Δ 8 -Synthesis method of isoTHC and Δ 8 - Applications of different THC

Abstract

The application discloses a kind of Δ 8 - Synthesis method and Δ 8 - Application of THC. Wherein, the synthesis method comprises the following steps: taking cannabidiol as raw material, adding organic solvent, converting under ultraviolet light to generate Δ 8 - THC. According to the technical scheme of the application, cannabidiol CBD is converted into Δ 8 - THC under the catalysis of ultraviolet light, so that Δ 8 - THC appears as main product, and the method for converting CBD also ingeniously avoids complex and high-cost total synthesis route.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically, to a Δ 8 -Synthesis method of isoTHC and Δ 8 - Applications of different THC. Background Technology

[0002] Cannabis sativa, also known as mountain hemp, sesame, wild hemp, or fire hemp, is an annual herbaceous plant belonging to the Moraceae family. The seeds of cannabis are often used medicinally, known as "fire hemp seeds." Modern pharmacological research shows that cannabis possesses analgesic, intraocular pressure-lowering, antitumor, antiemetic, sedative, antithrombotic, antibacterial, and anti-inflammatory biological activities. In modern clinical treatment, cannabis has shown significant therapeutic effects on chronic pain, glaucoma, malignant gliomas, atherosclerosis, AIDS-related weight loss syndrome, and adverse reactions after cancer chemotherapy and radiotherapy. The United Nations listed cannabis as one of the three major narcotic drugs alongside heroin and cocaine in the 1961 Single Convention on Narcotic Drugs and the 1971 Convention on Psychotropic Substances and its amendments. Global control over cannabis has been relatively strict, leading to slow progress in cannabis research. However, with the World Health Organization recognizing cannabidiol (CBD), the inactive addictive component of cannabis, and including it on the international list of uncontrolled substances, international policies on industrial hemp are gradually being relaxed.

[0003] Based on their THC (cannabidiol) content, cannabis can be divided into two categories: industrial hemp (THC < 0.3%, showing no psychoactive effects) and recreational cannabis (THC > 0.5%, exhibiting significant psychoactive effects and a tendency for abuse). Cannabidiol (CBD) is an important compound extracted from industrial hemp. When we talk about industrial hemp, we are usually referring to CBD. It is unrelated to the efficacy of drugs affecting mental state and does not affect motor function, memory, or body temperature. Due to its good safety profile and lack of psychoactive effects, it can be used to treat conditions including inflammation, neurodegenerative diseases, epilepsy, autoimmune diseases such as multiple sclerosis, arthritis, schizophrenia, and cancer.

[0004] Cannabis, a plant in the Moraceae family, contains secondary metabolites of hundreds of chemical entities, each with potentially interesting biological properties. More than 90 cannabinoids are known, and besides THC and CBD, other typical cannabinoids from plant sources include cannabinoids (CBC), cannabinol (CBG), cannabidiol (CBDV), and tetrahydrocannabinol (THCV). Although these compounds have similar chemical structures, they can induce different pharmacological effects, indicating significant potential for development.

[0005] Transient receptor potential (TRP) channels are involved in a wide range of physiological functions, from the transmission of vision, hearing, and taste, to nociception and temperature sensation, and the control of muscle contraction and vasomotor activity. Furthermore, TRP channels are key players in regulating intestinal motility, mineral absorption, blood circulation, bladder and airway hypersensitivity, fluid homeostasis, and cell growth and survival. This makes the TRP family important therapeutic targets for many major diseases, such as cardiovascular disease, neurodegenerative diseases, pain, and cancer. There are 6 subfamilies and 28 members of TRP channels in mammals, among which TRPV1, TRPA1, and TRPM8 are currently the focus of research.

[0006] Transient receptor potential channel protein (TRPA1), a cold-sensitive ion channel in the TRP family, has been found to be expressed in blood vessels, intestines, pancreas, and myocardial tissue in recent years. Activation of TRPA1 regulates the secretion of gastrointestinal hormones such as GLP-1, ghrelin, 5-HT, and cholecystokinin, inhibiting gastric emptying and thus affecting food intake, which may be an important mechanism for its weight-loss effect. Activation of pancreatic β-channel TRPA1 can promote insulin secretion, and its ability to promote GLP-1 secretion may also contribute to regulating glycemic homeostasis. Activation of TRPA1 in endothelial cells can improve endothelial-dependent vasodilation by promoting NO release, including neurogenic and smooth muscle mechanisms. Furthermore, as a cold channel, low temperature can significantly prolong lifespan by activating TRPA1.

[0007] The TRPM8 channel is also a cold-activated channel, primarily expressed in somatosensory neurons of the dorsal root ganglion and trigeminal ganglion. It can be activated by various chemical and physical stimuli, such as menthol, phosphatidylinositol-4,5-bisphosphate (PIP2), and voltage, thereby activating cellular signaling to participate in a wide range of physiological processes. Currently, TRPM8 has been found to play a crucial role as a major mediator of menthol-induced acute and migraine headaches, making it a potential therapeutic target for cold-related pain, chronic pain, and migraines. Furthermore, the TRPM8 channel is also involved in processes such as irritable bowel syndrome, oropharyngeal dysphagia (OD), and chronic cough; therefore, the TRPM8 receptor is considered a valuable therapeutic target for the development of related targeted drugs. In addition, there are numerous research reports on TRPM8 channels as targets in cancer treatment, showing promising therapeutic effects in prostate cancer, breast cancer, colon cancer, lung cancer, and skin cancer.

[0008] WO02070506 reported on the synthesis of Δ using CBD as a raw material. 8 -THC (compound 1) and Δ 9 The method of -THC(compound 2), wherein byproducts include Δ 8 -Irrelevant THC, patent report on Δ9 -THC is the main component of cannabis, Δ 8 -THC ratio Δ 9 -THC exhibits better stability, lower psychoactive activity, stronger antiemetic activity, and no Δ 8 - Reports on the activity of isoTHC.

[0009]

[0010] Journal of the American Chemical Society 1974, 96(18): 5860, Collect.Czech.Chem.Commun.(Vol.66)(2001): 1257, GB2567235 reported a method for the chemical synthesis of THC, which requires the use of oxidants such as ozone and has a low yield. It mentions the byproduct Δ 8 -IsoTHC, but no reports of pharmacological activity.

[0011] Existing technology has only reported Δ 8 -The chemical pathway for the acquisition of isoTHC, in which Δ 8 -IsoTHC occurs as a byproduct, with a very low yield of only about 10%.

[0012] In summary, there is no Δ in the existing technology. 8 - Any reports of pharmacological activity of isoTHC. Summary of the Invention

[0013] This invention aims to provide a Δ 8 -Synthesis method of isoTHC and Δ 8 - Applications of different THC to explore Δ 8 - Preparation and pharmaceutical applications of isoTHC.

[0014] To achieve the above objectives, according to one aspect of the present invention, a Δ 8 A method for synthesizing -isoTHC. This method includes the following steps: using cannabidiol as a raw material, adding an organic solvent, and converting it under ultraviolet light to generate Δ... 8 - Different THC.

[0015] Further, after the conversion is complete, the product is concentrated to dryness under reduced pressure to obtain the crude product Δ. 8 - Different THC.

[0016] Furthermore, the crude product Δ 8 -IsoTHC was purified by silica gel normal-phase column chromatography, followed by purification by C18 silica gel reverse-phase column chromatography to obtain Δ 8 - Different THC.

[0017] Furthermore, after the cannabidiol is added to an organic alcohol solvent, a dissolution step is included before conversion under ultraviolet light.

[0018] Further, the organic solvent is one or more selected from the group consisting of methanol, ethanol, isopropanol, acetonitrile, methyl acetate, ethyl acetate, tetrahydrofuran, dichloromethane or chloroform; preferably, the organic solvent is ethanol, methanol or acetonitrile.

[0019] Furthermore, the wavelength of the ultraviolet light is 200–400 nm; preferably, the wavelength of the ultraviolet light is 254 nm.

[0020] Furthermore, the conversion time is 2 to 14 days, preferably 10 days.

[0021] According to another aspect of the present invention, a drug is provided that activates the TRPA1 channel and inhibits the TRPMI channel. The drug comprises an effective dose of Δ 8 - Different THC.

[0022] According to another aspect of the invention, Δ is provided 8 - The use of isoTHC in the preparation of medicaments for treating diseases that benefit from TRPA1 channel activation, preferably diseases that benefit from TRPA1 channel activation including obesity, high blood pressure or high blood sugar.

[0023] According to another aspect of the invention, Δ is provided 8 - The use of isoTHC in the preparation of medicaments for treating diseases that benefit from TRPMI channel inhibition, preferably, the diseases that benefit from TRPMI channel inhibition include any one of chronic pain, migraine, irritable bowel syndrome, dysphagia, chronic cough and cancer; more preferably, the cancers include any one of prostate cancer, breast cancer, colon cancer, lung cancer and skin cancer.

[0024] Using the technical solution of this invention, cannabidiol (CBD) is converted into Δ under ultraviolet light catalysis. 8 - Different THC, making Δ 8 -IsoTHC appears as the main product, and the CBD conversion method cleverly avoids the complex and costly total synthesis route. Attached Figure Description

[0025] The accompanying drawings, which form part of this application, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings:

[0026] Figure 1 Δ is shown in Example 1 8 -Mass spectrum of isoTHC;

[0027] Figure 2 Δ is shown in Example 1 8 - The hydrogen NMR spectrum of different THC. Detailed Implementation

[0028] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0029] The inventors of this invention made a surprising discovery that cannabidiol (CBD) can be converted into Δ in one step under ultraviolet light irradiation. 8 -IsoTHC, compared with total synthesis methods, this method is simpler to operate, relatively lower in cost, and more environmentally friendly. Furthermore, because in existing reports, Δ 8 -Iso-THC are all byproducts, therefore this method fills the gap in Δ 8 - There is no chemical synthesis method for isoTHC.

[0030] Existing literature on Δ 8 Reports on -isoTHC are scarce, and there are currently no reports on its activity. However, due to Δ 8 -IsoTHC has a structure very similar to that of natural cannabinoids, and is similar to the addictive component Δ 9 -THC has a different structure, therefore Δ 8 -IsoTHC has great potential for pharmaceutical development. This invention investigated its activity and found that Δ 8 -Isothionein THC exhibits activity against TRPA1 and TRPM8 channels, potentially leading to Δ 8 -IsoTHC plays a role in weight loss, lowering blood pressure, lowering blood sugar, anti-aging, and treating pain, chronic cough, and various cancers. Additionally, Δ 8 -IsoTHC has almost no effect on the TRPV1 channel, so it also exhibits a certain degree of channel selectivity when it is formulated into a drug, which helps to avoid adverse reactions.

[0031] According to a typical embodiment of the present invention, a Δ 8 A method for synthesizing -isoTHC. This method includes the following steps: using cannabidiol as a raw material, adding an organic solvent, and converting it under ultraviolet light to generate Δ... 8 -IsoTHC. Cannabidiol (CBD) can be converted into ΔTHC in one step under ultraviolet light irradiation. 8 Compared with total synthesis methods, this method for -isoTHC is simpler to operate, relatively lower in cost, and more environmentally friendly.

[0032] Preferably, after conversion, the product is concentrated to dryness under reduced pressure to obtain crude product Δ. 8- IsoTHC. The temperature for vacuum concentration is related to the boiling point of the solvent. If the temperature is too low, the solvent will not evaporate completely; if the temperature is too high, it is prone to boiling over. Therefore, the temperature and specific parameters for vacuum concentration can be determined by those skilled in the art based on the specific circumstances.

[0033] According to a typical embodiment of the present invention, the crude product Δ 8 -IsoTHC was purified by silica gel normal-phase column chromatography, followed by purification by C18 silica gel reverse-phase column chromatography to obtain Δ 8 - Different THC.

[0034] According to a typical embodiment of the present invention, after cannabidiol is added to an organic alcohol solvent and before it is converted under ultraviolet light, a step of dissolving (completely dissolving) at room temperature is also included.

[0035] Preferably, the organic solvent is one or more selected from the group consisting of methanol, ethanol, isopropanol, acetonitrile, methyl acetate, ethyl acetate, tetrahydrofuran, dichloromethane, or chloroform; preferably, the organic solvent is ethanol, methanol, or acetonitrile.

[0036] According to a typical embodiment of the present invention, the wavelength of the ultraviolet light is 200-400 nm; preferably, the wavelength of the ultraviolet light is 254 nm.

[0037] According to a typical embodiment of the present invention, the conversion time is 2 to 14 days, preferably 10 days.

[0038] According to a typical embodiment of the present invention, a drug is provided that activates the TRPA1 channel and inhibits the TRPMI channel. The drug comprises an effective dose of Δ 8 -IsoTHC. This drug can be administered orally (including capsules, tablets, oral solutions, etc.), injected (including intravenous injections, small-volume injections), or transdermal.

[0039] According to a typical embodiment of the present invention, Δ is provided 8 - The use of isoTHC in the preparation of medicaments for treating diseases that benefit from TRPA1 channel activation, preferably diseases that benefit from TRPA1 channel activation including obesity, high blood pressure or high blood sugar.

[0040] According to a typical embodiment of the present invention, Δ is provided 8 - The use of isoTHC in the preparation of medicaments for treating diseases that benefit from TRPMI channel inhibition, preferably, the diseases that benefit from TRPMI channel inhibition include any one of chronic pain, migraine, irritable bowel syndrome, dysphagia, chronic cough and cancer; more preferably, the cancers include any one of prostate cancer, breast cancer, colon cancer, lung cancer and skin cancer.

[0041] The beneficial effects of the present invention will be further illustrated below with reference to embodiments. Reagents or experimental procedures not described in detail in this invention can be implemented using conventional reagents or experimental procedures in the art by those skilled in the art, without affecting the final results of the present invention.

[0042] Example 1

[0043]

[0044] 5g of cannabidiol (CBD) was dissolved in 5L of acetonitrile and irradiated under ultraviolet light (wavelength: 254nm) for 10 days. After the solvent was concentrated under reduced pressure, it was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). Then it was purified by C18 reversed-phase silica gel column separation (acetonitrile / water = 55:45-70:30). After the solvent was concentrated under reduced pressure, about 3.2g of product, namely Δ8-isoTHC, was obtained.

[0045] Ms: 314, [M+H] + =315.

[0046] 1 H-NMR (600MHz, CDCl3), δ6.28-6.12ppm (s, 2H), δ4.99-4.92ppm (d, 2H), δ4.56ppm (s, 1H), δ3.47-3.46ppm (d, 1H), δ2.47-2. 44ppm (t, 2H), δ2.34ppm (s, 1H), δ1.88-1.86ppm (d, 4H), δ1.77~1.52 (m, 7H), δ1.34~1.25ppm (m, 7H), δ0.90~0.88ppm (t, 3H).

[0047] Δ 8 -Mass spectra of isoTHC are attached. Figure 1 The proton NMR spectrum is attached. Figure 2 .

[0048] Example 2

[0049] 10g of cannabidiol (CBD) was dissolved in 10L of acetonitrile and irradiated under ultraviolet light (wavelength: 365nm) for 12 days. After concentrating the solvent under reduced pressure, the solution was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). It was then purified again by C18 reversed-phase silica gel column separation (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 6.8g of the product was obtained, i.e., Δ 8 - Different THC.

[0050] Example 3

[0051] 10g of cannabidiol (CBD) was dissolved in 5L of tetrahydrofuran and irradiated under ultraviolet light (wavelength: 254nm) for 14 days. After concentrating the solvent under reduced pressure, the product was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). It was then purified again by C18 reversed-phase silica gel column separation (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 2.8g of the product was obtained, i.e., Δ 8 - Different THC.

[0052] Example 4

[0053] 10g of cannabidiol (CBD) was dissolved in 10L of methanol and irradiated under ultraviolet light (wavelength: 254nm) for 2 days. After concentrating the solvent under reduced pressure, the product was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). It was then purified again by C18 reversed-phase silica gel column separation (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 3.4g of the product was obtained, i.e., Δ 8 - Different THC.

[0054] Example 5

[0055] 10g of cannabidiol (CBD) was dissolved in 10L of ethyl acetate and irradiated under ultraviolet light (wavelength: 365nm) for 12 days. After concentrating the solvent under reduced pressure, the product was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). It was then purified again by C18 reversed-phase silica gel column chromatography (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 2.1g of the product was obtained, i.e., Δ 8 - Different THC.

[0056] Example 6

[0057] 10g of cannabidiol (CBD) was dissolved in 10L of a mixed solvent of chloroform and methanol (v / v = 1:1) and irradiated under ultraviolet light (wavelength: 254nm) for 14 days. After concentrating the solvent under reduced pressure, the product was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1); then purified by C18 reversed-phase silica gel column separation (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 1.6g of product was obtained, i.e., Δ 8 - Different THC.

[0058] Example 7

[0059] 10g of cannabidiol (CBD) was dissolved in 10L of isopropyl alcohol and irradiated under ultraviolet light (wavelength: 365nm) for 7 days. After concentrating the solvent under reduced pressure, the product was purified twice by silica gel column chromatography (200-300 mesh silica gel, PE:EA = 150:1-80:1). It was then purified again by C18 reversed-phase silica gel column chromatography (acetonitrile / water = 55:45-70:30). After concentrating the solvent under reduced pressure, approximately 4.1g of the product was obtained, i.e., Δ 8 - Different THC.

[0060] Test materials

[0061] Test sample: Δ 8 - Iso-THC, self-made, purity 98.45%.

[0062] Positive controls: cinnamaldehyde (98% purity), ruthenium red (approx. 100% purity), capsaicin (approx. 100% purity), capsicum (99.3% purity), menthol (97% purity), 2-APB (approx. 100% purity).

[0063] Patch clamp detection Δ 8 - Effects of isoTHC on TRPV1, TRPA1, and TRPM8

[0064] TRPA1

[0065] The voltage stimulation protocol for whole-cell patch-clamp recording of TRPA1 currents was as follows: After whole-cell occlusion, the cell membrane voltage was clamped at -80 mV. The recording voltage was stepped from 0 mV to -100 mV and held for 1 ms, followed by a 300 ms ramp stimulation to 100 mV. Data was collected every 5 s to observe the effect of the drug on TRPA1 currents. Experimental data were acquired using an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0066] TRPV1

[0067] The voltage stimulation protocol for whole-cell patch-clamp recording of TRPV1 currents was as follows: After whole-cell occlusion, the cell membrane voltage was clamped at 0 mV. The recording voltage was stepped from 0 mV to -100 mV and held for 1 ms, followed by a 300 ms ramp stimulation to 100 mV. Data was collected every 5 s to observe the effect of the drug on TRPV1 currents. Experimental data were acquired using an EPC-10 amplifier (HEKA Electronics) and stored in PatchMaster (HEKA Electronics) software.

[0068] TRPM8

[0069] The voltage stimulation protocol for whole-cell patch-clamp recording of TRPM8 currents was as follows: After whole-cell occlusion, the cell membrane voltage was clamped at -80 mV. The recording voltage was stepped from 0 mV to -100 mV, followed by a 500 ms ramp stimulation to 100 mV. Data was collected repeatedly every 5 s to observe the effect of the drug on TRPM8 currents. Experimental data were acquired using an EPC-10 amplifier (HEKA Electronics) and stored in PatchMaster (HEKA Electronics) software.

[0070] A capillary glass tube was drawn into a recording electrode using a microelectrode drawing instrument. Under an inverted microscope, the microelectrode manipulator was used to bring the recording electrode into contact with the cell, and negative pressure was applied to aspirate and form a GΩ seal. After forming the GΩ seal, rapid capacitance compensation was performed, and then negative pressure was continued to rupture the cell membrane, establishing a whole-cell recording mode. Slow capacitance compensation was then performed, and the membrane capacitance and series resistance were recorded. No leakage compensation was applied.

[0071] A coverslip containing cells was placed in the recording bath of an inverted microscope. The working solution containing the test compound and the external solution without the compound were perfused through the recording bath by gravity to act on the cells. A vacuum pump was used for fluid exchange during recording. Three cells were independently and repeatedly tested. All electrophysiological experiments were performed at room temperature.

[0072] Δ was detected in three independent repeated trials. 8 The agonistic and inhibitory effects of isoTHC on TRPA1, TRPV1, and TRPM8 channels are summarized in Table 1-3 below:

[0073] Table 1 Δ 8 - Results of the effects of isoTHC on TRPA1 current and the positive control ruthenium red on TRPA1 current:

[0074]

[0075] The result is Δ 8 -Irrelevant THC has an excitatory effect on the TRPA1 channel.

[0076] Table 2 Δ 8 - Results of the inhibitory effects of isoTHC on TRPV1 current and the positive control capsazepine on TRPV1 current:

[0077]

[0078] The result is Δ 8 -Iso-THC showed almost no inhibitory effect compared to the positive control.

[0079] Table 3 Δ 8- Results of the inhibitory effect of isoTHC on TRPM8 current and the inhibitory effect of positive control 2-APB on TRPM8 current:

[0080]

[0081] The result is Δ 8 -Iso-THC showed a stronger inhibitory effect compared to the positive control, completely blocking the TRPM8 ion channel.

[0082] From the above description, it can be seen that the above embodiments of the present invention achieve the following technical effects: The present invention has studied its activity and found that Δ 8 -Isothionein THC exhibits activity against TRPA1 and TRPM8 channels, potentially leading to Δ 8 -IsoTHC plays a role in weight loss, lowering blood pressure, lowering blood sugar, anti-aging, and treating pain, chronic cough, and various cancers. Additionally, Δ 8 -IsoTHC has almost no effect on the TRPV1 channel, so it also exhibits a certain degree of channel selectivity when it is formulated into a drug, which helps to avoid adverse reactions.

[0083] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A kind of Δ 8 The method for synthesizing -isoTHC is characterized by, Includes the following steps: Using cannabidiol as a raw material, an organic solvent is added, and the mixture is transformed under ultraviolet light to produce Δ 8 - IsoTHC; the organic solvent is selected from methanol, ethanol, isopropanol, and acetonitrile; the conversion time is 2 to 14 days; the wavelength of the ultraviolet light is 200 to 400 nm.

2. The synthesis method according to claim 1, characterized in that, After the conversion is completed, the product is concentrated to dryness under reduced pressure to obtain crude product Δ. 8 - Different THC.

3. The synthesis method according to claim 2, characterized in that, The crude product Δ 8 -IsoTHC was purified by silica gel normal-phase column chromatography, followed by purification by C18 silica gel reverse-phase column chromatography to obtain Δ 8 - Different THC.

4. The synthesis method according to any one of claims 1 to 3, characterized in that, The process includes a dissolution step at room temperature after the cannabidiol is added to an organic alcohol solvent and before conversion under ultraviolet light.

5. The synthesis method according to claim 1, characterized in that, The organic solvent is ethanol, methanol, or acetonitrile.

6. The synthesis method according to claim 1, characterized in that, The wavelength of the ultraviolet light is 254 nm.

7. The synthesis method according to claim 1, characterized in that, The conversion process takes 10 days.

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