色油状聚氧丙烯嵌合段。
表1 制备聚氧丙烯嵌合段的投料与产出表
Figure PCTCN2018090423-appb-000006
(2)泊洛沙姆P-HY的制备
以步骤(1)中得到的聚氧丙烯嵌合段为原料，按表2中投料量加入干燥的无机碱细粉和EDTA，充氮排尽空气，开启搅拌，在沸水浴加热下，缓缓从反应釜底部加入环氧乙烷，继续反应，反应时间如表2中所述，停止反应即得淡黄色糊状的泊洛沙姆共聚体粗品，以纯化水溶解，滴加1mol/L的HCl，调至pH 6.5-7.0，以300-500目中性氧化铝过滤，除去大分子聚合物，滤液上大孔树脂D101柱，以2倍柱体积纯化水冲柱除盐和低聚物，冲3倍柱体积的15％乙醇得目标产物段洗脱物。将目标产物段洗脱物在60℃减压浓缩至稠膏，倒出，置于40℃，-0.10Mpa真空干燥48h，得水分0.2％以下的泊洛沙姆P-HY成品。
表2 制备泊洛沙姆PH-Y的投料与产出表
Figure PCTCN2018090423-appb-000007
3、泊洛沙姆P-HY与市售各型号泊洛沙姆性能参数对比
(1)GPC法测定泊洛沙姆的分子量
取各泊洛沙姆样品配置成5％的溶液，使用Waters 1515GPC凝胶色谱仪测定，色谱柱为G5000PWXL柱(10μm，7.8*300mm)，以四氢呋喃为流动相，25℃，流速1ml/min，256nm波长(检测器型号：Waters 2489)下检测。
以已知分子量的聚氧丙烯(分子量：3890Da)为标品所得校准曲线为：
Log M sam＝9.03-0.19Vsam(R 2＝0.998)。
M sam为样品分子量；Vsam为样品洗脱液体积。
自制泊洛沙姆P-HY的GPC测定谱图数据详见图1：泊洛沙姆P-HY段在第6个峰，保留时间为24.948min，为主要物质峰，百分面积为88.73％，平均分子量为11432Da，寡聚物分子量主要在8000Da以下，较大分子聚合物在15000Da以上。
各市售泊洛沙姆样品分子量测定结果见表3和表4。
(2)泊洛沙姆聚氧乙烯嵌合段含量测定：
取各泊洛沙姆样品，分别用含1％四甲基硅烷的氘代甲醇(或重水)溶液溶解成10％-20％(g/ml)溶液，取0.5-1.0ml上述溶液装入NMR管中，加1滴重水，振摇，在NMR仪中，从0*10 -6到5*10 -6扫描，以直接比较法定量，按下式计算PEO值，得聚氧乙烯基在整个分子组成中所占的比例，结果见表3和表4：
PEO％＝3300a/(33a+58)
式中a＝(A2/A1)-1
A1为1.15*10 -6处双峰的积分面积，代表聚氧丙烯的甲基；
A2为(3.2-3.8)*10 -6处复合峰的积分面积，代表聚氧丙烯基、聚氧乙烯基的CH 2O和聚氧丙烯基的CHO。
(3)亲水亲油平衡值(HLB值)计算：
通过用亲水基和亲油基的各个原子团的HLB总和的差值，来表示表面活性剂的HLB。
HLB＝Σ(亲水基)-Σ(亲油基)+7
其中氧乙烯基的HLB值为0.33，氧丙烯基的HLB值为0.15，通过公式计算泊洛沙姆 的HLB近似值，结果见表3和表4。
(4)pH值测定：将各泊洛沙姆样品分别配置成2％(W/W)的稀溶液，3个平行组，以精密pH计(型号：梅特勒S220)测定，取均值得pH值，结果见表3。
(5)不同配方的大麻二酚样品溶解度测定：取各待测泊洛沙姆样品与自制泊洛沙姆P-HY，与大麻二酚以25：1的重量比混匀，制成半成品，测定其在室温(20-25℃)下，100ml纯化水中的最大溶解量。摇床震荡助溶，以沉淀部分6-8h内不溶解为判定终点，结果见表3。
表3 泊洛沙姆P-HY成品性能对照表
Figure PCTCN2018090423-appb-000008
表4 泊洛沙姆规格与理化性能比较
  HLB值 分子量 氧化乙烯分子数(a值) 聚氧乙烯段PEO百分比 物理状态
泊洛沙姆P124 16 2090-2360 12 47.66％ 液体
泊洛沙姆P188 29 8423-10568 80 81.53％ 固体
泊洛沙姆P215 16.5 3840-5168 47 52.14％ 半固态
泊洛沙姆P237 24 6840-8830 64 72.98％ 固体
泊洛沙姆P338 27 13680-18788 141 84.36％ 固体
泊洛沙姆P407 21.5 10890-15224 101 74.28％ 固体
泊洛沙姆P-HY 19.5 9954-12506 90 68.98％ 固体
实施例2：大麻二酚组合物的制备及性能测定
1、大麻二酚组合物制备工艺
本申请中所述大麻二酚组合物可选用以下工艺制备：
a)物理研磨：称取处方量的所述各组分，置于适合大小研钵中或者研磨专用的机器设备中，充分研磨至处方中所述各组分混合均匀，而后将混合均匀的物料用物理或者气流粉碎或者过药典筛的方式进行粉碎；
b)加热熔融：将所述处方量的各组分，置于适合大小的容器中，加热至65℃以上，使处方各组分呈现熔融的状态，然后，在此温度下充分搅拌均匀，而后冷却至室温，采用a)中所述方法进行均匀混合物的粉碎；
c)喷雾干燥：称取处方量的所述各组分，按1～5％的固含量分散于纯化水中，分散均匀后，经过喷雾干燥机，喷雾干燥为粉末状物料，而后采用a)中所述方法进行喷雾干燥后物料的粉碎；
d)干法制粒：将所述处方量的各组分，置于干法制粒机中进行干法制粒，最后采用a)中所述方法进行所得颗粒的粉碎即得；
e)湿法制粒：将所述处方量的各组分，置于湿法制粒机中，加入乙醇作为润湿剂，进行湿法制粒，然后将制得的颗粒在50℃以下干燥除去乙醇，最后采用a)中所述方法进行所得颗粒的粉碎。
2、辅料种类的筛选：按泊洛沙姆和大麻二酚重量比为50:1的规格，参照a)法制备大麻二酚组合物，以其最大溶解度、溶解液状态、胶体粒径大小为指标综合评估。
大麻二酚组合物在约10倍量(W/W)以上的水中溶解后，能够通过自乳化形成热力学稳定的纳米乳。其最大溶解度、溶解液状态和胶体粒径可通过如下方法进行测定，结果见表5。
1)最大溶解度测定方法：由于泊洛沙姆独特的溶解特性(例如，低温更易溶解，而且形成的组合物具有类似缓释的效果等)，其最大溶解度的测定方法也不同于药典常规要求。在室温(20-25℃)下，100ml纯化水中，按0.05g递增(可先快后慢)逐渐加入组合物以摇床震荡助溶，以沉淀部分过夜(6-8h)内不溶解为判定终点。
2)溶解液状态：分别取各组处方样品，配置为2％(w/w)的溶液，观察溶解液的物理形态。
3)胶束粒径测定：使用Mastersizer 2000激光粒度仪(MS2000，0.02-2000μm，0.2％遮光度)湿法测量，样品配置成0.1mg/ml溶液，以0.45μm滤膜过滤，初始滤液弃去， 续滤液即为待测样品。
表5 辅料种类筛选结果
Figure PCTCN2018090423-appb-000009
由表5中结果可知，自制P-HY(对应分子量9950-12500Da，聚氧乙烷段比例62-72％)组大麻二酚组合物的溶解度、溶解液色泽和自乳化形成的纳米乳粒径大小(＜100)均显著优于其他组。
3、辅料配比的筛选：以泊洛沙姆P-HY和P407为优选对象，依次按照泊洛沙姆：大麻二酚重量比分别为100:1、50:1、10:1、5:1和1:1按1中a)方法制备大麻二酚组合物，以其最大溶解度、溶解液状态、胶体粒径大小为指标综合评估，结果见表6。
表6 辅料配比筛选结果
Figure PCTCN2018090423-appb-000010
Figure PCTCN2018090423-appb-000011
由表6中结果可知，自制泊洛沙姆P-HY与大麻二酚重量比为(5-50)：1时，所得成品性能最佳，使大麻二酚溶解度最高可达7.08mg/mL，且形成的胶体束粒径均在90nm以下。
4、有机酸种类筛选：辅料筛选以泊洛沙姆P-HY为研究对象。将大麻二酚和泊洛沙姆P-HY的混合物(泊洛沙姆P-HY：CBD＝25：1(W/W))中，加入与大麻二酚比例为1/10(W/W)的有机酸或其盐类按1中a)方法制备组合物，在加速稳定性评估条件(温度40℃±2℃、相对湿度75±5％，于0、1、2、3、6个月取样)下，以大麻二酚含量(HPLC外标法测定)与外观性状为指标，对有机酸的影响进行综合评价，结果见表7。
其中，高效液相色谱(HPLC)测定大麻二酚含量的方法如下所述：
a.色谱条件与系统适用性试验:
以十八烷基硅烷键合硅胶为填充剂(C18，4.6×150mm，4um色谱柱)，以乙腈-水(70:30)为流动相，检测波长为210nm，流速为1ml/min，柱温为25℃。理论板数按大麻二酚峰计算应不低于2000。
b.对照品溶液的制备：
取大麻二酚对照品适量，精密称定，用乙腈制成每1ml约含0.01mg的溶液，即得。
c.供试品溶液的准备：
取样品10mg，置1000ml容量瓶，加乙腈至刻度线，摇匀，取续滤液，即得。
d.测定法：
精密量取对照品溶液和供试品溶液各10μl注入液相色谱仪，记录色谱图。按外标法，以峰面积计算CBD含量。
Figure PCTCN2018090423-appb-000012
式中A sam为供试品中CBD的峰面积；
V sam为供试品溶液的稀释体积，ml；
A std为对照品CBD的峰面积；
W std为对照品的称样量，mg；
V std为对照品溶液的稀释体积，ml。
表7 有机酸种类筛选结果
Figure PCTCN2018090423-appb-000013
由表7中结果可知，加入有机酸或其盐，能够使大麻二酚组合物稳定性得到不同程度的改善，并能避免或延缓降解变色的进程。其中，最优为柠檬酸和酒石酸，能在加速6月内保持原有成品外观，并明显降低了大麻二酚本身降解速率，次优为维生素C；有机酸盐有类似效果，但不及有机酸，最优盐类为酒石酸的酸式盐(酒石酸氢钾)，次优为柠檬酸的钾盐和钠盐。
5、有机酸配比筛选：将大麻二酚和泊洛沙姆P-HY的混合物(泊洛沙姆P-HY：CBD ＝25：1(W/W))中，分别加入与大麻二酚组分比例(W/W)为1/1、1/5、1/10、1/50、1/100或1/500的柠檬酸、酒石酸以及酒石酸氢钾。以β-环糊精：大麻二酚＝50:1的配比为对照。按1中a)方法制备组合物，在加速稳定性评估条件(温度40℃±2℃、相对湿度75±5％，于0、1、2、3、6个月取样)下，以大麻二酚含量(HPLC外标法测定，具体方法如前所述)与外观性状为指标，对组合物中有机酸的比例进行综合评价。
表8 有机酸配比筛选结果
Figure PCTCN2018090423-appb-000014
Figure PCTCN2018090423-appb-000015
由表8中结果可知，柠檬酸与酒石酸趋势一致，在有机酸：大麻二酚(W/W)为1-0.01范围内发生显著作用，最优配比为有机酸：大麻二酚为(0.01-0.1)：1，其中有机酸在0.1以上时，作用与0.1时近似；有机酸盐最优配比为酒石酸氢钾：大麻二酚为(0.02-0.1)：1。
实施例3：大麻二酚组合物的生物学活性评价
根据GW PHARMA LTD在专利申请GB2380129A中，以及Russell Hobart Stebbins在专利申请US20160143972 A1中所公开的方法，制备两种对比研究用的样品，用于评价本申请所述大麻二酚组合物的技术效果，主要包括胃液中稳定性、生物利用度、血浆蛋白结合率等方面。
待测样品：
1)选取实例2中如下所示的九组优化的配方组成，以1中a)所述方法制备组合物进行后续实验：
a.处方1：泊洛沙姆P-HY：CBD：柠檬酸＝100：1：0.01；
b.处方2：泊洛沙姆P-HY：CBD：柠檬酸＝50：1：0.01；
c.处方3：泊洛沙姆P-HY：CBD：柠檬酸＝100：1：0.02；
d处方4：泊洛沙姆P-HY：CBD：柠檬酸＝100：1：0.1；
e.处方5：泊洛沙姆P-HY：CBD：柠檬酸＝10：1：0.01；
f.处方6：泊洛沙姆P-HY：CBD：柠檬酸＝5：1：0.01；
g.处方7：泊洛沙姆P-HY：CBD：柠檬酸＝1：1：0.01；
h.处方8：泊洛沙姆P-407：CBD：柠檬酸＝50：1：0.01；
i.处方9：泊洛沙姆P-188：CBD：柠檬酸＝50：1：0.01。
2)GW处方：聚氧氢化蓖麻油RH40：乙醇：CBD＝1：2：0.1。
3)Russell处方：β-葡聚糖：CBD＝15：1。
1、考察原料大麻二酚及不同大麻二酚组合物处方在人工胃液中的稳定性。
实验方法：
1)人工胃液稳定性实验方法：取浓度为1mol/ml的稀盐酸，加水稀释，将pH调至1.5。每100ml液体中加入1g胃蛋白酶，混匀。取等量100ml人工胃液，分为12组，依次按各处方同时加入组合物100mg，以高纯大麻二酚微粉(100mg，400目，纯度＞99％)为对照，在37℃恒温摇床震荡，于0,1,4,12,24h时间点取样，高效液相色谱(HPLC)测定大麻二酚浓度。其中，对照品溶液和供试品溶液参照以下步骤制备：
对照品溶液：取大麻二酚对照品适量，精密称定，用乙腈制成每1ml约含0.01mg的溶液，即得。
供试品溶液：将实验样品充分混匀，精密量取10ml，置100ml容量瓶中，加乙腈至刻度线，摇匀，取续滤液，即得。色谱条件和测定方法如前所述。
表9 不同大麻二酚制剂在人工胃液中的稳定性
Figure PCTCN2018090423-appb-000016
实验结果表明：所述组合物在模拟胃液中的稳定性良好，未发现大麻二酚的明显降 解和精神活性物质THC的产生。
2、考察原料大麻二酚及不同大麻二酚组合物处方在大鼠体内的药代动力学性质。
大鼠灌胃给予大麻二酚待测样品后，采集不同时间的全血样品，分离血浆，以液相色谱-串联质谱法测定血浆中的药物浓度。
(1)给药方案
健康SPF级大鼠78只购自维通利华实验动物中心(Charles River授权的实验动物公司)，体重150-200g，随机分成13组，每组6只。各组灌胃给药处理，试验前禁食12h，自由饮水。给药后2h统一进食。
大麻二酚原料及各处方大麻二酚组合物均以生理盐水溶解为2mg/ml(以CBD含量计)的溶液或混悬液供实验用。
各受试组均按照10ml/kg的给药体积灌胃给予液体。1)空白对照组灌胃给予生理盐水；2)大麻二酚原料混悬直接灌胃给药：3)处方1-9所述组合物以及GW处方和Russell处方均灌胃给药。
(2)采血时间点及样品处理：
在给药后10min，20min，40min，1.0h，80min，2.0h，4.0h，8.0h，12.0h和24h,分10个时间点采样。
在以上设定的时间点经大鼠眼球后静脉丛取静脉血0.3ml，置肝素化试管中，11000rpm离心5min，分离血浆，于-20℃冰箱中冷冻。
(3)样品测试和数据分析
1)采用LC/MS/MS法测定大鼠血浆中大麻二酚浓度。
采用WinNonlin软件的非房室模型分析计算给药后的药代动力学参数。
2)质谱条件：
仪器：AB SCIEX Triple QuadTM 5500系统；ESI：正离子；监测模式：SRM；大麻二酚:[M+H] +m/z 315.2/123.2；优降糖(内标):[M+H] +m/z 494.2/169.1.
3)UPLC条件：
流动相A:0.1％甲酸&2mM甲酸铵水-乙腈溶液(v:v，95:5)
流动相B:0.1％甲酸&2mM甲酸铵乙腈-水溶液(v:v，95:5)
表10 UPLC条件
时间(min) 流动相B(％)
0 20.0
0.30 20.0
0.80 90.0
1.20 90.0
1.21 20.0
1.30 20.0
4)色谱条件：
色谱柱：ACQUITY UPLC Protein BEH C4柱，柱温：60℃，流速：0.7ml/min，保留时间：大麻二酚:0.96min，优降糖(内标)：0.87min；
5)样品配制：精密取血样5μL加入100μL内标，涡旋振荡，1300rpm，4℃离心10min，取10μL上清液进样分析。
(4)检测指标
1)检测大鼠灌胃给药20mg/kg大麻二酚后在血浆中的浓度；
2)检测大鼠灌胃给药大麻二酚和大麻二酚组合物后的药物动力学参数(达峰浓度C max、达峰时间T max、药时曲线下面积AUC、以大麻二酚原料混悬液为参比制剂的相对生物利用度)，结果如表11所示。
(5)研究结果
与原料大麻二酚相比，大麻二酚处方1-9的C max更高，T max更短，口服给药后可更快达到有效浓度并发挥作用。处方1-9的AUC及相对生物利用度均明显优于GW处方和Russell处方，提示大麻二酚组合物在体内有更好的吸收效果。
表11 不同大麻二酚制剂对小鼠灌胃给药后的药代动力学参数
Figure PCTCN2018090423-appb-000017
Figure PCTCN2018090423-appb-000018
3、考察原料大麻二酚及不同大麻二酚组合物处方对戊四唑诱导的癫痫小鼠模型的治疗效果。
(1)实验方法
120只雄性C57BL/6小鼠，购自北京维通利华实验动物技术有限公司。自然光12h光照/12h黑暗条件下饲养，自由饮水、摄食。
大麻二酚原料以生理盐水溶解为7.5mg/ml的混悬液供实验用；各处方大麻二酚组合物均以生理盐水溶解为5mg/ml(以CBD含量计)的水溶液或混悬液供实验用；卡马西平以生理盐水溶解为6.25mg/ml的水溶液或混悬液供实验用；戊四唑以生理盐水溶解为6mg/ml的水溶液或混悬液供实验用。
小鼠适应性饲养一周后，根据体重随机分为12组：1)空白对照组灌胃给予生理盐水；2)原料大麻二酚组灌胃给予原料大麻二酚混悬液(150mg/kg，给药容积20ml/kg)；3)处方1-9组分别灌胃给予处方1-9的水溶液(100mg/kg，给药容积20ml/kg)；4)阳性对照组灌胃给予卡马西平溶液(125mg/kg，给药容积20ml/kg)。各组灌胃给药2h后，皮下注射戊四唑60mg/kg(给药容积10ml/kg)，随后开始观察统计小鼠第一次发生强制性痉挛的潜伏期以及小鼠30min内发生强直性痉挛的总持续时间及发作频率。应 用行为学Racine分级方法，统计小鼠在30min达到的最严重癫痫等级，评判药物对癫痫发生和发展的影响。具体的Racine分级方法如下：1型癫痫，静卧不动；2型癫痫，点头，身体偶尔抽动，尾僵直；3型癫痫，单前肢抬起，后肢痉挛；4型癫痫，双前肢抬起，站立抽动；5型癫痫，站立倒下，倒地剧烈抽动，大幅度跳跃，死亡。
(2)实验结果：
与空白对照组相比，原料大麻二酚组与除处方7外的各大麻二酚组合物处方组第一次发生强直性阵挛的时刻显著延后(P＜0.01，P＜0.05)。原料大麻二酚组小鼠的强直性阵挛总持续时间、发作频率、发作等级与空白对照组比无明显差异，各大麻二酚组合物组在这三个指标上与空白对照组相比均有改善，其中处方1-6，8可显著缩短模型小鼠强直性阵挛的总持续时间和发作频率、降低癫痫发作等级(P＜0.01，P＜0.05)。各项数据显示，与原料大麻二酚相比，本申请的大麻二酚组合物可更有效的降低癫痫严重程度，延缓癫痫发作，尤其是更低剂量的大麻二酚组合物(泊洛沙姆：CBD配比大于5：1的处方1-6和8)。
表12 不同大麻二酚制剂对戊四唑诱导的癫痫小鼠的改善作用
组别 强直性阵挛发作时间(S) 强直性阵挛持续时间(S) 强直性阵挛发作次数 平均发作等级
空白对照组 242.6±57.8 220.1±79.4 1.9±0.6 3.4±0.5
阳性对照组 373.2±69.6 ## 60.4±39.7 # 1.2±0.6 # 2.3±0.5 ##
处方1组 390.4±70.3 ## 55.9±37.8 # 1.3±0.8 # 2.4±0.3 ##
处方2组 399.6±71.1 ## 49.3±28.9 ## 1.2±0.4 # 2.3±0.7 ##
处方3组 395.7±62.6 ## 57.2±26.8 # 1.2±0.5 # 2.2±0.5 ##
处方4组 404.1±45.8 ## 45.3±30.1 ## 1.3±0.4 # 2.3±0.6 ##
处方5组 387.2±64.6 ## 52.8±34.3 ## 1.4±0.7 # 2.5±0.6 ##
处方6组 346.5±58.1 # 79.4±56.1 # 1.5±0.7 # 2.7±0.3 #
处方7组 294.5±47.1 148.7±53.1 1.7±0.6 3.1±0.6
处方8组 379.1±46.5 ## 53.7±32.7 ## 1.3±0.5 # 2.4±0.6 ##
处方9组 330.9±51.7 # 103.4±49.5 1.9±0.3 2.8±0.3
原料大麻二酚组 324.1±40.2 # 113.7±63.2 1.7±0.5 3.0±0.5
#与空白对照组相比p＜0.05，##与空白对照组相比p＜0.01
4、考察原料大麻二酚及不同大麻二酚组合物处方对强迫游泳实验中抑郁小鼠的治疗效果。
(1)实验方法
120只昆明小鼠，雌雄各半，购自北京维通利华实验动物技术有限公司。自然光12h光照/12h黑暗条件下饲养，自由饮水、摄食。
大麻二酚原料以生理盐水溶解为7.5mg/ml的混悬液供实验用；各处方大麻二酚组合物均以生理盐水溶解为5mg/ml(以CBD含量计)的水溶液或混悬液供实验用；阿米替林以生理盐水溶解为1mg/ml的水溶液或混悬液供实验用。
小鼠适应性饲养一周后，禁食12-16小时，食水自由，根据体重随机分为12组：1)空白对照组灌胃给予生理盐水；2)原料大麻二酚组灌胃给予原料大麻二酚混悬液(150mg/kg)；3)处方1-9组分别灌胃给予处方1-9的水溶液(100mg/kg)；4)阳性对照组灌胃给予阿米替林溶液(20mg/kg)。
正式测试前24小时，将小鼠置于水深10cm的玻璃圆缸(高25cm，直径10cm)内，水温24±1℃，作强迫游泳训练15分钟。将各组给药后再次将小鼠置于水深10cm的玻璃圆缸内强制游泳6min，观察并记录最后4分钟内小鼠的不动时间。当小鼠停止挣扎，浮在水中保持不动，或仅做一些必要的轻微动作保持头部浮在水面上的时间视为游泳不动时间；其中，各大麻二酚组均为在给药大麻二酚2h后再将小鼠置于玻璃圆缸内强制游泳6min，阿米替林组在给药后1h再将小鼠置于玻璃圆缸内强制游泳6min。
(2)实验结果
在小鼠的强迫游泳实验中，阿米替林、原料大麻二酚、各大麻二酚组合处方均分别产生抗抑郁作用。与原料大麻二酚组相比，除处方7外的各大麻二酚组合物处方组小鼠的不动时间进一步降低(P＜0.05)，说明更低剂量的大麻二酚组合物(泊洛沙姆：CBD配比大于5：1的处方1-8和9)比较高剂量的原料大麻二酚有更佳的抗抑郁作用。
表13 不同大麻二酚制剂对强迫游泳实验中小鼠不动时间的作用
组别 游泳不动时间(s)
空白对照组 170.4±7.1
阳性对照组 123.2±6.3 #
处方1组 94.8±6.7 ##*
处方2组 83.9±4.1 ##*
处方3组 82.5±5.4 ##*
处方4组 86.1±3.9 ##*
处方5组 91.3±5.8 ##*
处方6组 97.8±7.5 ##*
处方7组 117.2±7.9 #
处方8组 93.7±4.3 ##*
处方9组 102.1±4.7 #
原料大麻二酚组 111.5±7.7 #
#与空白对照组相比p＜0.05，##与空白对照组相比p＜0.01；*与原料大麻二酚组相比p＜0.05
5、考察原料大麻二酚及不同大麻二酚组合物处方的血浆蛋白结合率
采用常用的平衡透析法评价原料大麻二酚和本申请所述大麻二酚组合物处方1-9的血浆蛋白结合率。
(1)实验方法：
1)样品母液配置：取原料大麻二酚或大麻二酚组合物，加入甲醇配置成(200,100,50,20,10,5)μg/mL共计6个梯度，以200μg/mL作为高浓度样品，100μg/mL作为中浓度样品，50μg/mL作为低浓度样品。
2)空白透析液配置：准确称取14.110g K 2HPO 4,2.592g KH 2PO 4，1.991g NaCl溶于950ml去离子水，以0.1mol/L的HCl调pH至7.4，然后定容至1000ml。
3)透析内液：移液枪取新鲜兔血浆200μL，加入样品(大麻二酚或大麻二酚组合物母液)600μL，即为透析膜内试液。透析膜截留分子量为5000D(型号：G-RC-18-5K)，透析池为盛有20ml空白透析液的安瓿瓶。
4)专属性考察：考察血浆(或透析液)，血浆(或透析液)+样品在透析池透析平衡后，内外液的出峰情况，观察对测定结果是否存在干扰。
5)标准曲线制备：取样品母液(5～100μg/mL)各400μL，加入透析液(或血浆) 200μL，混匀后，加入400μL甲醇，离心取上清，送HPLC检验，以样品浓度为横坐标，峰面积为纵坐标，制备透析外液(或内液)的标准曲线。
6)加样回收率考察：移液枪取新鲜兔血浆200μL，分别加入高，中，低浓度样品，按5)中所述步骤，测定大麻二酚的回收率。
7)血浆蛋白结合率测定：将透析袋以去离子水洗涤后，一端结扎，分别加入高，中，低浓度的三种含药血浆(200μL+600μL)，平衡结束后，参照标曲中后处理方法进行，取样，送HPLC测定。
HPLC测定条件:
以十八烷基硅烷键合硅胶为填充剂(C18，4.6×150mm，3um色谱柱)，以乙腈-水(70:30)为流动相，检测波长为210nm，流速为1ml/min，柱温为25℃。理论板数按CBD峰计算应不低于2000。
(2)实验结果：
结果显示，除处方7外，与原料大麻二酚相比，其余各组方的血浆蛋白结合率均有明显降低。提示本申请的组合物有助于解决大麻二酚代谢中潜在的蓄积问题。
表14 不同大麻二酚制剂的血浆蛋白结合率测定结果
组别 血浆蛋白结合率％
原料大麻二酚组 95
处方1 79
处方2 74
处方3 75
处方4 72
处方5 78
处方6 83
处方7 91
处方8 81
处方9 89
尽管本发明的具体实施方式已经得到详细的描述，但本领域技术人员将理解：根据已经公开的所有教导，可以对细节进行各种修改和变动，并且这些改变均在本发明的保护范围之内。本发明的全部范围由所附权利要求及其任何等同物给出。

Claims (16)

  1. 大麻二酚组合物，其含有大麻二酚和表面活性剂；
    优选地，所述表面活性剂选自非离子型表面活性剂；例如选自聚丙二醇和/或聚氧乙烯为母体的加成物以及多元醇型的酯类或酰胺加成物；优选为泊洛沙姆；更优选地，所述泊洛沙姆分子量为9000～20000Da，例如9000～16000Da、9000～12600Da、9900～12600Da或10000～15000Da，优选9950～12600Da)，并且聚氧乙烯嵌段百分比为40％～80％,例如60％～80％、60％～75％、62％～72％、65％～75％或65％～80％；更优选地，所述泊洛沙姆为P407或分子量为9950～12600Da并且聚氧乙烯嵌段百分比为70％的泊洛沙姆；
    优选地，所述大麻二酚和表面活性剂的质量比为1：(1-100)，更优选比例为1：(5-50)。
  2. 权利要求1的组合物，其还含有抗氧化剂；
    优选地，所述抗氧化剂为有机酸或其药学上可接受的盐；
    优选地，所述抗氧化剂选自柠檬酸、酒石酸、苹果酸、琥珀酸、抗坏血酸及其药学上可接受的盐(例如钠盐或钾盐)；
    优选地，所述抗氧化剂为柠檬酸或酒石酸；优选地，所述大麻二酚和抗氧化剂的重量比为1:(0.01-0.2)；例如1:(0.01-0.1)；
    优选地，所述大麻二酚、表面活性剂和抗氧化剂的重量比为1:(1-100):(0.01-0.2)；优选为1:(5-50):(0.01-0.1)；优选为1：50：0.01。
  3. 权利要求1或2的组合物，其进一步含有药学上可接受的载体或赋形剂。
  4. 权利要求1-3任一项的组合物，其还含有所述组合物中固形物5-20倍重量的水；优选10-15倍；更优选10倍；
    优选地，所述组合物为纳米乳制剂；
    优选地，所述纳米乳的粒径为1-200nm，优选为1-100nm。
  5. 权利要求1-4任一项的组合物，其为片剂、颗粒剂、胶囊、软胶囊或注射剂。
  6. 权利要求1-5任一项的组合物在制备用于预防和/或治疗痉挛、焦虑/抑郁、炎症(例如风湿性关节炎)、癌症、多发性硬化症、癫痫特别是难治性癫痫、病毒感染或疼痛的药物中的用途。
  7. 权利要求1-5任一项的组合物在制备增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤的食品(尤其是保健食品)中的用途。
  8. 权利要求1-5任一项的组合物在以非治疗目的地祛痤疮或改善皮肤水分/油分平衡中的用途。
  9. 一种化妆品，其含有权利要求1-5任一项的组合物。
  10. 一种食品(例如保健食品)，其含有权利要求1-5任一项的组合物。
  11. 一种预防和/或治疗痉挛、焦虑/抑郁、炎症(例如风湿性关节炎)、癌症、多发性硬化症、癫痫特别是难治性癫痫、病毒感染或疼痛的方法，其包括向有此需要的受试者施用有效量的权利要求1-5任一项的组合物的步骤。
  12. 一种增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤的方法，其包括向有此需要的受试者施用有效量的权利要求1-5任一项的组合物的步骤。
  13. 一种祛痤疮或改善皮肤水分/油分平衡的方法，其包括向有此需要的受试者施用有效量的权利要求1-5任一项的组合物的步骤。
  14. 权利要求1-5任一项的组合物，其用于预防和/或治疗痉挛、焦虑/抑郁、炎症(例如风湿性关节炎)、癌症、多发性硬化症、癫痫特别是难治性癫痫、病毒感染或疼痛。
  15. 权利要求1-5任一项的组合物，其用于增强免疫力、辅助降血糖/血脂、辅助改善记忆、清咽、改善睡眠、减肥或改善化学性肝损伤。
  16. 权利要求1-5任一项的组合物，其用于祛痤疮或改善皮肤水分/油分平衡。
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