Preparation for and use of cannabidiol derivative drugs

Abstract

CBD derivatives effective for treating opioid addiction, neurodegenerative diseases and neuroinflammation. The CBD derivatives have lower toxicity than CBD, are not addictive, and do not inhibit respiration or damage short-time memory.

Description

Preparation and application of cannabidiol derivative drugs Technical Field

[0001] The present invention relates to cannabidiol (CBD) derivatives for treating neurodegenerative diseases and opioid addiction, and more particularly, to CBD analogs having anti-neuroinflammatory and pyruvate kinase M2 (PKM2) regulating activities for treating neurodegenerative diseases and opioid addiction. Background Art

[0002] Opioids refer to morphine extracted from the opium poppy and its synthetic derivatives. Opioid addiction is a chronic, relapsing disease that poses a serious public health problem. Currently, there is a lack of effective treatments and methods to address opioid addiction. According to the World Health Organization, opioids cause 69,000 deaths each year. An estimated 15 million people are dependent on opioids. Traditional treatments for opioid addiction include medication maintenance therapy, withdrawal therapy, comprehensive detoxification, and physical therapy. However, these treatments have significant limitations.

[0003] Maintenance drug therapy typically achieves detoxification through the administration of weak opioid receptor agonists, such as methadone or methylphenidate, μ-opioid receptor (MOR) antagonists, such as naltrexone or naloxone, or gradually reduced doses of oral opioid solutions. However, such treatments often exhibit limited efficacy and significant side effects. Furthermore, weak opioid receptor agonists themselves are addictive, making complete recovery difficult for many opioid-addicted patients. It has now been well documented that morphine causes neuroinflammation, releases proinflammatory cytokines, regulates neuroexcitability in the central nervous system associated with drug abuse, and influences morphine dependence and reward.

[0004] Treatment for withdrawal symptoms typically involves a combination of withdrawal induction and management of withdrawal symptoms. Commonly used medications include promethazine (an α2-agonist), which effectively suppresses symptoms such as vomiting, diarrhea, hypertension, and rapid breathing. However, these medications often also cause adverse effects, such as anxiety and insomnia, and are therefore often insufficient to maintain abstinence from opioids.

[0005] Comprehensive detoxification methods often use scopolamine (an M-cholinergic receptor blocker) to antagonize the vagus nerve, control withdrawal symptoms caused by morphine addiction, reduce or reverse morphine tolerance, and promote drug excretion. However, this drug only inhibits the physical dependence caused by morphine, namely the withdrawal symptoms. Therefore, comprehensive detoxification methods usually have little effect on psychological dependence.

[0006] Physical therapy for opioid addiction treatment generally includes exercise, acupuncture, and other physical therapies, but is generally limited in its effectiveness in treating opioid addiction.

[0007] Therefore, there is an urgent need for highly effective and low-side effect treatments for physical and psychological dependence on opioids.

[0008] Summary of the Invention

[0009] This patent relates to the groundbreaking discovery of a class of CBD derivatives that have lower toxicity and side effects and exhibit anti-neuroinflammatory activity similar to CBD. Although CBD exhibits anti-neuroinflammatory activity, the compound also has considerable neurotoxic effects, which limits the scope of its therapeutic applications. To address this problem, this patent adopts a function-oriented synthesis strategy to improve the therapeutic index by optimizing the CBD side chain pharmacophore. This strategy has generated many improved CBD analogs that have higher activity and lower toxicity relative to CBD. For example, an illustrative compound of the present invention, compound 19a (CIAC001), has a therapeutic index of approximately 30 times that of CBD.

[0010] Even more surprising is the potential therapeutic activity of these compounds in treating opioid addiction. Specifically, animal studies have shown that these compounds inhibit morphine-induced inflammation and microglial activation in the medial prefrontal cortex (mPFC), blocking morphine-induced withdrawal reactions, behavioral sensitization, and conditioned place preference (CPP) behavior. Furthermore, these compounds lack addictive reward effects and show no significant activity at CB1 / CB2 receptors.

[0011] The present invention provides a method for treating or preventing opioid addiction, withdrawal or overdose, comprising administering a compound according to formula I:

[0012] or a salt or solvate thereof, wherein the main body of the compound comprises:

[0013] R 1 Selected from the following structures:

[0014] -H, or

[0015] R 2A and R 2B are independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C4, wherein R 2A and R 2B You can choose diazirine instead;

[0016] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0017] Subscript n ranges from 0 to 12;

[0018] thereby treating or preventing opioid addiction, withdrawal, or overdose.

[0019] In some cases, the compound of formula I may be preferably within the scope of the compound of formula Ia:

[0020] In some cases, R 1 Selected from the following structures: -H, or In some cases, R 1 for

[0021] In some cases, R 2A and R 2B are independently selected from the following structures: -H or C1-C4 alkane. In some cases, R 2A and R 2B Each is independently selected from the following structures: -H or -CH3. In certain cases, R 2A and R 2B In some cases, n is -H. In some cases, n is 0 or 1. In some cases, n is 0.

[0022] In certain cases, the compound of formula Ia has the structure

[0023] This invention includes compounds of Formula I for use in treating or preventing opioid addiction and withdrawal symptoms.

[0024] The compound of formula I is administered at a dose of 0.5 μg / kg, 2 μg / kg, 20 μg / kg, 200 μg / kg, 2 mg / kg, 20 mg / kg or 100 mg / kg. In some cases, the C of the compound of formula I in the brain of the subject is MAX In some cases, the brain to plasma ratio of the compound of Formula I is at least about 10:1, 25:1, 50:1, 100:1 or 200:1 1 hour after administration.

[0025] The opioids referred to in this invention include fentanyl, ibuprofen, codeine, diazepam, dihydrocodeine, enkephalin, heroin, oxycodone, oxymorphone, meperidine, meprofen, methadone, morphine, nikomeidine, opium, oxycodone, oxymorphone, chenotetin, or derivatives, precursors, or pharmacologically acceptable salts or solvents thereof. In some cases, the opioid refers specifically to morphine. The subjects referred to in this invention are morphine addicts.

[0026] The subjects of this invention exhibit symptoms of opioid addiction, including withdrawal symptoms, behavioral sensitization, or conditioned place preference

[0027] In this invention, the compound of Formula I is administered orally, subbuccally, sublingually, rectally, vaginally, intravenously, intraarterially, intramedullary, intramuscularly, intracerebrally, intraventricularly, intraspinally, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, intramucosally, or via an inhaler. In some cases, the compound of Formula I is administered with a pharmacologically acceptable excipient. In some cases, the invention further comprises administering an opioid antagonist to the subject. In certain instances, the opioid antagonist is naloxone or naltrexone.

[0028] A sufficient dose of the compound of Formula I can inhibit opioid-induced overexpression of proinflammatory cytokines such as IL-1β, TNFα, IL-6, or a combination thereof in a subject. A sufficient dose of the compound of Formula I can modulate PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, or elongation factor 1γ. In certain specific cases, a sufficient dose of the compound of Formula I can modulate PKM in a subject. In certain specific cases, PKM is specifically expressed as PKM2. A sufficient dose of the compound of Formula I can increase the ratio of M2 to M1 microglia in the brain of a subject. A sufficient dose of the compound of Formula I does not induce a toxic stress response in the subject.

[0029] In an embodiment, the present invention discloses a method for regulating PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, and elongation factor 1γ in a subject, comprising administering a compound of formula I:

[0030] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures:

[0031] -H, or

[0032] R 2A and R 2Bare independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C4, wherein R 2A and R 2B You can choose to use diazirine instead;

[0033] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0034] Subscript n ranges from 0 to 12;

[0035] It thereby regulates PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, and elongation factor 1γ in the subject's body.

[0036] In some cases, the method comprises modulating a PKM in the subject. In certain cases, the PKM is PKM2.

[0037] In an embodiment, the present disclosure provides a method for increasing the ratio of M2 to M1 microglia in the brain of a subject, comprising administering to the subject a compound of formula I:

[0038] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures:

[0039] -H, or

[0040] R 2A and R 2B are independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C4, wherein R 2A and R 2B You can choose to use diazirine instead;

[0041] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0042] Subscript n ranges from 0 to 12;

[0043] This increased the ratio of M2 to M1 microglia in the subjects' brains.

[0044] In another aspect, the present invention provides a method for treating a neurodegenerative disease, comprising administering to a subject a compound of formula I:

[0045] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures:

[0046] -H, or

[0047] R 2A and R 2B are independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C4, wherein R 2A and R 2B You can choose to use diazirine instead;

[0048] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0049] Subscript n ranges from 0 to 12;

[0050] thereby treating neurodegenerative diseases.

[0051] In some cases, the compound of formula I may be preferably a compound of formula Ia:

[0052] In special cases, R 1 Selected from the following structures: H, or More preferably, R 1 for

[0053] In some cases, R 2A and R 2B Each is independently selected from the following structures: -H or C1-C4 alkane. In other cases, R 2A and R 2B are independently selected from the following structures: -H or -CH3. Or, R 2A and R 2B All are -H.

[0054] In other cases, R 3A and R 3B Each of the following structures is independently selected: -H, -Cl, -Br, or -COOH. In some other cases, R 3A and R 3B All are -H. n is 0 or 1.

[0055] In special cases, the compound structure of formula I is

[0056] In some cases, the disease or disorder is selected from Alzheimer's disease, Parkinson's disease, or amyotrophic lateral sclerosis. The compound of Formula I inhibits neuroinflammation in a subject. In certain instances, the compound of Formula I has an IC50 activity for inhibiting LPS-induced excessive nitric oxide production in microglia. 50 In a specific embodiment, the cytotoxicity of the compound of Formula I in microglia is IC 50 At least about 1 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 75 μM, 100 μM, 150 μM, or 200 μM.

[0057] Administration of a sufficient dose of a compound of Formula I can reduce the expression of IL-1β in the microglia of a subject by at least about 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold, or at least about 20-fold. On the other hand, administration of a sufficient dose of a compound of Formula I can reduce the expression of TNFα in the microglia of a subject by at least about 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold, or at least about 20-fold. Administration of a sufficient dose of a compound of Formula I can reduce the expression of IL-6 in the microglia of a subject by at least about 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold, or at least about 20-fold.

[0058] In specific aspects, the compound of Formula I at a dose of about 0.5 μg / kg, 2 μg / kg, 20 μg / kg, 200 μg / kg, 2 mg / kg, 20 mg / kg, or about 100 mg / kg can increase the C of the compound of Formula I in the brain of a subject. MAX One hour after administration to a subject, the brain-plasma drug concentration ratio of the compound of Formula I is at least about 10:1, 25:1, 50:1, 100:1, or at least about 200:1.

[0059] The compound of formula I is administered orally, subbuccal, sublingual, rectally, vaginally, intravenously, intraarterially, intraosseously, intramuscularly, intracerebrally, intraventricularly, submeningeal, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, submucosally, or by inhaler. Simultaneously, the compound of formula I can be administered together with an acceptable excipient.

[0060] Administration of a compound of Formula I is sufficient to modulate PKM, 60 kDa heat shock protein, α-enolase, eukaryotic translation elongation factor 1 α1, RNA helicase, heat shock protein 90-beta, ubiquitin carboxyl-terminal hydrolase, T-complex protein 1 subunit α, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor, clade B member 6a, or elongation factor 1-γ in a subject. In certain instances, the compound of Formula I is sufficient to modulate PKM in a subject. In certain instances, the PKM is specifically PKM2.

[0061] The administration of the compound of Formula I is sufficient to increase the ratio of M2 to M1 microglia in the brain of the subject. In another aspect, the compound of Formula I does not induce a toxic stress response in the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figures 1A-D show statistical graphs of microglial mRNA expression and morphology. Figures 1A-C show the mRNA expression of IL-1β, TNF-α, and IL-6 in BV-2 cells treated with LPS, varying amounts of CIAC001, and CBD, respectively. Figure 1D shows morphological images of microglial cells treated with LPS, LPS, and LPS and CIAC001, and also shows the percentages of rod-shaped, oval, and round microglia in each treatment group.

[0063] Figures 2A-H show images of Iba1 staining, a microglial marker, in mice subjected to a morphine addiction behavior test and mice treated with a combination of morphine, CIAC001, and CBD. Figure 2A shows the results of the morphine withdrawal jump test. Figure 2B shows the results of the behavioral sensitization test during the induction phase. Figure 2C shows a graph of the results of the behavioral sensitization test during the expression phase. Figure 2D shows a graph of the CPP scores. Figures 2E-G show images of IL-1β mRNA expression in the mPFC, nucleus accumbens (NAc), and ventral tegmental area (VTA) of mice. Figure 2H shows a panel of images of the microglial marker Iba1 in the mPFC, as well as a bar graph of the number of Iba1-positive cells per square millimeter.

[0064] Figure 3A-F shows a set of graphs for the CIAC001 safety assessment performed on mice. Figure 3A is a graph of the respiratory rate of mice treated with blank, tetrahydrocannabinol (THC) and CIAC001. Figure 3B is a graph of the time mice spent on the "new arm" in a 2-minute Y-maze test. Figure 3C is a graph of the CPP scores of mice administered with CIAC001 on the first test day and the subsequent 6-day training. Figure 3D is a graph of the motor activity of mice administered with CIAC001 on the first test day of CPP and the subsequent 6-day training. Figure 3E is a graph of the grooming and scratching frequency of mice administered with CIAC001 on the first test day of CPP and the subsequent 6-day training. Figure 3F is a graph of the weight changes of mice in CIAC001 or blank treatment for 7 consecutive days.

[0065] Figures 4A-C show a schematic diagram of a CIAC001 pull-down experiment, a silver-stained image of proteins enriched using the pull-down probe, and a Venn diagram showing proteins identified from the pull-down experiment by LC-MS / MS. Figure 4A is a schematic diagram of a pull-down experiment designed to identify proteins that bind to CIAC001. Figure 4B is a silver-stained image of enriched proteins collected from the pull-down experiment described in Figure 4A. Figure 4C is a Venn diagram summarizing the proteins collected from the experiment described in Figure 4A and identified by LC-MS / MS.

[0066] Figures 5A-F show a set of Western blot results and PKM2 molecular docking maps. Figure 5A shows the Western blot / pull-down results of PKM2 with compounds 19e and 19f. Figure 5B shows images and results from a PKM2 thermostability assay, depicting the interaction between endogenous PKM2 and CIAC001. Figure 5C shows the change in autofluorescence of PKM2 after incubation with different concentrations of CIAC001. Figure 5D shows the conformation of PKM2 bound by CIAC001 and compound 19b. Figure 5E shows the key PKM2 residues bound by CIAC001. Figure 5F shows the key PKM2 residues bound by 19b.

[0067] Figures 6A-E show a set of graphs and Western blot results of BV-2 cells treated with CIAC001 or compound 19b. Figure 6A shows ATP levels in BV-2 cells treated with vehicle, CIAC001, or compound 19b. Figure 6B shows Western blot results showing PKM2 tetramer and monomer levels in cells treated with vehicle, LPS, or LPS and CIAC001. Figure 6C shows Western blot results showing PKM2, H3, and GAPDH levels in the cytoplasm and nucleus of cells treated with vehicle, LPS, or LPS and CIAC001. Figure 6D shows relative LD levels in cells treated with vehicle, LPS, LPS and CIAC001, or LPS and compound 19b. Figure 6E shows HIF-1α levels in cells treated with vehicle, morphine, or morphine and CIAC001.

[0068] FIG7 shows the expression levels of M1 and M2 microglial mRNA markers in CIAC001- and LPS-treated cells.

[0069] Figures 8A-F show a set of schematic diagrams of animal behavioral experiments. Figure 8A is a schematic diagram for measuring changes in the number of jumps in mice induced by withdrawal syndrome after naloxone treatment and the therapeutic effect of CIAC001. Figure 8B is a schematic diagram for studying the effect of CIAC001 on morphine-induced behavioral sensitization. Figure 8C is a schematic diagram for describing the CPP test of mice after CIAC001 treatment. Figure 8D is a schematic diagram of the Y-maze, respiratory rate measurement, and open field test for evaluating the safety of CIAC001. Figure 8E is a schematic diagram of the Y-maze experiment for evaluating short-term memory impairment induced by CIAC001 or THC. Figure 8F is a schematic diagram of the CPP experiment for evaluating the rewarding effect of CIAC001.

[0070] Figures 9A-B show images of immunofluorescence staining of microglia. Figure 9A is a representative set of immunofluorescence images of NAc and VTA sections treated with vehicle, morphine, or morphine and CIAC001, showing Iba1-positive microglia. Cell nuclei were stained with DAPI. Figure 9B is a bar graph of the mean number of microglia per square millimeter in the NAc and VTA of mice treated with vehicle, morphine, or morphine and CIAC001.

[0071] FIG10 shows the pharmacokinetic profiles of CBD and CIAC001 in brain tissue (left) and plasma (right).

[0072] Figure 11A-C shows the RMSD Results and graphs of hydrogen bond counts from molecular dynamics simulations of PKM2, both with and without CIAC001, and structures generated from molecular dynamics simulations of PKM2 bound to CIAC001. Figure 11A is a graph of the RMSD of the PKM2 backbone generated from a 100 ns simulation, both with and without CIAC001. Figure 11B is a graph of the number of hydrogen bonds per frame between PKM2 dimer AB and dimer CD within the last 50 ns of a 100 ns molecular dynamics simulation, both with and without CIAC001. Figure 11C is a structure generated from a molecular dynamics simulation of PKM2 bound to CIAC001, showing hydrogen bonding interactions between dimer AB and dimer CD.

[0073] FIG12 shows a bar graph showing relative LDHA mRNA levels in BV-2 cells 3 h after treatment with vehicle, LPS, or LPS and CIAC001.

[0074] Figure 13 shows a graph of the concentration-effect curves of CIAC001 and CBD on forskolin-induced cAMP accumulation in HEK293 cells transfected with CB1 (left) or CB2 (right).

[0075] Figures 14A-F show graphs of memory and learning tests conducted on wild-type and Alzheimer's disease model mice. Escape latency in the Morris water maze test (14A), escape latency on the fifth day of water maze training (14B), exploration time in the platform quadrant area (14C) and number of platform crossings (14D) of mice on the first day after water maze training, and exploration time in the platform quadrant area (14E) and number of platform crossings (14F) of mice on the third day after water maze training.

[0076] Figures 14G-E show graphs of anxiety tests performed on wild-type and Alzheimer's disease model mice treated with vehicle or CIAC001. Number of grid crossings (14G), number of standing (14H), and number of defecation (14I) in the open field test. Novel object recognition index (14I) in the novel object recognition test. Total number of alternations (14K) and effective alternation rate (14L) in the Y-maze test. Number of open arm entries (14M) and time (14N) in the plus maze test.

[0077] 15A-F are Western blot results of Alzheimer's disease mouse models treated with different concentrations of CIAC001.

[0078] Figure 15A is a graph of potential measurements within the hippocampus. Figure 15B is a bar graph of potential measurements within the hippocampus shown in Figure 15A.

[0079] Figure 15C shows the Western blot results for Tau phosphorylation in the CIAC001-treated group. Figure 15D shows the Western blot results for TrkB phosphorylation, Syn1, and Aβ levels in the treated group. Figure 15E shows the grayscale values ​​and ratios of Tau5, p-Tau (T231), p-Tau (S202), and p-Tau (S404) proteins. Figure 15F shows the grayscale values ​​and ratios of p-TrkB (Y705), Syn1, Aβ1-42, and Aβ1-16 proteins. DETAILED DESCRIPTION

[0080] Before describing the present compounds, combinations, and methods, it is to be understood that this invention is not limited to the particular compounds, combinations, methods, and experimental conditions described, as such compounds, combinations, methods, and conditions may vary. Also, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of the present invention is limited only by the scope set forth in the appended claims.

[0081] All publications, patents, and patent applications mentioned herein are incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically indicated to be incorporated by reference.

[0082] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, it is understood that modifications and variations are encompassed within the spirit and scope of this disclosure. Preferred methods and materials are now described.

[0083] The present invention relates to the important discovery of CBD derivatives with therapeutic activity for neurodegenerative diseases and opioid addiction. These CBD derivatives are non-addictive, do not suppress breathing or impair short-term memory, have mild side effects and toxicity, and are safe for use in a variety of subjects and conditions.

[0084] In an embodiment, the present invention discloses a method for treating or preventing opioid addiction, withdrawal, or overdose, comprising administering a compound of formula I:

[0085] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures:

[0086] -H, or

[0087] R 2A and R 2Bare independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C4, wherein

[0088] R 2A and R 2B You can choose diazirine instead;

[0089] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0090] Subscript n ranges from 0 to 12;

[0091] thereby treating or preventing opioid addiction, withdrawal, or overdose.

[0092] In some cases, the compounds of formula I are narrowed down to compounds of formula Ia:

[0093] In some cases, R 1 Selected from the following structures: H, or In some cases, R 1 for

[0094] In some cases, R 2A and R 2B Each independently selected from the following structures: -H or C1-C4 alkane. In other cases, R 2A and R 2B are independently selected from the following structures: -H or -CH3. Or, R 2A and R 2B In some cases, n is 0 or 1; or n is 0.

[0095] In a special case, the compound of formula I is represented by

[0096] This invention includes methods for treating or preventing opioid addiction. Specifically, the invention includes methods for treating or preventing opioid withdrawal symptoms. Opioid withdrawal is one of the key symptoms of opioid addiction and is a key challenge in preventing drug cravings and relapse. While drugs such as proguanidine are commonly used to alleviate opioid withdrawal symptoms, these drugs also exhibit acute side effects and have limited effectiveness against residual opioid addiction. Compared to these drugs, the compounds disclosed herein have fewer side effects and exhibit significant efficacy in treating opioid addiction, thus providing an ideal alternative to existing treatments for opioid withdrawal. The compounds disclosed herein can be used to prevent or treat opioid withdrawal symptoms and are suitable for administration before, during, or after the onset of opioid withdrawal. The subjects referred to in this invention generally refer to subjects suffering from opioid withdrawal symptoms, or subjects suffering from acute opioid withdrawal symptoms. The compounds disclosed herein have low toxicity, and high doses can be used to combat severe withdrawal symptoms.

[0097] The compound of formula I can be administered over a wide range of dosages. For example, the compound of formula I can be administered at a dosage of about 0.5 μg / kg, 2 μg / kg, 20 μg / kg, 200 μg / kg, 2 mg / kg, 20 mg / kg, or 100 mg / kg. In the particular case of the present invention, the compound of formula I is administered at a dosage of about 0.5 mg / kg to 50 mg / kg, 0.5 to 10 mg / kg, 5 to 50 mg / kg, or 10 to 100 mg / kg. In other aspects, the compound of formula I can also be administered at a dosage of about 20 μg / kg to 500 μg / kg, 20 μg / kg to 200 μg / kg, 100 μg / kg to 500 μg / kg, or 250 μg / kg to 1 mg / kg. The compounds of Formula I can be administered multiple times per day (e.g., twice, three times, four times, five times, or six times per day), daily, weekly, monthly, or at any frequency in between (e.g., once every two, three, or four days). Because certain compounds of Formula I have brain and plasma half-lives of approximately 1 hour, in certain embodiments, the compounds of Formula I are administered one to ten times per day, one to six times per day, or two to four times per day.

[0098] In certain instances, the C of the compound of Formula I in the brain of the subject MAX In some cases, the brain to plasma ratio of the compound of Formula I is at least about 10:1, 25:1, 50:1, 100:1, or 200:1 at 1 hour after administration.

[0099] The compounds disclosed herein are suitable for a wide range of opioid addiction and withdrawal treatments. As non-limiting examples, opioids include fentanyl, ibuprofen, codeine, diazepam, dihydrocodeine, enkephalin, heroin, oxycodone, oxymorphone, pethidine, meprofen, methadone, morphine, nikomedin, opium, oscontin, oxycodone, oxymorphone, chenotetin, or derivatives, precursors, or pharmacologically acceptable salts or solvents thereof. In a specific aspect, the opioid is morphine. In a specific aspect, the opioid is heroin. In a specific case, the opioid is fentanyl. In the numerous aspects disclosed herein, the subject is addicted to an opioid. In a specific aspect, the subject exhibits symptoms of opioid addiction, including withdrawal reactions, behavioral sensitization, or conditioned place preference.

[0100] The compounds of formula I are suitable for a variety of routes of administration. As non-limiting examples, these compounds can be administered orally, subbuccally, sublingually, rectally, vaginally, intravenously, intraarterially, intramedullary, intramuscularly, intracerebrally, intracerebroventricularly, intraspinally, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, intramucosally, or by inhaler.

[0101] The compounds of formula I may be administered in admixture with pharmaceutical excipients, carriers and / or stabilizers. Examples of such ingredients are described in detail in Remington's Pharmaceutical Sciences, 18th edition (1990). Pharmaceutically acceptable carriers, excipients, or stabilizers may include buffers such as phosphate, acetic acid, and maleic acid; antioxidants such as ascorbic acid, ascorbyl palmitate, citric acid, methionine, tartaric acid, and vitamin E; preservatives such as octadecyldimethylbenzyl ammonium chloride, hexamethylammonium chloride, benzalkonium chloride, phenylalkonium chloride, and phenol; alkyl parabens such as p-methylbenzoate and p-hydroxybenzoate; resorcinol; low molecular weight polypeptides (e.g., peptides having approximately 10 or fewer amino acid residues); proteins such as serum albumin and collagen; amino acids such as glycine, glutamic acid, and lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; hydrophilic polymers such as polyvinylpyrrolidone and polyethylene glycol; sugars such as sucrose, mannitol, galactose, and trehalose; and / or nonionic surfactants such as glyceryl monostearate and sorbic monostearate. Examples of pharmaceutically acceptable carriers include, but are not limited to, ointments, particulate compositions, colloids, creams, gels, and emulsions. Examples of diluents include, but are not limited to, water, vegetable oils (such as soybean oil, sunflower oil, and palm oil), animal fats (such as fish oil and beef tallow), milk fat, and organic solvents (such as dimethyl sulfoxide).

[0102] In some embodiments, the invention further comprises administering an opioid antagonist to a subject. In certain embodiments, the opioid antagonist is naloxone or naltrexone. Without being limited by theory, the therapeutic efficacy of the compounds disclosed herein for opioid addiction and withdrawal may result from the synergistic effect of these compounds and opioid antagonists, alleviating opioid withdrawal symptoms and reducing the dosage requirement of the opioid antagonist.

[0103] While CBD exhibits anti-neuroinflammatory activity, its use in the context of this invention, including in the context of opioid therapy, is limited due to its neurotoxicity. Compared to CBD, the compounds disclosed herein offer a contrasting advantage to cannabinoids by exhibiting low toxicity and anti-neuroinflammatory activity. Thus, in many aspects, the compounds of Formula I do not induce toxic stress responses in subjects.

[0104] A sufficient dose of the compound of formula I can inhibit the overexpression of IL-1β, TNFα, IL-6 or a combination thereof in a subject induced by opioids (e.g., to an extent that cannot be achieved with CBD). The compound of formula I can regulate PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, or elongation factor 1γ in a subject when administered at a sufficient dose. In certain aspects, the compound of formula I can regulate PKM in a subject when administered at a sufficient dose. In certain special cases, PKM specifically refers to PKM2. The compound of formula I can increase the ratio of M2 to M1 microglia in the brain of a subject.

[0105] In the invention, the term "modulate" refers to an increase or decrease in the activity of a target. For example, in the case of a reference receptor protein, "modulate" can refer to an increase or decrease in one or more of signaling activity, cognate ligand binding affinity, ligand binding kinetics, or coreceptor binding. In the case of a reference enzyme, "modulate" can refer to an increase or decrease in enzyme activity. In some aspects, "modulate" refers to an increase in activity (e.g., agonist behavior produced by the compounds disclosed herein). In other aspects, "modulate" refers to a decrease in activity (e.g., antagonist behavior produced by the compounds disclosed herein).

[0106] In an embodiment, the present invention discloses a method for regulating PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, and elongation factor 1γ in a subject, comprising administering a compound of formula I:

[0107] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures: -H,

[0108] or

[0109] R 2A and R 2B are independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C4, wherein R 2A and R 2B You can choose to use diazirine instead;

[0110] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0111] Subscript n ranges from 0 to 12;

[0112] It thereby regulates PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, and elongation factor 1γ in the subject's body.

[0113] In a special case, the compound of formula I is

[0114] In particular cases, the methods involve regulating PKM2. In particular cases, the PKM is PKM2.

[0115] In certain aspects, the invention comprises modulating a PKM, in certain cases PKM2, in a subject.

[0116] The compounds disclosed in the present invention are very effective in regulating PKM2. PKM2 is a glycolytic rate-limiting enzyme and transcriptional coactivator distributed in tissues such as the brain and liver, and plays a key role in metabolic homeostasis and inflammation. PKM2 interacts with transcription factors (such as STAT3, Hif-1α, and ATF2) and promotes the production of pro-inflammatory cytokines such as IL-1β. Therefore, using the compounds disclosed in the present invention to target PKM2 is a new strategy for treating a range of diseases and symptoms, including neurodegenerative diseases, neuroinflammation, and opioid use disorder.

[0117] The present invention discloses a method for increasing the ratio of M2 to M1 microglia in the brain by administering a compound of formula I to a subject:

[0118] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures: -H,

[0119] or

[0120] R 2A and R 2B are independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C4, wherein R 2A and R 2B You can choose to use diazirine instead;

[0121] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0122] Subscript n ranges from 0 to 12;

[0123] This increased the ratio of M2 to M1 microglia in the subjects' brains.

[0124] Further, in the structure of the compound of formula I, n can be 0

[0125] In another aspect, the present invention provides a method for treating a neurodegenerative disease, comprising administering to a subject a compound of formula I:

[0126] or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures: -H,

[0127] or

[0128] R 2A and R 2Bare independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C4, wherein

[0129] R 2A and R 2B You can choose diazirine instead;

[0130] R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and

[0131] Subscript n ranges from 0 to 12;

[0132] thereby treating neurodegenerative diseases.

[0133] In certain aspects, the compound of formula I may preferably be a compound of formula Ia:

[0134] In special cases, R 1 Selected from the following structures: H, or More preferably, R 1 for

[0135] In some respects, R 2A and R 2B Each is independently selected from the following structures: -H or C1-C4 alkane. In other cases, R 2A and R 2B are independently selected from the following structures: -H or -CH3. Or, R 2A and R 2B All are -H.

[0136] In other cases, R 3A and R 3B Each of the following structures is independently selected: -H, -Cl, -Br, or -COOH. In some other cases, R 3A and R 3B All are -H. n is 0 or 1.

[0137] In special cases, the compound structure of formula I is

[0138] The compounds disclosed herein exhibit superior pharmacokinetic properties (particularly increased half-life and brain-to-plasma ratio), greater safety (including reduced cognitive impairment), lower toxicity, and similar anti-neuroinflammatory activity relative to CBD. Although CBD exhibits anti-neuroinflammatory activity, its toxicity has hindered its use in the treatment of neurodegenerative diseases. Surprisingly, it was discovered that shortening and functionalizing the side chains of CBD (CBD) mitigates the side effects (e.g., cognitive impairment) and toxicity of CBD without reducing its anti-neuroinflammatory activity, enabling the treatment of a wide range of previously untreatable neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis.

[0139] In many cases, the compounds of Formula I inhibit neuroinflammation in a subject. For example, the compounds of Formula I have an IC of 0.05 for inhibiting LPS-induced nitric oxide overproduction in microglia. 50 At most about 200 μM, 150 μM, 100 μM, 75 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, or at most about 1 μM.

[0140] The compounds of formula I may also have low toxicity. For example, the cytotoxicity IC of the compounds of formula I in microglia is 50 At least about 1 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 75 μM, 100 μM, 150 μM, or at least about 200 μM.

[0141] Formula I compound reduces the level of inflammatory markers in the subject. Specifically, the data disclosed in the present invention show that Formula I compound has a significant effect on reducing the expression of IL-1β, TNFα and IL-6 in microglia, and these three cytokines have a significant effect in promoting inflammation and promoting the pathological progression of neurodegenerative diseases. Therefore, in some aspects, the Formula I compound administered to the subject is sufficient to reduce the expression of IL-1β in microglia by at least about 1.5 times, 2 times, 2.5 times, 3 times, 4 times, 5 times, 6 times, 8 times, 10 times, 15 times, or at least about 20 times. At the same time, the Formula I compound administered to the subject is sufficient to reduce the expression of TNFα in microglia by at least about 1.5 times, 2 times, 2.5 times, 3 times, 4 times, 5 times, 6 times, 8 times, 10 times, 15 times, or at least about 20 times. In other aspects, the amount of the compound of Formula I administered to a subject is sufficient to reduce IL-6 expression in microglia by at least about 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold, or at least about 20-fold.

[0142] In specific aspects, the compound of Formula I at a dose of about 0.5 μg / kg, 2 μg / kg, 20 μg / kg, 200 μg / kg, 2 mg / kg, 20 mg / kg, or about 100 mg / kg can increase the C of the compound of Formula I in the brain of a subject. MAX One hour after administration to a subject, the brain-plasma drug concentration ratio of the compound of Formula I is at least about 10:1, 25:1, 50:1, 100:1, or at least about 200:1.

[0143] The compound of formula I is administered orally, subbuccal, sublingual, rectally, vaginally, intravenously, intraarterially, intraosseously, intramuscularly, intracerebrally, intraventricularly, submeningeal, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, submucosally, or by inhaler. Simultaneously, the compound of formula I can be administered together with an acceptable excipient.

[0144] Administration of a compound of Formula I is sufficient to modulate PKM, 60 kDa heat shock protein, α-enolase, eukaryotic translation elongation factor 1 α1, RNA helicase, heat shock protein 90-beta, ubiquitin carboxyl-terminal hydrolase, T-complex protein 1 subunit α, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor, clade B member 6a, or elongation factor 1-γ in a subject. In certain instances, the compound of Formula I is sufficient to modulate PKM in a subject. In certain instances, the PKM is specifically PKM2.

[0145] The administration of the compound of Formula I is sufficient to increase the ratio of M2 to M1 microglia in the brain of the subject. In another aspect, the compound of Formula I does not induce a toxic stress response in the subject.

[0146] The compound disclosed in the present invention is any one of compounds 1a-5a, 1b-5b, 12a-12h, CIAC001 or 19b-19f listed in Table 1.

[0147] Table 1

[0148] The following examples are intended to further illustrate embodiments of the present invention, but are not intended to limit the scope of the invention. Although they represent typical methods that may be used, other procedures, methods, or techniques familiar to those skilled in the art may also be used as alternatives.

[0149] Example 1

[0150] Experimental methods

[0151] Cell culture

[0152] BV-2 cells and GloSensor 22-F HEK293 cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C and 5% CO2.

[0153] Analytical reagents

[0154] 0.5 mg / mL 2,3-diaminonaphthalene: Dissolve 5 mg of 2,3,2-diaminonaphthalene in 10 mL of water containing 3 M HCl. Sonicate and vortex for 1 minute every 30 minutes until completely dissolved. Store the solution in a dark environment at -20°C.

[0155] 3M NaOH: 4g NaOH dissolved in 33mL H2O

[0156] 4% paraformaldehyde: Add 40g paraformaldehyde to 1L PBS (pH=7.4) and heat in 40℃ water for 1 day to dissolve

[0157] 0.05% crystal violet: Add 50 mg of crystal violet to 1 L of H2O and stir for 1 h.

[0158] 0.1mg / mL LPS: Dissolve 1mg LPS in 10mL ddH2O

[0159] 100 mM CIAC001: 3.69 mg CIAC001 dissolved in 100 μL DMSO

[0160] NO content analysis

[0161] BV-2 cells were plated at a density of 5 × 10 cells per well at 0.2 mL per well. 4 Cells were seeded in 96-well plates with 1 mL of culture medium (0.05 mg / mL) and incubated for 24 hours before being replaced with pure DMEM. 200 ng / mL LPS and various drug concentrations were added to each well. After 24 hours of treatment, 100 μL of culture medium was added to each well of the black 96-well plate. 10 μL of 0.05 mg / mL 2,3-diaminonaphthalene was added to each well. The reaction was incubated in the dark for 15 minutes, followed by termination with 5 μL of 3 M NaOH. NO content was measured by fluorescence at 430 nm under 360 nm excitation. Data were calculated by normalizing the fluorescence value to that of the LPS group.

[0162] Cytotoxicity assay

[0163] BV-2 cells were plated at a density of 5 × 10 cells per well at 0.2 mL per well. 4 / mL was planted in a 96-well plate and replaced with pure DMEM medium after culturing for 24 hours. Lipopolysaccharide (LPS, 200ng / mL) and different concentrations of drugs were added to each well, and the medium was removed after 24 hours of treatment. 100μL of 4% paraformaldehyde was added to each well and incubated for 5 minutes to fix the cells. After removing the paraformaldehyde, 100μL of 0.05% crystal violet was added and incubated at room temperature for 15 minutes. The cells were washed twice with clean water to remove the crystal violet, and finally 150μL of anhydrous ethanol was added and incubated at room temperature for 20 minutes. The absorbance at 540nm was measured using a multifunctional microplate reader, and the data were normalized to the blank group.

[0164] qRT-PCR

[0165] Total RNA was extracted from cells or mouse brain tissue using the RNeasy Mini Kit. The isolated RNA was reverse transcribed into cDNA using the RT2 Easy First Strand cDNA Synthesis Kit according to the manufacturer's protocol. PCR amplification was performed using synthetic primers, SYBR Green, and a TOptical Real-Time PCR Thermal Cycler (Analytik Jena, Thuringia, Germany). Data were analyzed using the ΔΔCt method, with Rpl7l1 or Rpl27 used as internal controls.

[0166] Cell morphology test

[0167] BV-2 cells were cultured in 1 mL per well at a cell density of 2.5 × 10 4 Cells were seeded in 6-well plates and cultured for 12 h before treatment with LPS (1 μg / mL) and 10 μM CIAC001 for 24 h. Cell morphology was observed and photographed using an inverted fluorescence microscope, and more than 300 cells were counted for each group.

[0168] Immunoblotting experiments

[0169] The protein samples were separated by SDS-PAGE, and the SDS-PAGE gel was used to transfer the separated proteins to the PVDF membrane under a constant voltage of 100V. Subsequently, the PVDF membrane was placed in a 5% skim milk powder solution and incubated at room temperature for 2 hours. The blocked PVDF membrane was added to the corresponding primary antibody skim milk powder solution and incubated at 4°C. After incubation for 12 hours, the PVDF membrane was washed 3 times with TBST solution, each time for 5 minutes. The washed PVDF membrane was incubated with the corresponding secondary antibody at room temperature for 1 hour, and then washed 3 times with TBST, each time for 5 minutes. The PVDF membrane was analyzed by a fully automatic chemiluminescence / fluorescence image analysis system and a chemiluminescent substrate (SuperSignal TM West Pico PLUS Chemiluminescent Substrate) was used for color development and imageJ software was used for processing.

[0170] cAMP content test

[0171] GloSensor 22-F HEK293 cells were transfected with CB1 and CB2 receptor plasmids, and 24 hours after transfection, the cells were cultured at 5×10 4 Cells were seeded at a density of 100 μg / mL in a 96-well plate. After 8 hours of culture, the culture medium was removed and DMEM supplemented with 2% GloSensor cAMP substrate was added for 2 hours. Finally, different concentrations of drugs and 10 mM forskolin were added to measure changes in chemiluminescence.

[0172] Cell target protein fishing experiment

[0173] BV-2 cells (confluence >90%) in 10 cm culture dishes were treated with CIAC001-P1 (5 μM) or CIAC001 (10 μM). After treatment at 37°C for 2 h, the dishes were transferred to an ice box and irradiated with 365 nm UV light for 30 min. The cells were washed twice with 2 mL of PBS, harvested using a cell scraper, and lysed in an ice-water bath for 30 min in 1 mL of cell lysis buffer (250 mM HEPES (pH 7.9), 150 mM KCl, 5 mM EDTA, 0.5% v / v NP-40, 1% v / v protease inhibitors). The cells were centrifuged at 12,000 g for 10 min at 4°C, and the supernatant was adjusted to a protein concentration of 1 mg / mL using a BCA assay kit. 1 mL of protein lysate was added to 80 μM biotin-N3, 100 μM TBTA, 1 mM TCEP, 1 mM CuSO4, and 5% v / v t-BuOH and allowed to react at room temperature for 1 hour. After the reaction, 20 μL of streptavidin magnetic beads (previously washed three times with 1 mL of PBS) were added and incubated at room temperature for 1.5 hours. The beads were then washed three times with 1 mL of PBS to remove nonspecific binding. 50 μL of 2× Laemmli buffer was then added and denatured at 100°C for 8 minutes. After separation on SDS-PAGE, the beads were stained with a silver staining kit, the gel was excised, and the target protein was identified by LC-MS.

[0174] In vitro target protein fishing assay

[0175] BV-2 cells were washed with PBS and then lysed with a cocktail containing 250 mM HEPES (pH 7.9), 1.5 M KCl, 5 mM EDTA, 0.5% v / v NP-40, and 0.025% v / v protease inhibitors and incubated on ice for 30 minutes. The lysate was diluted to a protein concentration of 1 mg / mL and incubated with the probe compound 19f (10 μM) for 2 hours. The lysate was subjected to the click reaction as described above and analyzed by SDS-PAGE. The SDS-PAGE gel was silver stained. The bands were excised and the proteins were identified by LC-MS / MS.

[0176] Cell thermal migration assay

[0177] According to 4×10 5 / mL density, plant BV-2 cells in a 10cm culture dish and incubate for 24h. After washing with PBS, collect the cells, add 1mL of cell lysis buffer (P0013, Biyuntian) containing 1% protease inhibitors and lyse in an ice water bath for 20min. Centrifuge at 12000g, 4℃ for 15min, take the supernatant, add 50μM CIAC001 and incubate at 20℃ for 2h. The blank group added the corresponding amount of DMSO as a control. After incubation, the protein lysate was divided into 60μL portions, incubated at different temperatures of 37-72℃ for 5min, and cooled at room temperature for 3min. Centrifuge at 20000g, 4℃ for 20min, take 40μL of supernatant, add 40μL of 2× Laemmli buffer, denature at 100℃ for 8min, and then perform protein immunoblotting experiment.

[0178] Fluorescence titration

[0179] PKM2 protein (0.5 μM) was titrated with increasing concentrations of CIAC001, and fluorescence intensity (λex 280 nm; λem 337 nm) was recorded using a fluorescence spectrophotometer (Cary Eclipse, Agilent Technologies Inc., USA). Appropriate controls were subtracted from the spectra obtained for the samples. The data were fitted by nonlinear least squares using Equation 8, derived from Equations 1-7. P T =[P]+2[LP2] (3) L T =[L]+[LP2] (4) F=F0-F PL ×[LP2] (5)

[0180] Because [LP2] 3 Much smaller than (L T +PT )x[LP2] 2 , Formula 6 can be simplified to Formula 7, and the calculation simplification is Formula 8:

[0181] Where F is the test fluorescence value; F0 is the initial PKM2 protein solution fluorescence value; F PL is the adjustment factor; K D is the binding constant; L T is the total concentration of CIAC001; P T is the total concentration of PKM2 protein. The stoichiometric ratio n is fitted by the following formula: lg(F0 / F-1)=-lg(K D )+n X lg([CIAC001]).

[0182] Molecular dynamics simulation

[0183] The structures of CIAC001 and compound 19b were drawn using GaussView 6 (GaussView) and optimized using Gaussian 09 (Gaussian 09) software under the B3LYP / 6-31G(d,p) basis set. The three-dimensional structure of the PKM2 tetramer was extracted from the crystal structure (PDB ID: 3ME3), and the activator in the structure was removed before docking. The missing hydrogen atoms were added using Maestro at pH 7.0. For molecular docking, The box completely covers the PKM2 tetramer. During docking, the protein was treated as rigid, while the ligand was treated as semiflexible. Based on their affinity for the PKM2 tetramer, the docking pose with the best binding affinity for PKM2 was selected from ten docking poses generated and ranked by AutoDock Vina.

[0184] The interaction between PKM2 and CIAC001 was analyzed by LigPlot + and PyMol (PyMol) software analysis.

[0185] The binding of CIAC001 to PKM2 tetramer was further studied by molecular dynamics simulation. The optimal docking position configuration was selected as the initial structure. The complex was constructed using a TIP3P water molecule solvent of 150 mM KCl on the CHARMM-GUI web server. The simulation was performed using the CHARMM36m force field and Gromacs 2021.2 program. The system was equilibrated for 100 ns in an isothermal-isobaric (NPT) set at 310.15 K. The bonds involving hydrogen atoms were constrained using the LINCS algorithm. The particle mesh Ewald (PME) summation method was used to treat long-range electrostatic interactions. The temperature (310.15 K) and pressure (1 atm) were maintained by the Nosé-Hoover Langevin piston and Parrinello-Rahman barostat methods. rmsd (root mean square deviation) analysis was performed by MDAnalysis.

[0186] PKM2 nuclear-cytoplasmic fractionation experiment

[0187] BV-2 cells were grown at a cell density of 5 × 10 4 The cells were inoculated with 100 μM LPS / mL in 10 cm culture dishes. After 24 h of culture at 37°C, the medium was replaced and treated with 200 ng / mL LPS and 10 μM CIAC001 for 6 h. The cells were washed with 3 mL of PBS to remove residual medium, harvested with a cell scraper, and centrifuged at 800 g for 5 min at 4°C. The supernatant was removed and 0.75 mL of cytoplasmic lysis buffer (10 mM HEPES pH 7.6, 10 mM KCl, 0.5 mM β-mercaptoethanol, 1.5 mM MgCl2, and 1 mM DTT, with 1% v / v protease inhibitors) was added and incubated in an ice-water bath for 20 min, vortexing every 5 min for 30 s. After lysis, 15 μL of 50% NP40 was added and the cells were lysed in an ice-water bath for 2 min. The cells were centrifuged at 12,000 g for 10 min at 4°C, and the supernatant was collected as cytoplasmic protein. The remaining pellet was resuspended and washed in 1 mL of PBS and centrifuged at 12,000 g for 10 minutes at 4°C to remove residual cytoplasmic proteins. This process was repeated twice to remove residual cytoplasmic proteins. The pellet was then resuspended in 100 μL of low-salt nuclear lysis buffer (10 mM Tris-HCl, pH 7.6, and 2 mM MgCl2) and lysed on ice for 20 minutes. The pellet was vortexed for 30 seconds every 5 minutes and centrifuged at 12,000 g for 15 minutes at 4°C to obtain the supernatant for nuclear proteins. The cytoplasmic and nuclear protein solutions were added to 2× Laemmli sample buffer, denatured at 100°C for 8 minutes, and then analyzed by western blotting.

[0188] ATP content test

[0189] BV-2 cells were collected at 5×104 Cells were seeded at a density of 1 mL / well in a 6-well plate and incubated at 37°C for 24 hours. After replacing the culture medium, 10 μM CIAC001 or 10 μM 19b was added and incubated for 2 hours. Intracellular ATP concentration was measured according to the instructions of the Enhanced ATP Kit (Beyotime, S0027-4).

[0190] Extracellular lactate content test

[0191] 5×10 4 BV-2 cells were seeded in 6-well plates at a density of 100 μg / mL and cultured at 37°C for 24 h. After replacing the culture medium, 200 ng / mL LPS, 10 μM CIAC001 or 10 μM 19b were added and incubated for 12 h. After incubation, 1 mL of culture medium was taken, centrifuged at 5000 g at 4°C for 5 min, and the supernatant was collected. The lactic acid concentration in the supernatant was tested according to the instructions of the lactic acid kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute).

[0192] Animal husbandry

[0193] Male Balb / c mice (20-28 g), aged 6-8 weeks, were purchased from Liaoning Changsheng Biotechnology Co., Ltd. Mice were housed in an animal room at 21-25°C, with lights on at 8:00 AM and off at 8:00 PM daily. Mice had free access to food and water throughout the period of housing. Experiments were performed after at least one week of acclimatization.

[0194] For behavioral experiments of Alzheimer's disease (Morris water maze, open field test, novel object recognition, Y-maze and elevated plus maze), the triple transgenic mouse model (B6:129-Psen1tm1Mpm Tg(APPswe, tauP301L)1Lfa / J, 3×Tg) was purchased from Jax Laboratories in the United States. Wild-type mice (C57BL / 6J, WT) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. 45 days after intraperitoneal injection of CIAC001, all mice underwent subsequent behavioral experiments. In order to minimize the use of mice, the mice were first tested for sufficient physical strength and normal mental state. The following schedule was followed: Day 1: open field test; Days 2-3: novel object recognition; Day 3: elevated plus maze; Day 4: Y-maze; Days 5-13: Morris water maze.

[0195] Triple transgenic model mice (B6:129-Psen1tm1Mpm Tg(APPswe, tauP301L)1Lfa / J, 3×Tg) were purchased from Jax Laboratories in the United States; wild-type mice (C57BL / 6J, WT) were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. and grouped according to Table 2.

[0196] Table 2

[0197] WT is a wild-type mouse, i.e., a wild-type mouse; 3×Tg-AD is a triple transgenic memory AD model mouse, i.e., the transgenic in the accompanying figure.

[0198] Morphine withdrawal experiment

[0199] Mice were injected with morphine three times daily for three consecutive days (5 mg / kg on day one, 20 mg / kg on day two, and 40 mg / kg on day three), with a 6-hour interval between injections. At 8:00 AM on day four, mice were injected with 40 mg / kg of morphine. After a 3-hour wait, they were injected with various concentrations of CIAC001. Thirty minutes later, naloxone (15 mg / kg) was injected. The number of jumps per 15 minutes was recorded.

[0200] Morphine behavioral sensitization experiment

[0201] Mice were placed in an open field (40×40×40 cm 3 ) for 3 days, 1 hour per day. After completing the adaptation, the mice were injected with morphine (10 mg / kg, ip), morphine (10 mg / kg, ip) + CIAC001 (0.2 mg / kg, ip), or morphine (10 mg / kg, ip) + CBD (0.2 mg / kg, ip) once a day for the first 7 consecutive days. The mice were then placed in the open field to explore freely for 1 hour. The mouse movement trajectory was recorded and the walking distance was analyzed. No drugs were given to the mice for the next 7 days. On the 15th day, the mice were injected with 5 mg / kg of morphine. The mouse movement trajectory was recorded within 1 hour and the movement distance was analyzed.

[0202] Morphine CPP experiment

[0203] Days 1-2: Mice were placed in the conditioned preference box for 15 minutes daily. Day 3: Mice were allowed to freely explore the conditioned preference box for 15 minutes. Their movements were recorded and the time spent in the black and white boxes was analyzed. The box in which the mice spent the most time was designated the preferred box, while the box in which they spent the most time was designated the non-preferred box. Days 4, 6, and 8: Mice were injected with either morphine (10 mg / kg, ip) or morphine (10 mg / kg, ip) plus CIAC001 (0.2 mg / kg, ip). Mice were placed in the non-preferred box for 30 minutes, the exit was sealed, and the mice were prohibited from freely moving around. Days 5, 7, and 9: Mice were injected with either saline (5 mL / kg, ip) or saline (5 mL / kg, ip) plus CIAC001 (0.2 mg / kg, ip). Mice were placed in the preferred box for 30 minutes, the exit was sealed, and the mice were prohibited from freely moving around. On day 10, mice were placed in the conditioned preference box and allowed to explore freely for 15 minutes. Their movements were recorded, and the time they spent in the black and white boxes was analyzed. The CPP score was calculated by subtracting the time spent in the preference box from the time spent in the non-preferred box.

[0204] Immunofluorescence experiments

[0205] After the CPP experiment was completed, the mice were sacrificed and perfused with normal saline. The brain tissue was removed, embedded in paraffin, and cut into 5 μm pieces using a vibrating microtome. The brain tissue sections were dewaxed in xylene and reactivated with ethanol solution. The brain tissue sections were placed in 0.01 M citrate buffer (pH 6.0) and heated in a microwave oven for antigen retrieval. Goat serum was added and incubated at 37°C for 20 minutes, followed by incubation with rabbit anti-Iba1 monoclonal antibody at 4°C overnight. After washing three times with PBS, the sections were incubated with Alexa-488 secondary antibody for 1 hour in the dark. Finally, the sections were washed twice with PBS for 5 minutes each time, counterstained with DAPI, and observed under a confocal microscope.

[0206] Y-maze

[0207] On days 1-2, mice were placed in a Y-maze (consisting of three interconnected, closed arms) and acclimated for 10 minutes. Afterwards, they were injected with CIAC001 (20 mg / kg) or THC (10 mg / kg). On day 3, mice were injected with CIAC001 or THC 45 minutes beforehand. Mice were then placed in one arm of the Y-maze. An object of different size and color was placed in either arm of the Y-maze. One arm was then sealed, prohibiting entry to the mice and marked as the "new object side." The remaining arm containing the object was marked as the "old object side." Mice were placed in the closed arm without the object and allowed to freely explore for 5 minutes before being transferred to a cage. After a 2-minute wait, the mice were placed in the Y-maze again. The new object side was opened and allowed to freely explore for 2 minutes. Movement patterns were recorded, and the time spent on the new and old object sides was analyzed. At the end of each experiment, the Y-maze was cleaned with 70% alcohol to prevent the contamination of the experiment with mouse odor.

[0208] Open field test

[0209] For the first two days of the experiment, mice were placed in an open field for 1 hour daily for acclimation. On the first day of the experiment, mice were allowed to freely explore the open field for 1 hour. Their movements were recorded, and their distance traveled, grooming, and scratching were analyzed. This served as the pre-test group. For the next 7 days, mice were injected with 20 mg / kg CIAC001 daily. 24 hours after the final injection, mice were allowed to freely explore the open field for 1 hour. Their movements were recorded, and their distance traveled, grooming, and scratching were analyzed. This served as the test group.

[0210] Respiratory rate test

[0211] Mice were injected daily with 20 mg / kg CIAC001 or 10 mg / kg THC. One hour after injection on days 2 and 5, mice were anesthetized with isoflurane and their respiratory rate within 20 seconds was measured. The average of three measurements per mouse was used.

[0212] Pharmacokinetic testing

[0213] Mice were administered CIAC001 (10 mg / kg, po) or CBD (10 mg / kg, po) dissolved in a 5% DMSO / 10% Solutol / 85% Captisol (20%) solution. Mice were sacrificed 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, and 4 hours after administration. Blood and brain tissue samples were collected and extracted with equal volumes of acetonitrile. The levels of CIAC001 and CBD in the blood and brain tissue were determined by HPLC-MS.

[0214] Chemical synthesis

[0215] Chemicals and solvents were purchased from Anaiji Chemical (Shanghai, China) unless otherwise stated. Analytical thin-layer chromatography (TLC) and silica gel were from Qingdao Siyuan Silicon Technology Co., Ltd. Flash chromatography was from Biotage. Silica gel columns were from Changzhou Santai Technology Co., Ltd. Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AV-300 spectrometer (300 MHz). 1 H, 75MHz 13 C), using CDCl3 or CD3OD solutions. In all cases, chemical shifts are expressed in ppm using the residual solvent as an internal reference (CDCl3: 1 H 7.26ppm, 13 C 77.16ppm; CD3OD: 1 H 3.31ppm, 13 C 49.03 ppm). Signal splitting patterns are described as chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, m = multit, br = broad), coupling constant and integration in Hertz (Hz). Low-resolution electrospray ionization (ES) mass spectra were recorded on a Waters QDa mass spectrometer, operating in positive and negative modes. The optical rotation of the CBD analogs was measured at 20°C using a polarimeter (Shanghai Jiahang Instrument Co., Ltd., Digipol-P610). The enantiomeric purity of the CBD analogs was measured using an OD-H chiral column (CHIRALCEL, ODHOCE-AP039). All compounds were >95% pure.

[0216] Water maze experiment

[0217] The Morris water maze is mainly composed of a circular pool with a diameter of about 160 cm and a depth of 50 cm, an underwater platform, and a data acquisition system. In the computer video analysis software, the water maze is artificially divided into four quadrants, and patterns of different colors and shapes are correspondingly posted on the four walls of the water maze pool. The platform in the water is a 25 cm 2 A circular piece of plexiglass was placed in the center of one quadrant of the pool, 15 cm from the edge and 1 cm below the water surface. A camera was installed directly above the pool to record and collect relevant data such as the mouse's movement trajectory, speed, and time. During the experiment, the pool was covered with curtains on all sides. The water in the pool was maintained at a depth of approximately 30 cm and a temperature of 22 ± 1°C. Milk powder was added to mask the mouse's odor and interfere with the mouse's vision. All experiments were conducted at the same time of day as much as possible.

[0218] Positioning navigation training: The experiment on days 1 to 5 is the positioning navigation test period. Pinch the mouse's tail and place it on the platform to adapt to the environment for 15 seconds. Guide the mouse to observe the pattern on the wall of a quadrant of the pool, and then gently place the mouse in the water of that quadrant. The maximum swimming time is set to 60 seconds. If the mouse finds the platform before 60 seconds, the mouse is immediately removed from the pool; if the mouse still does not find the platform after 60 seconds, the mouse is gently guided to the platform to make it remember again, and stay for 10 seconds. Then, it is taken out of the pool, placed next to a warm lamp to dry its hair and maintain body temperature, and placed in a mouse cage. The mice are tested in three quadrants every day, with an interval of 60 minutes between each experiment. The time it takes for the mouse to find the platform is recorded, which is the escape latency of the mouse.

[0219] Spatial Exploration Test: Spatial exploration tests were conducted 24 and 72 hours after the navigation. Short-term memory was tested at 24 hours, and long-term memory at 72 hours. The specific experimental method differed from the previous days in that the platform was removed from the pool and the mice were placed in the quadrant opposite the platform. The software recorded the number of times the mice crossed the original platform's location and the time they spent in the platform quadrant.

[0220] Recognition of new objects

[0221] The novel object recognition test involved mice freely exploring a 40cm x 40cm walled environment. The first day was a training period. Two identical objects (same color and shape) were placed symmetrically in the environment. The mice were then placed with their backs facing the objects and allowed to explore freely for 5 minutes. On the second day, one of the objects was replaced with a novel object (different color and shape). The mice were then placed with their backs facing the objects. The number of interactions (touching or approaching the objects) with the two objects was recorded over a 5-minute period. A camera positioned directly above the mice recorded the number and duration of these interactions.

[0222] Cross elevated maze

[0223] The elevated plus maze consists of two open arms (25×5×0.5 cm) that cross each other and are perpendicular to two closed arms (25×5×16 cm), with a platform (5×5×0.5 cm) in the center. The entire device is 50 cm above the floor.

[0224] At the beginning of the experiment, the mouse was placed in the center area of ​​the elevated plus maze, with its head facing one of the closed arms. The mouse was allowed to freely explore for 5 minutes throughout the experiment. A camera connected to a computer captured the mouse's movement data, recording the number and duration of entries into the open and closed arms. After each experiment with a mouse, all arms and the center area were cleaned with 75% alcohol and wiped dry with absorbent paper to prevent the odor of mouse secretions from interfering with subsequent experiments.

[0225] Detection of synaptic long-term potentiation in mouse brain

[0226] First, mice were stunned with isoflurane and sacrificed by cervical dislocation. The intact brain was then removed and quickly placed in pre-chilled artificial cerebrospinal fluid (ACSF) for 3 minutes, with oxygen continuously circulated to maintain brain activity. The brain was then quickly glued to the stage of a vibratome (olfactory bulb facing up, hindbrain facing down). The brain was then sliced ​​at 300 μm / slice and 0.14 mm / s in chilled ACSF purged with oxygen. The slices were collected starting at the point where the hippocampus was visible. The slices were then cut midway to separate the left and right hemispheres and incubated in a 32°C waterbath purged with oxygen for 2 hours. The computer recording software was then debugged to ensure a stable baseline. The incubated slice was placed between the electrodes, with the brightest line in the CA1 region of the hippocampus crossing the diagonal line of the baseline. The slices were then secured in place in a potential recorder using a grid and iron block. Perform small stimulation on each electrode potential, find the best stimulation point, record the stimulation point, then perform high-frequency stimulation at the stimulation point, and then use software to record the potential changes of each electrode point.

[0227] Tau pathology, Aβ pathology, and synapse-related protein indicators in mouse brain

[0228] Western blot technology was used to detect the protein contents of Tau5, p-Tau (T231), p-Tau (S202), p-Tau (S404), p-TrkB (Y705), Syn1, Aβ1-42, and Aβ1-16 in mouse hippocampal protein samples

[0229] Detection of insulin signaling pathway-related indicators related to AD pathology

[0230] Experimental process of mouse glucose tolerance (GTT) test: After fasting for 16 hours, the mice were weighed and the blood glucose level baseline was measured by tail incision using a blood glucose meter. Then, 2 g / kg glucose was injected intraperitoneally, and the glucose level was measured 30, 60, 90, 120 and 150 minutes after the glucose injection.

[0231] Insulin tolerance (ITT) experimental process: After fasting for 6 hours, the mice were weighed and the baseline blood glucose level was measured by tail incision using a glucometer. Then, 0.75 U / kg recombinant human insulin was injected intraperitoneally, and the glucose level was measured 30, 60, 90, 120 and 150 minutes after the injection of insulin.

[0232] Sample preparation

[0233] The compound (eg, CIAC001) was dissolved in 88% saline, 7% Tween 80, and 5% dimethyl sulfoxide. The prepared solution can be stored at 4°C for 2 weeks.

[0234] Example 2

[0235] Synthesis routes of compounds 1a-5a and 1b-5b

[0236] Methyl-4-pentyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2-ol (1a) and (1R,2R)-2′,6′-Dimethoxy-5-methyl-4′-pentyl-2-(prop-1-en-2-yl)-1,2,3,4-tetrahy-dro-1,1′-biphenyl (1b). 0.31 g (1 mmol) of CBD and 1.5 mmol (1.5 equiv) of CH3I were dissolved in 4 mL of acetone. 0.14 g (1 mmol, 1.0 equiv) of K2CO3 was ground into a powder and then added to the reaction solution. The solution was stirred at 60 degrees Celsius overnight. The reaction was quenched with 3 M HCl, and the aqueous phase was then extracted with ethyl acetate (2×20 mL). The organic phase was washed with brine and dried over Na2SO4. The crude product was purified by preparative thin layer chromatography to give a yellow oil. Product 1a: yellow oil, 0.085 g, 25.9%, 1 H NMR (300MHz, CDCl3): δ=6.32 (s, 1H), 6.24 (s, 1H), 6.03 (s, 1H), 5.59 (s, 1H), 4.50 (s, 1H), 4.33 (s, 1H), 4.01 (d, J=8.6Hz, 1H), 3.72 ( s, 3H), 2.57-2.37 (m, 3H), 2.28-2.09 (m, 2H), 1.80 (s, 5H), 1.68 (s, 3H), 1.63-1.56 (m, 2H), 1.38-1.30 (m, 4H), 0.90 (t, J=6.8Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=158.1, 155.7, 147.3, 142.7, 124.5, 115.0, 110.9, 109.5, 103.1, 55.5, 46.7, 36.0, 31.6, 30.9, 30.3, 28.1, 23.7, 22.6, 14.1. [a] D 20=-3 (c=1 mg / mL, CHCl3), ee(%)=99.0%. MS(ESI + )calcd for C 22 H 32 O2+Na + , 351.24 [M+Na]; found, 351.22. Product 1b: yellow oil, 0.075 g, 21.9%, 1 H NMR (300MHz, CDCl3): δ6.35 (s, 2H), 5.22 (s, 1H), 4.45 (d, J=4.7Hz, 2H), 4.00 (d, J=12.1Hz, 1H), 3.75 (s, 6H), 2.97-2.86 (m, 1H) , 2.59-2.51 (m, 2H), 2.29-1.92 (m, 2H), 1.81-1.73 (m, 3H), 1.68 (s, 3H), 1.62 (s, 4H), 1.39-1.30 (m, 4H), 0.92 (t, J=6.7Hz, 3H). 13 C NMR (75MHz, CDCl3): δ149.6, 141.9, 131.2, 125.9, 118.9, 109.6, 55.9, 45.2, 36.4, 36.1, 31.7, 31.1, 30.7, 29.7, 23.5, 22.6, 19.0, 14.1. [a]D 20 =-115 (c=1 mg / mL, CHCl3), ee(%)=99.7%. MS(ESI + )calcd for C 23 H 34 O2+H + , 343.22[M+H]; found, 343.27.

[0237] (1′R,2′R)-6-Ethoxy-5′-methyl-4-pentyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2-ol(2a) and (1R,2R)-2′,6′-Diethoxy-5-methyl-4′-pentyl-2-(prop-1-en-2-yl)-1,2,3,4-tetrahydro-1,1′-biphenyl(2b). 2a-2b were synthesized using CBD (0.31 g, 1 mmol) and CH3CH2Br (1.5 mmol, 1.5 equivalents) according to the synthetic procedures of 1a and 1b. Product 2a: yellow oil, 0.094 g, 27.5%, 1H NMR (300MHz, CDCl3): δ = 6.31 (s, 1H), 6.22 (s, 1H), 6.01 (s, 1H), 5.59 (s, 1H), 4.51 (s, 1H), 4.36 (s, 1H), 4.09-3.84(m, J=18.4Hz, 17.5, 9.4, 3H), 2.55-2.38(m, 3H), 2.30-1.98(m, 2H), 1.87-1.74(m, 4H), 1.69 (s, 3H), 1.66-1.54 (m, 3H), 1.41-1.28 (m, 7H), 0.90 (t, J=6.6Hz, 3H). 13 CNMR (75MHz, CDCl3): δ=157.4, 155.7, 147.2, 142.6, 124.7, 115.1, 111.0, 109.3, 103.8, 63.6, 46.4, 36.0, 31.6, 30.9, 30.3, 23.7, 22.6, 14.9, 14.1. [a] D 20 =-40 (c=1 mg / mL, CHCl3), ee(%)=98.8%. MS(ESI + )calcd for C 23 H 34 O2+Na + , 365.26 [M+Na]; found, 365.29. Product 2b: yellow oil, 0.071 g, 19.2%, 1 H NMR (300MHz, CDCl3): δ=6.30 (s, 2H), 5.25 (s, 1H), 4.45 (s, 1H), 4.40 (s, 1H), 4.06-3.84 (m, 5H), 2.93 (dd, J=16.3Hz, 9.1, 1H), 2. 56-2.46 (m, 2H), 2.28-1.93 (m, 3H), 1.82-1.73 (m, 2H), 1.70-1.64 (m, 5H), 1.58 (s, 3H), 1.40-1.30 (m, 9H), 0.90 (t, J=6.7Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=149.5, 141.7, 130.9, 126.3, 118.9, 109.7, 45.2, 36.4, 35.9, 31.7, 31.0, 30.7, 29.7, 29.5, 23.5, 22.6, 19.1, 15.0, 14.1. [a] D 20 =-29 (c=1 mg / mL, CHCl3), ee(%)=98.8%. MS(ESI+ )calcd for C 25 H 38 O2+Na + , 393.29[M+Na]; found, 393.31.

[0238] (1′R,2′R)-5′-Methyl-4-pentyl-2′-(prop-1-en-2-yl)-6-propoxy-1′,2′,3′,4′-tetrahydro-[1,1′-biphe nyl]-2-ol(3a) and (1R,2R)-5-Meth-yl-4′-pentyl-2-(prop-1-en-2-yl)-2′,6′-dipropoxy-1,2,3,4-tetrahydro-1,1′-biphenyl(3b). Compounds 3a and 3b were synthesized using CBD (0.31 g, 1 mmol) and CH3CH2CH2Br (1.5 mmol, 1.5 equivalents) according to the procedures 1a and 1b. The product 3a was obtained as yellow oil, 0.081 g, 22.7%, 1 H NMR (300MHz, CDCl3): δ=6.31(s, 1H), 6.23(s, 1H), 6.04(s, 1H), 5.60(s, 1H ), 4.53 (s, 1H), 4.39 (s, 1H), 4.05 (d, J = 7.8Hz, 1H), 3.92-3.76 (m, 2H), 2.56 -2.39(m, 3H), 2.33-2.01(m, 2H), 1.85-1.73(m, 7H), 1.68(s, 3H), 1.64-1. 55 (m, 2H), 1.41-1.29 (m, 4H), 1.03 (t, J=7.4Hz, 3H), 0.90 (t, J=6.7Hz, 3H). 13 C NMR (101MHz, CDCl3): δ=149.6, 141.7, 130.7, 126.8, 118.6, 109.6, 107.3, 105.0, 1 00.0, 44.9, 36.4, 36.2, 31.7, 31.1, 30.8, 29.7, 23.5, 22.9, 22.6, 19.3, 14.1, 10.8. [a] D 20 =-28 (c=1 mg / mL, CHCl3), ee(%)=99.9%. MS(ESI + )calcd for C 24 H 36 O2+Na +, 379.27 [M+Na]; found, 379.31. The product 3b was obtained as a yellow oil, 0.089 g, 22.1%, 1 H NMR (300MHz, CDCl3): δ=6.30 (s, 2H), 5.26 (s, 1H), 4.51-4.39 (m, 2H), 4.04 (d , J=12.5Hz, 1H), 3.84(dd, J=12.4Hz, 6.2, 4H), 3.06-2.87(m, 1H), 2.56-2.44( m, 2H), 2.33-1.91 (m, 3H), 1.83-1.72 (m, 5H), 1.66 (s, 3H), 1.64-1.60 (m, 3H), 1.58 (s, 2H), 1.38-1.31 (m, 4H), 1.04 (t, J=7.4Hz, 6H), 0.91 (t, J=6.8Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=149.6, 141.7, 130.7, 126.7, 118.5, 109.6, 44.9, 36.5, 36.2, 31.7, 31.1, 30.8, 29.6, 23.5, 22.9, 22.6, 19.3, 14.1, 10.8. [a] D 20 =-48 (c=1 mg / mL, CHCl3), ee(%)=99.9%. MS(ESI + )calcd for C 27 H 42 O2+Na + , 421.32[M+Na]; found, 421.34.

[0239] (1′R,2′R)-6-(Allyloxy)-5′-methyl-4-pentyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2-ol(4a) and (1R,2R)-2′,6′-Bis(allyloxy)-5-methyl-4′-pentyl-2-(prop-1-en-2-yl)-1,2,3,4-tetrahy-dro-1,1′-biphenyl(4b). Compounds 4a and 4b were synthesized using CBD (0.31 g, 1 mmol) and 3-bromoprop-1-ene (1.5 mmol, 1.5 equivalents) according to the procedures 1a and 1b. Product 4a was obtained as a yellow oil, 0.10 g, 28.2%, 1H NMR (300MHz, CDCl3): δ=6.33 (s, 1H), 6.23 (s, 1H), 6.04 (m, 2H), 5.60 (s, 1H), 5.39 (dd, J=17.3, 1.6Hz, 1H), 5.25 (dd, J=10.5, 1.4Hz, 1H), 4.54-4.33 ( m, 4H), 4.06 (d, J=8.4Hz, 1H), 2.57-2.39 (m, 3H), 2.34-2.01 (m, 3H), 1.85- 1.76 (m, 4H), 1.69-1.53 ​​(m, 5H), 1.38-1.29 (m, 4H), 0.90 (t, J=6.8Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=157.0, 155.8, 147.3, 142.6, 133.8, 124.6, 116.9, 111. 1, 109.7, 104.2, 100.0, 69.3, 36.0, 31.5, 30.8, 30.3, 29.7, 23.7, 22.6, 14.1. [a] D 20 =-22 (c=1 mg / mL, CHCl3), ee(%)=99.8%. MS(ESI + )calcd for C 24 H 34 O2+Na + , 377.26 [M+Na]; found, 377.27. The product 4b was obtained as a yellow oil, 0.070 g, 18.0%, 1 H NMR (300MHz, CDCl3): δ=6.35 (s, 2H), 6.12-5.96 (m, 2H), 5.45 (s, 1H), 5.40 ( s, 1H), 5.32 (s, 1H), 5.29-5.21 (m, 2H), 4.53-4.42 (m, 6H), 4.09 (d, J=8.7Hz , 1H), 3.05-2.91(m, 1H), 2.60-2.48(m, 2H), 2.28-1.95(m, 2H), 1.83-1.75( m, 2H), 1.68 (s, 3H), 1.66-1.55 (m, 5H), 1.33 (m, 4H), 0.92 (t, J=6.8Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=149.4, 141.8, 133.9, 131.0, 126.5, 119.4, 116.4, 10 9.9, 69.4, 45.1, 36.4, 36.2, 31.6, 31.0, 30.8, 29.6, 23.5, 22.6, 19.3, 14.1. [a]D 20 =-47 (c=1 mg / mL, CHCl3), ee(%)=99.9%. MS(ESI + )calcd for C 27 H 38 O2+Na + , 417.29[M+Na]; found, 417.29.

[0240] (1′R,2′R)-5′-Methyl-4-pentyl-2′-(prop-1-en-2-yl)-6-(prop-2-yn-1-yloxy)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2-ol(5a) and (1R,2R)-5-Methyl-4′-pentyl-2-(prop-1-en-2-yl)-2′,6′-bis(prop-2-yn-1-yloxy)-1,2,3,4-tetrahydro-1,1′-biphenyl(5b). Compounds 5a and 5b were synthesized using CBD (0.31 g, 1 mmol) and 3-Bromopropyne (1.5 mmol, 1.5 equivalents) according to the procedures of 1a and 1b. Product 5a was obtained as a yellow oil, 0.090 g, 25.6%, 1 H NMR (300MHz, CDCl3): δ=6.36 (s, 1H), 6.31 (s, 1H), 6.07 (s, 1H), 5.59 (s, 1H), 4.62-4.56 (m, 2H), 4.51 (s, 1H), 4.33 (s, 1H), 4.02 (d, J=8.4H z, 1H), 2.59-2.36 (m, 4H), 2.33-1.98 (m, 2H), 1.85-1.77 (m, 4H), 1.70 (s, 3H), 1.66-1.53 ​​(m, 3H), 1.40-1.28 (m, 4H), 0.90 (t, J=6.7Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=156.3, 155.8, 148.0, 142.7, 124.3, 115.7, 111.2, 110 .4, 104.4, 79.0, 74.9, 56.4, 46.6, 35.9, 31.5, 30.8, 30.3, 23.8, 22.6, 14.1. [a] D 20 =-23 (c=1 mg / mL, CHCl3), ee(%)=96.0%. MS(ESI + )calcd for C24 H 32 O2+Na + , 375.24[M+Na]; found, 375.22. Obtained product 5b: yellow oil, 0.095g, 24.3%, 1 H NMR (300MHz, CDCl3): δ=6.46 (s, 2H), 5.23 (s, 1H), 4.61 (d, J=2.3Hz, 4H), 4.48-4.42 (m, 2H), 4.01 (d, J=10.5Hz, 1H), 2.97-2.83 (m, 1H), 2. 61-2.51 (m, 2H), 2.48 (t, J=2.4Hz, 2H), 2.31-1.92 (m, 2H), 1.81-1.73 (m, 2H), 1.72-1.55 (m, 8H), 1.39-1.30 (m, 4H), 0.91 (t, J=6.8Hz, 3H). 13 C NMR (75MHz, CDCl3): δ=149.2, 141.9, 131.7, 125.3, 120.7, 110.1, 79.2, 74.6, 56.6, 45.4, 36.2, 36.1, 31.5, 30.9, 30.6, 29.7, 29.4, 23.5, 22.6, 19.1, 14.1. [a] D 20 =-71 (c=1 mg / mL, CHCl3), ee(%)=96.8%. MS(ESI + )calcd for C 27 H 34 O2+Na + , 413.26[M+Na]; found, 413.25.

[0241] Example 3

[0242] Synthesis route of compounds 12a-12h

[0243] This example covers the synthesis of CBD analogs with alkyl side chains of varying lengths. Briefly, olefin derivatives 8a-8e were prepared from triphenylphosphine bromide analogs 7a-7e and 3,5-dimethoxybenzaldehyde (6) via a Wittig reaction. Subsequently, olefin derivatives 8a-8e were hydrogenated and reduced with 10% Pd / C to afford 9b-9f. The methoxy group of 9b-9f was then removed by BBr3 to afford phenol analogs 11b-11f. Specifically, analog 11a was obtained by nucleophilic addition with CH3Li, followed by Pd / C-catalyzed hydrogenation and finally removal of the phenolic hydroxyl group. Menthadienol and olivetol derivatives 11a-11f were subjected to Friedel-Crafts alkylation to afford CBD derivatives 12a-12h of varying lengths.

[0244] Propyl-triphenylphosphonium Bromide (7a). Ph3P (2.62 g, 10 mmol) and bromopropane (1.13 g, 10 mmol, 1 equivalent) were dissolved in 50 mL of anhydrous toluene and refluxed for 18 h. After the reaction was complete, the reaction solution was cooled to room temperature and then 200 mL of ether was added and mixed evenly. The mixture was allowed to stand at room temperature for 30 min, and then filtered and dried to obtain the product 7a: 3.49 g, 63.0%, mp = 248-258 ° C. 1 H NMR (300MHz, CDCl3): δ=7.98-7.63 (m, 15H), 3.97-3.73 (m, 2H), 1.74 (s, 2H), 1.35-1.13 (m, 3H).

[0245] Pentyltriphenylphosphonium Bromide (7b). Compound 7b was synthesized by Ph3P (11.80 g, 40 mmol) and bromobutane (6.04 g, 40 mmol, 1 equivalent) according to the synthetic process of 7a. The product 7b was obtained as a white powder, 7.48 g, 60.5%, mp = 171-176, 1 H NMR (300MHz, CDCl3): δ=7.89-7.56 (m, 15H), 3.73-3.51 (m, 2H), 1.54 (dd, J=9.0, 5.4Hz, 4H), 1.23 (dt, J=14.0, 7.0Hz, 2H), 0.73 (t, J=7.3Hz, 3H).

[0246] Hexyltriphenylphosphonium Bromide (7c). Compound 7c was synthesized by Ph3P (11.80 g, 40 mmol) and bromopentane (6.60 g, 40 mmol, 1 equivalent) according to the process of 7a. The product 7c was obtained as a yellow oil, 7.65 g, 59.5%, 1 H NMR (300MHz, CDCl3): δ=7.79 (m, 15H), 3.89-3.77 (m, 2H), 1.70 (m, 4H), 1.25 (s, 4H), 0.83 (t, J=7.0Hz, 3H).

[0247] Heptyltriphenylphosphonium Bromide (7d). Compound 7d was synthesized by Ph3P (11.80 g, 40 mmol) and bromohexane (7.16 g, 40 mmol, 1 equivalent) according to the synthesis process of 7a. The product 7d was obtained as a yellow oil, 6.27 g, 41.0%, 1 H NMR (300MHz, CDCl3): δ=7.88-7.56 (m, 15H), 3.71-3.48 (m, 2H), 1.54 (d, J=3.6Hz, 4H), 1.27-1.00 (m, 6H), 0.72 (t, J=6.9Hz, 3H).

[0248] Hendecyltriphenylphosphonium Bromide (7e). Compound 7e was synthesized by Ph3P (11.80 g, 40 mmol) and bromododecane (9.40 g, 40 mmol, 1 equivalent) according to the synthesis process of 7a. The product 7e was obtained as a yellow oil, 8.75 g, 57.9%, 1 H NMR (300MHz, CDCl3): δ=7.73 (m, 15H), 3.67 (s, 2H), 1.58 (d, J=3.5Hz, 4H), 1.18 (d, J=16.0Hz, 14H), 0.81 (t, J=6.7Hz, 3H).

[0249] (Z) and (E)-1-(But-1-en-1-yl)-3,5-dimethoxybenzene (8a). The reaction mixture 7a (0.50 g, 1.3 mmol, 1.3 equivalents) was added to 20 mL of anhydrous tetrahydrofuran, and n-butyl lithium (1.3 mmol, 1.3 equivalents) was slowly added dropwise at -30°C. The mixture was stirred for 30 min until the reaction solution turned orange-red. 3,5-Dihydroxybenzaldehyde (0.14 g, 1 mmol) was then dissolved in 3 mL of anhydrous tetrahydrofuran and added dropwise to the reaction solution. After mixing well, the reaction solution was heated to 70°C and reacted for 16 h. After the reaction was completed, the reaction solution was cooled to -20°C and H2O was slowly added dropwise to quench the reaction. The aqueous phase was extracted with ether, and the organic phase was washed with saturated brine. The organic phase was dried over anhydrous Na2SO4 and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain product 8a, which was a mixture of E / Z. The product 8a was obtained: 0.15 g, 76.5%, E / Z ca. 70 / 30, (E)-8a: 1 H NMR (300MHz, CDCl3): δ=6.50 (d, J=2.3Hz, 2H), 6.36-6.34 (m, 1H), 6.37-6.32 (m, 2H), 3.78 (s, 6H), 2.17-2.26 (m, 2H), 1.08 (t, J=8.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=161.0, 140.2, 133.4, 129.0, 104.2, 99.3, 55.5, 26.2, 13.8. (Z)-8a: 1 H NMR (300MHz, CDCl3): δ6.43=(d, J=2.2Hz, 2H), 6.3-6.24 (m, 2H), 5.65 (m, 1H), 3.80 (s, 6H), 2.34 (m, 2H), 1.08 (t, J=8.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.7, 139.8, 135.4, 128.4, 107.0, 106.6, 98.9, 55.5, 22.3, 14.6. MS(ESI + )calcd for C 12 H 16 O2+H + , 193.12[M+H] + ;found,193.04.

[0250] (Z) and (E)-1-(Hex-1-en-1-yl)-3,5-dimethoxybenzene (8b). Compound 8b was synthesized from 7b (1.71 g, 2.6 mmol, 1.3 equiv) and compound 6 (0.33 g, 2 mmol) according to the synthetic process of 8a. The product was an E / Z mixture. Product 8b was obtained: 0.33 g, 74.5%, E / Z ca. 65 / 35, (E)-8b: 1 H NMR (300MHz, CDCl3): δ = 6.49 (d, J=2.2Hz, 2H), 6.36-6.30 (m, 1H), 6.37-6.12 (m, 2H), 3.78 (s, 6H), 2.15-2.22 (m, 2H), 1.52-1.24 (m, 4H), 0.91 (t, J=7.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=161.1, 140.2, 132.0, 129.9, 104.2, 99.3, 55.5, 32.8, 31.7, 22.5, 14.2. (Z)-8b: 1 H NMR (300MHz, CDCl3): δ6.42=(d, J=2.2Hz, 2H), 6.36-6.30 (m, 2H), 5.64 (m, 1H), 3.78 (s, 6H), 2.32 (m, 2H), 1.52-1.24 (m, 4H), 0.91 (t, J=7.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.7, 139.9, 133.9, 129.9, 128.9, 107.0, 98.9, 55.5, 32.3, 28.7, 22.6, 14.2. MS(ESI + )calcd for C 14 H 20 O2+H + ,221.15[M+H] + ;found,221.07.

[0251] 1-(3,5-Dimethoxyphenyl)-1-heptylene (8c). Compound 8c was synthesized from 7c (1.11 g, 2.6 mmol, 1.3 equiv) and compound 6 (0.33 g, 2 mmol) according to the synthesis process of 8a. The product was an E / Z mixture. Product 8c was obtained: 0.35 g, 75.0%, E / Z ca. 63 / 37, 1 H NMR, (E)-8c: 1H NMR (300MHz, CDCl3): δ6.53 (d, J=2.1Hz, 2H), 6.38-6.35 (m, 1H), 6.31-6.19 (m, 2H), 3.8 2(s, 6H), 2.27-2.17(m, 2H), 1.54-1.43(m, 2H), 1.38-1.30(m, 4H) 0.93(t, J=7.2Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=161.1, 140.2, 132.1, 129.8, 104.2, 99.3, 55.3, 33.1, 31.6, 29.2, 22.8, 14.3. (Z)-8c: 1 H NMR (300MHz, CDCl3): δ=6.46 (d, J=2.2Hz, 2H), 6.38-6.30 (m, 2H), 5.73-5.64 (m , 1H), 3.82 (s, 6H), 2.39-2.30 (m, 2H), 1.38-1.30 (m, 4H), 0.93 (t, J=7.2Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.7, 138.4, 131.7, 128.8, 126.9, 107.1, 95.8, 55.5, 33.0, 31.8, 29.4, 22.8, 14.3. MS(ESI + )calcd for C 15 H 22 O2+H + ,235.16[M+H] + ;found,235.09.

[0252] 1-(3,5-Dimethoxyphenyl)-1-octylene (8d). Compound 8d was synthesized from compound 7d (1.15 g, 2.6 mmol) and compound 6 (0.33 g, 2 mmol) according to the synthetic process of 8a. The product was an E / Z mixture. Product 8d: 0.35 g, 71.0%, E / Z ca. 56 / 44. 1 H NMR, (E)-8d: 1 H NMR (300MHz, CDCl3): δ=6.53 (d, J=2.1Hz, 2H), 6.39-6.35 (m, 1H), 6.32-6.19 (m, 2H), 3.8 2(s, 6H), 2.25-2.18(m, 2H), 1.52-1.42(m, 2H), 1.40-1.25(m, 6H) 0.90(t, J=7.0Hz, 3H); 13C NMR (75MHz, CDCl3): δ=161.1, 140.2, 132.1, 129.8, 104.2, 99.3, 55.5, 33.2, 30.1, 29.3, 29.0, 22.8, 14.3. (Z)-8d: 1 H NMR (300MHz, CDCl3): δ=6.46 (d, J=2.1Hz, 2H), 6.39-6.31 (m, 2H), 5.73-5.64 (m, 1H), 3.8 2(s, 6H), 2.39-2.32(m, 2H), 1.52-1.42(m, 2H), 1.40-1.25(m, 6H) 0.90(t, J=7.0Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.7, 140.2, 134.4, 128.9, 107.0, 98.9, 55.5, 33.2, 33.0, 29.5, 29.1, 22.8, 14.3. MS(ESI + )calcd for C 16 H 24 O2+H + ,249.08[M+H] + ;found,248.99.

[0253] 1-(3,5-Dimethoxyphenyl)-1-laurylene (8e) was synthesized from compound 7e (1.29 g, 2.6 mmol) and compound 6 (0.33 g, 2 mmol) according to the synthetic scheme of 8a. The product 8e was obtained: 0.35 g, 70.8% yield, E / Z ca. 64 / 36. 1 H NMR, (E)-8e: 1 H NMR (300MHz, CDCl3): δ=6.54 (d, J=2.1Hz, 2H), 6.39-6.35 (m, 1H), 6.32-6.19 (m, 2H), 3.82 (s, 6H), 2.25-2.18 (m, 2H), 1.52-1.42 (m, 2H), 1.40-1.25 (m, 14H) 0.90 (t, J=7.0Hz, 3H); (E)-8e: 13 C NMR (75MHz, CDCl3): δ160.9, 140.0, 131.9, 129.6, 104.0, 99.1, 55.3, 33.0, 31.9, 30.0, 29.6, 29.6, 29.4, 29.3, 28.8, 22.7, 14.1.). (Z)-8e: 1H NMR (300MHz, CDCl3): δ=6.45 (d, J=2.1Hz, 2H), 6.39-6.31 (m, 2H), 5.73-5.64 (m, 1H), 3.82 (s, 14H), 2.39-2.32(m, 2H), 1.52-1.42(m, 2H), 1.40-1.25(m, 6H) 0.91(t, J=7.0Hz, 3H); (Z)-8e: 13 C NMR (75MHz, CDCl3): δ=160.5, 139.7, 133.8, 128.7, 106.9, 98.7, 55.3, 33.0, 31.9, 30.0, 29.6, 29.6, 29.4, 29.3, 28.8, 22.7, 14.1. MS(ESI + )calcd for C 20 H 32 O2+H + , 305.24[M+H] + ;found,305.10.

[0254] 1-(3,5-Dimethoxyphenyl)Ethane (9a). Compound 10 (1 mmol) and 3 mL of acetic acid were dissolved in 20 mL of methanol, and then 20 mg of palladium on carbon was added. H2 was bubbled into the reaction solution at room temperature using a balloon until the reactant 10 was completely reacted. After the reaction was completed, the reaction solution was filtered to remove the palladium on carbon and concentrated to obtain product 9a: 0.15 g, 92.3% 1 H NMR (300MHz, CDCl3): δ=6.43 (d, J=2.3Hz, 2H), 6.36 (t, J=2.2Hz, 1H), 3.83 (s, 6H), 2.65 (q, J=7.6Hz, 2H), 1.29 (t, J=7.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.8, 146.8, 105.9, 97.6, 55.2, 29.3, 15.5. MS(ESI + )calcd for C 10 H 14 O2+H + , 195.13[M+H] + ;found,195.05.

[0255] 1-(3,5-Dimethoxyphenyl)Butane(9b). Compound 8a (1.0 mmol, 0.19 g) was dissolved in 10 mL of methanol, palladium on carbon was added, and H2 was bubbled into the reaction solution using a balloon at room temperature until the raw materials reacted completely. The reaction solution was then filtered to remove the palladium on carbon and concentrated to obtain product 9b: 0.18 g, 91.8%, 1 H NMR (300MHz, CDCl3): δ=6.34 (d, J=2.1Hz, 2H), 6.29 (t, J=2.2Hz, 1H), 3.77 (s, 6H), 2.62-2.4 5 (m, 2H), 1.58 (dt, J=15.4, 7.5Hz, 2H), 1.34 (dq, J=14.5, 7.3Hz, 2H), 0.91 (t, J=7.3Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.9, 145.6, 106.7, 97.7, 55.4, 36.2, 33.6, 22.6, 14.2. MS(ESI + )calcd for C 12 H 18 O2+H + , 195.13[M+H] + ;found,195.05.

[0256] 1-(3,5-Dimethoxyphenyl)Hexane (9c). Compound 9c was synthesized by 8b according to the synthesis process of 9b to obtain product 9c: 0.21g, 93.5%, 1 H NMR (300MHz, CDCl3): δ=6.38 (d, J=2.2Hz, 2H), 6.33 (t, J=2.2Hz, 1H), 3.81 (s, 6H), 2.62-2.52(m, 2H), 1.70-1.55(m, 2H), 1.35(t, J=8.7Hz, 6H), 0.92(t, J=6.7Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.9, 145.6, 106.6, 97.7, 55.4, 36.5, 31.9, 31.5, 29.2, 22.8, 14.3. MS(ESI + )calcd for C 14 H 22 O2+H + ,223.16[M+H] + ;found,223.07.

[0257] 1-(3,5-Dimethoxyphenyl)Butane (9d). Compound 9d was synthesized via 8c according to the synthesis process of 9b to obtain product 9d: 0.23 g, 97.1%, 1 H NMR (300MHz, CDCl3): δ=6.37 (d, J=2.2Hz, 2H), 6.32 (t, J =2.2Hz, 1H), 3.80 (s, 6H), 2.60-2.53 (m, 2H), 1.68-1.56 (m, 2H), 1.38-1.26 (m, 8H), 0.90 (t, J = 6.8Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.9, 145.6, 106.7, 97.7, 55.4, 36.5, 32.0, 31.5, 29.5, 29.4, 22.9, 14.3. MS(ESI + )calcd for C 15 H 24 O2+H + ,237.18[M+H] + ;found,237.09.

[0258] 1-(3,5-Dimethoxyphenyl)Octane (9e). Compound 9e was synthesized from 8d according to the synthesis process of 9b to obtain product 9e: 0.24g, 94.1%, 1 H NMR (300MHz, CDCl3): δ=6.34 (d, J=2.2Hz, 2H), 6.29 (t, J=2.2Hz, 1H), 3.77 (s, 6 H), 2.58-2.49 (m, 2H), 1.67-1.52 (m, 2H), 1.28 (m, 10H), 0.88 (t, J=6.7Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.7, 145.4, 106.5, 97.5, 55.2, 36.3, 31.9, 31.3, 29.5, 29.4, 29.3, 22.7, 14.1. MS(ESI + )calcd for C 16 H 26 O2+H + ,251.19[M+H] + ;found, 251.10.

[0259] 1-(3,5-Dimethoxyphenyl)Dodecane (9f). Compound 9f was synthesized via 8e according to the synthesis process of 9b to obtain product 9f: 0.29 g, 95.4%,1 H NMR (300MHz, CDCl3): δ=6.34 (d, J=2.2Hz, 2H), 6.29 (t, J=2.2Hz, 1H), 3.77 ( s, 6H), 2.59-2.49 (m, 2H), 1.58 (m, 2H), 1.27 (m, 18H), 0.87 (t, J=5.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.9, 145.6, 106.6, 97.7, 55.4, 36.5, 32.1, 31.5, 29.9, 29.9, 29.8, 29.7, 29.6, 22.9, 14.3. MS(ESI + )calcd for C 20 H 34 O2+H + , 307.26[M+H] + ;found,307.17.

[0260] 1-(3,5-Dimethoxyphenyl)Ethan-1-ol(10). Compound 6 (1.66 g, 10 mmol) was dissolved in 30 mL of anhydrous tetrahydrofuran, and then methyl lithium (12 mmol, 1.2 equivalents) was slowly added dropwise at -20°C. After reacting for 30 min, saturated NH4Cl was added dropwise at low temperature to quench the reaction. The aqueous phase was extracted twice with 50 mL of ethyl acetate, dried over Na2SO4, and concentrated to obtain product 10: 1.73 g, 95.12%, 1 H NMR (300MHz, CDCl3): δ=6.52 (d, J=2.2Hz, 2H), 6.36 (t, J=2.3Hz, 1H), 4.80 (q, J=6.4Hz, 1H), 3.79 (s, 6H), 2.53 (s, 1H), 1.47 (d, J=6.5Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.9, 148.3, 103.3, 99.2, 70.5, 55.4, 25.1. MS(ESI + )calcd for C 10 H 14 O3+Na + ,205.09[M+Na] + ;found,204.97.

[0261] 1-(3,5-Dihydroxyphenyl)Ethane (11a). Compound 9a (2 mmol) was dissolved in 50 mL of anhydrous dichloromethane, and 6 mL of BBr3 dichloromethane solution (1 mmol / mL, 3 equivalents) was slowly added dropwise at -20°C. After reacting at room temperature for 12 h, H2O was slowly added dropwise at -20°C to quench the reaction. The aqueous phase was extracted twice with 30 mL of ethyl acetate, and the organic phase was then washed with saturated NaHCO3, dried over anhydrous Na2SO4, and concentrated to obtain product 11a-: 0.24 g, 86.0%, 1 H NMR (300MHz, CDCl3): δ=6.28 (d, J=5.0Hz, 2H), 6.20 (t, J=2.1Hz, 1H), 4.82 (s, 2H), 2.55 (q, J=7.6Hz, 2H), 1.21 (t, J=7.6Hz, 3H); 13 C NMR (75MHz, CDCl3): 8=156.6, 147.5, 107.5, 100.1, 28.7, 15.2. MS(ESI + )calcd for C8H 10 O2+H + ,139.07[M+H] + ; found, 138.99.

[0262] 1-(3,5-Dihydroxyphenyl)Butane(11b). 11b Compound 11b was synthesized by 9b according to the synthesis process of 11a to obtain product 11b: 0.28g, 84.5%, 1 H NMR (300MHz, CDCl3): δ=6.24 (d, J=1.9Hz, 2H), 6.16 (t, J=2.0Hz, 1H), 2.38 (t, J=7 .7Hz, 2H), 1.52-1.37 (m, 2H), 1.25 (dq, J=14.3, 7.2Hz, 2H), 0.84 (t, J=7.3Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.2, 146.6, 108.6, 100.5, 35.7, 33.3, 22.5, 14.1. MS(ESI - )calcd for C 10 H 14 O2-H + , 165.10[MH] - ;found,165.01.

[0263] 1-(3,5-Dihydroxyphenyl)Hexane (11c). Compound 11c was synthesized from 9c according to the synthesis process of 11a to obtain product 11c: 0.37g, 94.1%, 1 H NMR (300MHz, CDCl3): δ=6.28 (d, J=2.1Hz, 2H), 6.20 (t, J=2.2Hz, 1H), 5.44 (s, 2H), 2.5 7-2.40 (m, 2H), 1.55 (dd, J=14.7, 6.9Hz, 2H), 1.34-1.25 (m, 6H), 0.89 (t, J=6.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.4, 146.6, 108.4, 100.5, 36.0, 31.9, 31.2, 29.2, 22.8, 14.3. MS(ESI - )calcd for C 12 H 18 O2-H + , 193.13[MH] - ;found,193.05.

[0264] 1-(3,5-Dihydroxyphenyl)Heptane (11d). Compound 11d was synthesized from 9d according to the synthesis process of 11a to obtain product 11d: 0.37 g, 89.6%, 1 H NMR (300MHz, CDCl3): δ=6.26 (d, J=2.2Hz, 2H), 6.20 (t, J=2.2Hz, 1H), 4.87 (s, 2H), 2.5 5-2.44 (m, 2H), 1.67-1.51 (m, 2H), 1.29 (dt, J=10.1, 4.4Hz, 8H), 0.89 (t, J=6.7Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.2, 146.4, 108.3, 100.3, 35.8, 31.8, 31.1, 29.3, 29.2, 22.7, 14.1. MS(ESI - )calcd for C 13 H 20 O2-H + , 207.15[MH] - ;found,207.06.

[0265] 1-(3,5-Dihydroxyphenyl)Octane (11e). Compound 11e was synthesized from 9e according to the synthesis process of 11a to obtain product 11e: 0.40 g, 90.1%, 1 H NMR (300MHz, CDCl3): δ=6.25 (d, J=1.9Hz, 2H), 6.16 (t, J=1.9Hz, 1H), 5.95 (s, 2 H), 2.39 (m, 2H), 1.68-1.48 (m, 2H), 1.34-1.23 (m, 10H), 0.86 (t, J=6.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.3, 146.7, 108.52, 100.5, 36.0, 32.1, 31.2, 29.7, 29.6, 29.5, 22.9, 14.3. MS(ESI - )calcd for C 14 H 22 O2-H + , 221.16[MH] - ;found,221.08.

[0266] 1-(3,5-Dihydroxyphenyl)Dodecane(11f). Compound 11f was synthesized from 9f according to the synthesis process of 11a to obtain product 11f: 0.49g, 87.7%, 1 H NMR (300MHz, CDCl3): δ=6.23 (d, J=2.0Hz, 2H), 6.17-6.12 (m, 1H), 5.02 (s, 2H ), 2.50-2.37(m, 2H), 1.58-1.44(m, 2H), 1.23(s, 18H), 0.86(t, J=6.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.5, 146.5, 108.4, 100.4, 36.1, 32.1, 31.3, 29.9, 29.9, 29.8, 29.8, 29.6, 22.9, 14.3. MS(ESI - )calcd for C 18 H 30 O2-H + , 277.22[MH] - ;found,277.14.

[0267] (1′R,2′R)-4-Ethyl-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-dio l(12a). Menthadienol (6 mmol, 2 equivalents), compound 11a (3 mmol) and formic acid (0.28 g, 6 mmol, 2 equivalents) were dissolved in 20 mL of dichloromethane and reacted at room temperature for 12 h. After that, saturated NaHCO 3 was added to quench the reaction. The aqueous phase was extracted twice with 20 mL of ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous Na 2 SO 4 and concentrated to obtain a crude product. The crude product was eluted with petroleum ether: ethyl acetate = 25:1. The product 12a: 0.49 g, 17.7%, was obtained by silica gel column chromatography. 1 H NMR (300MHz, CDCl3): δ = 6.33 (m, 2H), 6.11 (s, 1H), 5.57 (s, 1H), 5.30 (s, 1H), 4.66 (s, 1H), 4.57 (s, 1H), 3.94 ( d, J=6.5Hz, 2H), 2.58-2.38 (m, 3H), 2.37-2.04 (m, 2H), 1.89-1.78 (m, 5H), 1.71 (s, 3H), 1.18 (t, J=7.6Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.0, 144.3, 140.1, 124.2, 113.9, 111.0, 46.3, 36.8, 30.4, 28.4, 28.4, 23.8, 20.1, 15.1. MS(ESI - )calcd for C 18 H 24 O2-H + , 271.2[MH] - ;found,271.2,[a] D 20 =-55 (c=1.0 mg / mL, CHCl3), ee%=99.9%.

[0268] (1′R,2′R)-5′-Methyl-2′-(prop-1-en-2-yl)-4-propyl-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-diol (CBDV, 12b). Compound 12b was synthesized according to the synthetic process of compound 12a to obtain product 12b: 1H NMR (300MHz, CDCl3): δ=6.24 (m, 2H), 6.02 (s, 1H), 5.59 (s, 1H), 4.74 (s, 1H), 4.64 (d, J=8.6Hz, 2H), 3.87 (d, J= 7.0Hz, 1H), 2.51-2.34(m, 3H), 2.33-2.03(m, 2H), 1.89-1.74(m, 5H), 1.72-1.52(m, 5H), 0.92(t, J=7.3Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.4, 142.8, 140.1, 124.1, 113.8, 110.9, 46.2, 37.6, 37.2, 30.4, 28.4, 24.0, 23.7, 20.5, 13.8. MS(ESI - )calcd for C 19 H 26 O2-H + , 285.19[MH] - ;found,285.09. [a]D 20 =-59 (c=1.0 mg / mL, CHCl3), ee%=99.9%.

[0269] (1′R,2′R)-4-Butyl-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-dio l(12c). Compound 12c was synthesized according to the synthetic process of compound 12a to obtain product 12c: 0.22 g, 14.8%, colorless oil. 1 H NMR (300MHz, CDCl3): δ = 6.22 (m, 2H), 5.99 (s, 1H), 5.58 (s, 1H), 4.86 (s, 1H), 4.65 (s, 1H), 4.56 (s, 1H), 3.89 (d, J =8.5Hz, 1H), 2.53-2.33(m, 3H), 2.35-2.05(m, 2H), 1.91-1.74(m, 5H), 1.39-1.16(m, 7H), 0.93(t, J=7.3Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.3, 143.0, 140.1, 124.1, 113.7, 110.9, 46.2, 37.1, 35.2, 33.1, 30.4, 28.4, 23.7, 22.3, 20.4, 14.0. MS(ESI - )calcd for C20 H 28 O2-H + ,299.21;found[MH] - :299.16. [a] D 20 =-52 (c=1.0 mg / mL, CHCl3), ee%=98.5%.

[0270] (1′R,2′R)-5′-Methyl-4-pentyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-diol (CBD, 12d). Compound 12d was synthesized according to the synthetic process of compound 12a to obtain product 12d: 1 H NMR (300MHz, CDCl3): δ = 6.24 (m, 2H), 6.02 (s, 1H), 5.59 (s, 1H), 4.77 (s, 1H), 4.68 (s, 1H), 4.58 (s, 1H), 3.88(d, J=10.2Hz, 1H), 2.54-2.35(m, 3H), 2.35-2.03(m, 2H), 1.92-1.75(m , 5H), 1.68 (s, 3H), 1.64-1.50 (m, 2H), 1.41-1.23 (m, 4H), 0.91 (t, J=6.8Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.4, 143.1, 140.1, 124.1, 113.8, 110.9, 46.2, 37.2, 35.5, 31.5, 30.7, 30.4, 28.4, 23.7, 22.6, 20.5, 14.1.). MS(ESI - )calcd for C 21 H 30 O2-H + , 313.22[MH] - ; found, 313.17. [a] D 20 experimental=-130(c=10.0mg / mL, EtOH), (lit.[a] D 20 =-124 (10.0 mg / mL, EtOH), ee% = 99.9%.

[0271] (1′R,2′R)-4-Hexyl-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-diol (12e). Compound 12e was synthesized according to the synthetic process of compound 12a to obtain product 12e: 0.29 g, 17.6%, colorless oil. 1 H NMR (300MHz, CDCl3): δ = 6.22 (m, 2H), 5.99 (s, 1H), 5.58 (s, 1H), 5.18 (s, 1H), 4.65 (s, 1H), 4.56 (s, 1H), 3.90 (d, J = 11.3Hz, 1H), 2. 55-2.36 (m, 3H), 2.33-2.04 (m, 2H), 1.89-1.75 (m, 5H), 1.67 (s, 3H), 1.63-1.49 (m, 2H), 1.34-1.21 (m, 6H), 0.93 (t, J=7.0Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.2, 142.9, 140.0, 124.2, 113.8, 110.9, 58.5, 46.2, 37.0, 35.2, 33.1, 30.4, 28.4, 23.7, 22.4, 20.3, 18.4, 14.0. MS(ESI - )calcd for C 22 H 32 O2-H + , 327.24[MH] - ; found, 327.19. [a] D 20 =-61 (c=1.0 mg / mL, CHCl3), ee%=96.2%.

[0272] (1′R,2′R)-4-Heptyl-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-diol(12f). Compound 12f was synthesized according to the synthetic process of compound 12a to obtain product 12f: 0.25 g, 18.6%, colorless oil. 1H NMR (300MHz, CDCl3): δ=6.23 (m, 2H), 6.02 (s, 1H), 5.59 (s, 1H), 4.90 (s, 1H), 4.67 (s, 1H), 4.57 (s, 1H), 3.95-3.83 (m, 1H), 2 .52-2.35(m, 3H), 2.31-2.02(m, 2H), 1.90-1.74(m, 5H), 1.68(s, 3H), 1.63-1.50(m, 2H), 1.26(m, 8H), 0.91(t, J=7.2Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.3, 143.0, 140.0, 124.2, 113.7, 110.9, 46.2, 37.1, 35.5, 31.8, 31.0, 30.4, 29.3, 29.2, 28.4, 23.7, 22.7, 20.4, 14.1. MS(ESI - )calcd for C 23 H 34 O2-H + , 341.26[MH] - ; found, 341.27. [a] D 20 =-66 (c=1.0mg / mL, CHCl3), ee%=96.2%.

[0273] (1′R,2′R)-5′-Methyl-4-octyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetra-hydro-[1,1′-biphenyl]-2,6-diol (12g). Compound 12g was synthesized according to the synthetic process of compound 12a to obtain product 12g: 0.25g, 14.2%, as a colorless oil. 1 H NMR (300MHz, CDCl3): δ=6.26 (m, 2H), 6.04 (s, 1H), 5.59 (s, 1H), 4.95 (s, 1H), 4.65 (s, 1H), 4.55 (s, 1H), 3.88 (d, J=11.7Hz, 1H), 2.53-2.36(m, 3H), 2.34-2.03(m, 2H), 1.94-1.77(m, 5H), 1.68(s, 3H), 1.62-1.48(m, 2H), 1.28(s, 10H), 0.90(t, J=6.7Hz, 3H); 13C NMR (75MHz, CDCl3): δ=149.2, 143.0, 140.1, 124.1, 113.8, 110.9, 46.2, 37.1, 35.6, 31.9, 31.0, 30.4, 29.5, 29.4, 29.3, 28.4, 23.7, 22.7, 20.4, 14.2. MS(ESI - )calcd for C 24 H 36 O2-H + , 355.27[MH] - ; found, 355.24. [a] D 20 =-60 (c=1.0mg / mL, CHCl3), ee%=99.8%.

[0274] (1′R,2′R)-4-Dodecyl-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tet-rahydro-[1,1′-biphenyl]-2,6-diol(12h). Compound 12h was synthesized according to the synthetic process of compound 12a to obtain product 12h: 0.41 g, 18.6%, colorless oil. 1 H NMR (300MHz, CDCl3): δ6.28 (m, 2H), 6.02 (s, 1H), 5.59 (s, 1H), 4.75 (s, 1H), 4.68 (s, 1H), 4.58 (s, 1H), 3.89 (d, J=11.7Hz, 1H), 2.5 3-2.36 (m, 3H), 2.34-2.04 (m, 2H), 1.91-1.74 (m, 5H), 1.68 (s, 3H), 1.64-1.49 (m, 2H), 1.40-1.16 (m, 18H), 0.90 (t, J=6.7Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=149.4, 143.1, 140.1, 124.1, 113.7, 110.9, 46.2, 37.2, 35.5, 32.0, 31.0, 30.4, 29.7, 29.7, 29.6, 29.6, 29.4, 29.3, 28.4, 23.7, 22.7, 20.5, 14.2. MS(ESI - )calcd for C 28 H 44 O2-H + , 411.3[MH] - ; found, 411.4. [a] D20 =-74 (c=1.0 mg / mL, CHCl3), ee%=99.9%.

[0275] Example 4

[0276] Synthesis process of compounds 19b-19d

[0277] This example covers the synthesis of analogs with different side chain functional groups. Briefly, compound 14 was synthesized by the reaction of 3,5-dihydroxyacetophenone (13) and benzyl bromide, and intermediate 14 was then reduced to compound 15 by NaBH4. In the presence of phosphorus tribromide, the α-OH group of 15 was replaced by Br to generate intermediate 16. Nucleophilic substitution reactions with pyrrole, imidazole, and triazole produced 17a-17d, followed by benzyl deprotection to obtain 18a-18d. CBD derivatives 19a-19d were obtained by Friedel-Crafts alkylation of 18a-18d with menthadiol.

[0278] 1-(3,5-Bis(benzyloxy)phenyl)Ethan-1-one (14). Compound 13 (10 mmol, 1.52 g), benzyl bromide (25 mmol, 4.28 g, 2.5 eq.), and K2CO3 (10 mmol, 1.38 g, 1 eq.) were added to 60 mL of acetone. After reflux for 24 h, 5 mL of triethylamine was added and refluxed for 30 min. After completion of the reaction, 3 volumes of H2O were added, and the aqueous phase was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous Na2SO4, and concentrated to yield product 14: 3.25 g, 98%. 1 H NMR (300MHz, CDCl3): δ=7.53-7.33 (m, 10H), 7.24 (d, J=2.3Hz, 2H), 6.85 (t, J=2.3Hz, 1H), 5.10 (s, 4H), 2.59 (s, 3H); 13 C NMR (75MHz, CDCl3): δ=197.8, 160.0, 139.1, 136.4, 128.7, 128.2, 127.7, 107.4, 106.9, 70.4, 26.8. MS(ESI + )calcd for C 22 H 20 O3+H + , 333.14[M+H] + ;found,332.94.

[0279] 1-(3,5-Bis(benzyloxy)phenyl)Ethan-1-ol(15). Compound 14 (9.5 mmol, 3.17 g) was dissolved in 80 mL of DCM:MeOH = 20:1 (v:v) and NaBH4 (12 mmol, 0.45 g, 1.25 eq.) was slowly added at -20°C. The reaction was allowed to proceed for 30 min at -20°C and then for 1 h at room temperature. H2O was added to quench the reaction and the mixture was extracted three times with 70 mL of ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous Na2SO4, and concentrated to afford product 15: 3.14 g, 98.9%. 1 H NMR (300MHz, CDCl3): δ=7.51-7.31 (m, 10H), 6.66 (d, J=2.1Hz, 2H), 6.56 (t, J=2.2Hz, 1H), 5.05 (s, 4H), 4.83 (q, J=6.4Hz, 1H), 1.48 (d, J=6.4Hz, 3H); 13 CNMR (75MHz, CDCl3): δ=160.1, 148.6, 136.9, 128.7, 128.1, 127.7, 104.5, 101.0, 70.5, 70.1, 25.2. MS(ESI - )calcd for C 22 H 22 O3-H + , 333.16[MH] - ;found,332.94.

[0280] (((5-(1-Bromoethyl)-1,3-phenylene)bis(oxy))bis(methylene))-dibenzene(16). Compound 15 (9 mmol, 3.01 g) was dissolved in 200 mL of ether, and PBr3 (13.5 mmol, 1.5 eq.) was slowly added dropwise. The mixture was then refluxed for 5 h. After completion, saturated NaHCO3 was added dropwise at -20°C to quench the reaction. The aqueous phase was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous Na2SO4, and concentrated to yield product 16: 3.50 g, 98.2%. 1 H NMR (300MHz, CDCl3): δ=7.53-7.32 (m, 10H), 6.75 (d, J=2.1Hz, 2H), 6.59 (t, J=2.1Hz, 1H), 5.16 (q, J=6.9Hz, 1H), 5.07 (s, 4H), 2.05 (d, J=6.9Hz, 3H); 13C NMR (75MHz, CDCl3): δ=160.0, 145.5, 136.7, 128.7, 128.2, 127.7, 106.2, 101.8, 70.2, 49.5, 26.8. MS(ESI + )calcd for C 22 H 21 BrO2+H + ,397.07[M+H] + ;found,396.93.

[0281] (1-(3,5-Bis(benzyloxy)phenyl)ethyl)-2H-1,2,3-triazole(17a) and (1-(3,5-Bis(benzyloxy)phenyl)ethyl)-1H-1,2,4-triazole(17b). 2H-1,2,3-Triazole (12 mmol, 0.83 g, 1.5 eq.) was dissolved in anhydrous DMF, and NaH (15 mmol, 0.36 g, 1.87 eq.) was slowly added to the solvent at -20°C. After stirring for 30 min, compound 16 (8 mmol, 3.17 g) was dissolved in DMF and added to the reaction solution, followed by reaction at 110°C for 24 h. After completion of the reaction, saturated NH4Cl was added to terminate the reaction, and the organic phase was extracted with ether and washed with saturated brine, dried over anhydrous Na2SO4, and concentrated to obtain the crude product. The crude product was separated by silica gel column chromatography to obtain compound 17a: 0.80 g, 26.1%, 1 H NMR (300MHz, CDCl3): δ=7.62 (s, 2H), 7.46-7.28 (m, 10H), 6.51 (s, 3H), 5.79 (q, J=7.1Hz, 1H), 4.97 (s, 4H), 1.96 (d, J=7.1Hz, 3H); 13 CNMR (75MHz, CDCl3): δ=160.2, 143.3, 136.7, 134.2, 128.7, 128.2, 127.7, 105.7, 101.4, 70.2, 64.3, 21.3. MS(ESI + )calcd for C 24 H 23 N3O2+H + ,408.18[M+Na] + ; found, 408.26. Obtained 17b: 0.88 g, 28.5%. 1H NMR (300MHz, CDCl3): δ=7.66 (s, 1H), 7.45 (s, 1H), 7.41-7.25 (m, 9H), 6.58 (t, J=2.0Hz , 1H), 6.50 (d, J=2.0Hz, 2H), 5.73 (q, J=7.0Hz, 1H), 4.96 (s, 4H), 1.90 (d, J=7.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.0, 142.2, 136.3, 133.6, 128.4, 127.9, 127.4, 122.2, 105.6, 101.3, 69.8, 59.7, 21.0. MS(ESI + )calcd for C 24 H 23 N3O2+Na + ,408.18[M+Na] + ;found,408.27.

[0282] 1-(1-(3,5-Bis(benzyloxy)phenyl)ethyl)-1H-imidazole (17c). Compound 17c was synthesized from 16 (3.57 g, 9 mmol) and 1H-imidazole (0.92 g, 13.5 mmol) according to the synthetic process of 17a. 17c was obtained: 0.58 g, 16.7%, 1 H NMR (300MHz, CDCl3): δ=7.59 (s, 1H), 7.45-7.29 (m, 10H), 7.09 (s, 1H), 6.91 (s, 1H), 6.55 (t, J =2.2Hz, 1H), 6.37 (d, J = 2.1Hz, 2H), 5.25 (q, J = 6.9Hz, 1H), 4.98 (s, 4H), 1.81 (d, J = 7.0Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.4, 144.1, 136.5, 136.2, 129.5, 128.7, 128.2, 127.7, 118.1, 105.5, 101.2, 70.2, 56.6, 22.0. MS(ESI + )calcd for C 25 H 24 N2O2+H + , 385.19[M+H] + ;found,385.06.

[0283] 1-(1-(3,5-Bis(benzyloxy)phenyl)ethyl)-1H-pyrrole (17d). Compound 17d was synthesized from 16 (3.57 g, 9 mmol) and 1H-pyrrole (0.91 g, 13.5 mmol) according to the synthesis process of 17a. 17d was obtained: 0.69 g, 20.1%, 1 H NMR (300MHz, CDCl3): δ=7.59-7.39 (m, 10H), 6.88 (t, J=2.0Hz, 2H), 6.65 (t, J=2.0Hz, 1H), 6.49 (d , J=2.1Hz, 2H), 6.34 (t, J=1.9Hz, 2H), 5.29 (q, J=7.0Hz, 1H), 5.07 (s, 4H), 1.90 (d, J=7.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=160.1, 146.1, 136.7, 128.6, 128.1, 127.7, 119.5, 108.2, 105.3, 100.6, 70.0, 58.1, 22.0. MS(ESI + )calcd for C 26 H 25 NO2+Na + ,406.19[M+Na] + ;found,406.04.

[0284] 5-(1-(2H-1,2,3-Triazol-2-yl)ethyl)benzene-1,3-diol (18a). Compound 17a (1.2 mmol, 0.46 g) was dissolved in 20 mL of methanol, 20 mg of palladium on carbon was added, and H2 was bubbled into the reaction solution using a balloon until the reaction was complete. The palladium on carbon was filtered, dried over anhydrous Na2SO4, and concentrated to obtain product 18a: 0.20 g, 82.0%, 1 H NMR (300MHz, CDCl3): δ=7.58 (s, 2H), 7.12 (s, 2H), 6.15 (d, J=1.9Hz, 2H), 6.08 (t, J=2.0Hz, 1H), 5.68 (q, J=7.0Hz, 1H), 1.87 (d, J=7.1Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=157.2, 143.3, 134.2, 105.9, 102.8, 64.3, 21.0. MS(ESI - )calcd for C 10 H 11 N3O2-H +, 204.09[MH] - ;found,203.99.

[0285] 5-(1-(1H-1,2,4-Triazol-1-yl)ethyl)benzene-1,3-diol (18b). Compound 18b was synthesized according to the synthetic process of 18a to obtain 18b: 0.17 g, 85.6% as a colorless oil. 1 H NMR (300MHz, MeOD): δ=9.37 (s, 1H), 9.20 (s, 1H), 7.79 (t, J=2.1Hz, 1H), 7.75 (d, J=2.1Hz, 2H), 7.22 (q, J=7.0Hz, 1H), 6.52 (s, 2H), 3.37 (d, J=7.1Hz, 3H); 13 C NMR (75MHz, MeOD): δ=160.0, 144.2, 134.3, 124.8, 105.9, 103.4, 61.5, 21.6. MS(ESI - )calcd for C 10 H 11 N3O2-H + , 204.09[MH] - ;found,203.97.

[0286] 5-(1-(1H-Imidazole-1-yl)ethyl)benzene-1,3-diol (18c). Compound 18c was synthesized according to the synthetic process of 18a to obtain 18c: 0.21 g, 81.0% as a colorless oil. 1 H NMR (300MHz, MeOD): δ=7.74 (s, 1H), 7.08 (s, 1H), 6.93 (s, 1H), 6.11 (t, J=2.1 Hz, 1H), 6.06 (d, J=2.1Hz, 2H), 5.26 (q, J=7.0Hz, 1H), 1.71 (d, J=7.1Hz, 3H). MS(ESI - )calcd for C 11 H 12 N2O2-H + , 203.09[MH] - ;found,203.00.

[0287] 5-(1-(1H-Pyrrol-1-yl)ethyl)benzene-1,3-diol (18d). Compound 18d was synthesized according to the synthetic process of 18a to obtain 18d: 0.36 g, 79.6% as a colorless oil. 1 H NMR (300MHz, MeOD): δ=6.71 (t, J=2.1Hz, 2H), 6.18-5.98 (m, 5H), 5.10 (q, J=7.0Hz, 1H), 1.68 (d, J=7.1Hz, 3H); 13 C NMR (75MHz, MeOD): δ=159.7, 147.9, 120.5, 108.7, 105.7, 102.5, 59.2, 22.5. MS(ESI + )calcd for C 12 H 13 NO2+K + , 242.19[M+K] + ;found,242.18.

[0288] (1′R,2′R)-4-(1-(2H-1,2,3-Triazol-2-yl)ethyl)-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahy dro-[1,1′-biphenyl]-2,6-diol (19a, CIAC001). Compound 18a (0.5 mmol) was dissolved in 20 mL of dichloroethane, followed by the addition of p-toluenesulfonic acid (0.1 mmol, 0.02 g, 0.2 eq.) and (1S,4R)-1-methyl-4-(1-methylvinyl)-2-cyclohexen-1-ol (1 mmol, 0.15 g, 2 eq.). After reacting at room temperature for 2 h, the reaction was terminated by the addition of saturated NaHCO₃. The reaction was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated to obtain the crude product. The crude product was separated using silica gel column chromatography to obtain compound 19a: 0.04 g, 21.2%. 1 H NMR (300MHz, CDCl3): δ=7.49 (s, 2H), 6.22 (s, 1H), 5.96 (s, 2H), 5.71 (s, 1H), 5.65-5.55 (m, 1H), 5.40 (s, 1H), 4.47 (s, 1H), 4 .37(s, 1H), 3.79(d, J=9.6Hz, 1H), 2.35-2.22(m, 1H), 2.14-1.90(m, 2H), 1.85-1.75(m, 3H), 1.72-1.60(m, 5H), 1.54(s, 3H);13 C NMR (75MHz, CDCl3): δ=148.6, 140.6, 140.3, 134.0, 123.9, 116.5, 111.2, 77.4, 63.9, 46.2, 36.7, 30.4, 28.4, 23.8, 20.0. MS(ESI - )calcd for C 20 H 25 N3O2-H + , 338.19[MH] - ; found, 338.14. [a] D 20 =-56 (c=1.0 mg / mL, CHCl3), ee%=99.1%.

[0289] (1′R,2′R)-4-(1-(1H-1,2,4-Triazol-1-yl)ethyl)-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahyd ro-[1,1′-biphenyl]-2,6-diol (19b). Compound 19b was synthesized according to the synthetic process of 19a at 70°C to obtain 19b: 0.25 g, 16.9%. 1 H NMR (300MHz, CDCl3): δ=8.91 (s, 1H), 7.60 (s, 1H), 7.44 (d, J=2.3Hz, 1H), 6.43 -6.33(m, 2H), 6.18(s, 1H), 5.67(dq, J=14.0, 7.0Hz, 1H), 5.54(s, 1H), 4.54(s, 1H), 4.45 (s, 1H), 4.05 (d, J=8.8Hz, 1H), 2.45 (td, J=9.8, 5.4Hz, 1H), 2.18 (dt, J=37.9, 17.8Hz, 2H), 1.92 (t, J=7.5Hz, 3H), 1.82-1.76 (m, 5H), 1.68 (d, J=13.1 Hz, 3H); 13 C NMR (75MHz, CDCl3): δ=156.6, 148.0, 140.0, 139.0, 133.4, 124.2, 122.8, 11 7.1, 111.2, 106.2, 77.4, 60.4, 46.5, 30.5, 28.3, 23.8, 20.7, 19.4.) MS (ESI - )calcd for C 20 H 25 N3O2-H + , 338.19[MH]- ; found, 338.14. [α] D 20 =-68 (c=1.0 mg / mL, CHCl3), ee%=99.0%.

[0290] (1′R,2′R)-4-(1-(1H-Imidazole-1-yl)ethyl)-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2,6-diol (19c). Compound 19c was synthesized according to the synthetic process of 19a to obtain 19c: 0.27 g, 19.9%. δ=8.64(s,1H),8.32(s,1H),7.09(s,1H),6.37(s,2H),5.50(s,1H),5.35(m,1H),4.50(s,1H),4.42( s, 1H), 4.03 (d, J=7.0, 1H), 2.45 (td, J=10.0, 4.8, 1H), 2.21 (m, 2H), 1.91-1.74 (m, 8H), 1.68 (s, 3H). 13 C NMR (75MHz, CDCl3): δ=156.8, 147.9, 139.9, 137.4, 135.9, 123.7, 111.1, 77.2, 50.7, 30.3, 28.2, 23.7, 20.9, 19.1. MS(ESI - )calcd for C 21 H 26 N2O2-H + , 337.20[MH] - ; found, 337.09. [α] D 20 =-72 (c=1.0 mg / mL, CHCl3), ee%=99.7%.

[0291] (1′R,2′R)-4-(1-(1H-Pyrrol-1-yl)ethyl)-5′-methyl-2′-(prop-1-en-2-yl)-1′,2′,3′,4′-tetrahydro-[1,1′-biphenyl]-2,6-diol (19d). Compound 19d was synthesized according to the synthetic process of 19a to obtain 19d: 0.30 g, 27.9%. 1H NMR (300MHz, CDCl3): δ=6.75 (t, J=2.1Hz, 2H), 6.28 (s, 1H), 6.19 (t, J=2.1Hz, 2H), 6.01 (d, J=20.6Hz, 1H), 5.55 (s, 1H), 5.13 (q, J=7.0Hz, 1H), 4. 83 (s, 1H), 4.65 (dd, J = 5.9, 4.2Hz, 1H), 4.54 (s, 1H), 3.89 (d, J = 8.8Hz, 1 H), 2.41 (td, J=10.6, 3.8Hz, 1H), 2.18 (m, 2H), 1.79 (m, 8H), 1.67 (s, 3H); 13 C NMR (75MHz, CDCl3): δ=148.8, 148.8, 143.5, 143.4, 140.4, 123.7, 119.6, 1 15.6, 111.1, 107.9, 57.6, 46.1, 36.9, 30.4, 28.3, 23.7, 21.9, 20.2, 20.1. MS(ESI - )calcd for C 22 H 27 NO2H + , 336.20[MH] - ; found, 336.15. [α] D 20 =-77 (c=1.0 mg / mL, CHCl3), ee%=99.9%.

[0292] Example 5

[0293] Structure-activity relationship of compounds 1a-5a and 1b-5b

[0294] This example covers the structure-activity relationship of CBD derivatives with different functional group-substituted hydroxyl groups. The phenolic hydroxyl group of CBD plays a key role in its biological activity and cytotoxicity. This includes the preparation of CBD phenolic derivatives by synthesizing compounds 1a-5a and 1b-5b via different alkyl halides, as described in Example 2.

[0295] To quantitatively investigate the in vitro anti-neuroinflammatory activity of CBD derivatives, the BV-2 microglial cell line was used because it closely resembles primary microglia in morphology, phenotype, and function. Microglia are immune cells of the central nervous system. Activation of microglia leads to the release of proinflammatory cytokines, promoting neuroinflammation.

[0296] The anti-neuroinflammatory activity of monoalkylated CBD derivatives (compounds 1a-5a) was demonstrated by inhibiting the overproduction of pro-inflammatory cytokine nitric oxide induced by LPS in BV-2 cells with a half-maximal inhibitory concentration (IC 50) to reflect (see Table 3). With the replacement of the phenolic hydroxyl group with a single alkyl group, the anti-neuroinflammatory activity of these compounds was significantly reduced. CBD derivatives 1b-5b with two phenolic hydroxyl groups alkylated showed no anti-neuroinflammatory activity (>200 μM) in Table 3.

[0297] Table 3 a The maximum inhibitory concentration (IC 50 ) was calculated from the anti-neuritis activity of inhibiting LPS-induced nitric oxide (NO) in BV-2 cells. b The half inhibitory concentration (IC50) of CBD analogues for maximal inhibition of BV-2 cell viability 50 ). c The therapeutic index (TI) was determined by the following equation: TI = IC of cell viability 50 IC with / NO 50 . d Not measured.

[0298] Example 6

[0299] Structure-activity relationship of compounds 12a-12h

[0300] This example covers the effect of side chain functional groups on CBD activity. The side chains of CBD are crucial to its biological activity. The anti-neuroinflammatory activity of CBD derivatives 12a-12h, with the exception of 12e, significantly decreased with increasing linear alkane chain length (see Table 4). Furthermore, the therapeutic index of CBD derivatives 12a-12h showed similar trends.

[0301] Table 4 a The maximum inhibitory concentration (IC 50 ) was calculated from the anti-neuritis activity of inhibiting LPS-induced nitric oxide (NO) in BV-2 cells. b The half inhibitory concentration (IC50) of CBD analogues for maximal inhibition of BV-2 cell viability 50 ). c The therapeutic index (TI) was determined by the following equation: TI = IC of cell viability 50 IC with / NO 50 . d Not measured.

[0302] Example 7

[0303] Structure-activity relationship of compounds 19a-19d

[0304] Compound 12a, which has an ethyl group, exhibited the best therapeutic index among CBD derivatives 12a-12h. Therefore, this compound was further investigated for substitution of side-chain heterocyclic functional groups. Heterocyclic rings are key pharmacophores in many approved drugs for various therapeutic applications. Because thiazole-substituted side chains can enhance hydrophilicity, the question of whether the introduction of polar heterocyclic rings would lead to improved therapeutic efficacy was explored. Specifically, Example 4 outlines the introduction of a five-membered nitrogen heterocycle at the α-C position of compound 12a.

[0305] Among the four compounds tested, CIAC001, which introduced a 2H-1,2,3-triazole pharmacophore at the α-C position, exhibited the strongest anti-neuritis activity (2.5±0.7μM) and showed the best therapeutic index (Table 5). Compared with compound 12a, the anti-neuritis activity of CIAC001 was not significantly increased. However, the cytotoxicity of CIAC001 was reduced by half compared with compound 12a, indicating that the introduction of a 2H-1,2,3-triazole pharmacophore can improve cytotoxicity and increase the therapeutic window. Compound 19b, which introduced a 1H-1,2,3-triazole pharmacophore, showed an approximately 14.4-fold reduction in anti-neuritis activity compared with CIAC001.

[0306] Table 5 a The maximum inhibitory concentration (IC 50 ) was calculated from the anti-neuritis activity of inhibiting LPS-induced nitric oxide (NO) in BV-2 cells. b The half inhibitory concentration (IC50) of CBD analogues for maximal inhibition of BV-2 cell viability 50 ). c The therapeutic index (TI) was determined by the following equation: TI = IC50 of cell viability / IC50 of NO. d Not measured.

[0307] In summary, structure-activity relationship studies of the CBD side chain revealed that the CBD derivative CIAC001 exhibited a 4.9-fold increase in anti-neuroinflammatory activity and a 6.8-fold decrease in cytotoxicity compared to CBD. Furthermore, CIAC001's therapeutic index was increased by approximately 30-fold, extending its therapeutic window in in vitro cell studies and in vivo animal studies. Therefore, CIAC001 was selected as the lead compound as the most potent and least toxic CBD derivative.

[0308] Example 8

[0309] In vitro anti-inflammatory activity test of CIAC001

[0310] This example covers the effects of CIAC001, a representative compound disclosed herein, on microglial signaling. Activated microglia secrete proinflammatory factors such as IL-1β, TNF-α, and IL-6. CIAC001 dose-dependently inhibited LPS-induced overexpression of IL-1β (Figure 1A), TNF-α (Figure 1B), and IL-6 (Figure 1C). In contrast, CBD failed to inhibit LPS-induced overexpression of TNF-α and IL-6 mRNA, and only inhibited overexpression of IL-1β at 10 μM. In addition, it is worth noting that even at a concentration of 10 μM, CBD even enhanced LPS-induced IL-6 mRNA expression. This is not surprising, considering that 10 μM CBD may cause a toxic stress response.

[0311] Microglia change their morphology upon stimulation. Most untreated BV-2 cells have a small, round morphology. LPS activation leads to an increase in cell volume and a significant increase in the percentage of rod-shaped microglia, while CIAC001 significantly reverses the LPS-induced morphological changes (Figure 1D). These results indicate that CIAC001 inhibits microglial activation. In addition to cell morphology analysis, the effects of CIAC001 on microglial M1 and M2 polarization were further investigated. M1 microglia produce inflammatory mediators and induce neuroinflammation, while M2 microglia release anti-inflammatory factors and induce neuroprotection. The expression levels of M1 / M2 markers were measured at the mRNA level by qPCR. CIAC001 inhibited the LPS-induced overexpression of M1 phenotype markers IL-1β and iNOS, and alleviated the LPS-induced decrease in the expression of M2 phenotype marker IL-10 (Figure 7). Overall, these in vitro cell experiments consistently demonstrated that CIAC001 exhibits potent anti-neuroinflammatory activity.

[0312] Example 9

[0313] CIAC001 for the treatment of morphine addiction

[0314] This example covers the effects of CIACI001, a representative compound disclosed herein, on morphine addiction, withdrawal, and CPP, as well as the pharmacokinetics of CIAC001. Multiple studies have demonstrated that microglia play a neuromodulatory role in drug reward. Opioids activate microglia, releasing proinflammatory cytokines that affect reward circuits. This increases neuronal excitability and enhances mesolimbic dopamine signaling, thereby contributing to addiction.

[0315] The effects of compound CIAC001 on opioid withdrawal were investigated using naloxone-induced morphine withdrawal behavior (Figure 8A). Administration of naloxone to chronic morphine-addicted mice resulted in frequent jumping behavior. CIAC001 (2 μg / kg, 20 μg / kg, and 0.2 mg / kg, ip) dose-dependently reduced naloxone-induced withdrawal jumping in morphine-addicted mice, whereas CBD failed to significantly alleviate morphine withdrawal symptoms (Figure 2A). These results suggest that CIAC001 is effective for treating morphine withdrawal symptoms and preventing physical dependence.

[0316] Behavioral sensitization is a phenomenon in which motor activity increases over time after repeated exposure to psychostimulants and has become a useful model for evaluating the effects of pharmacological treatments for drug addiction. Therefore, a morphine-induced behavioral sensitization experiment was performed (Figure 8B). After repeated daily morphine exposure, mice showed an increased increase in movement distance (Figure 2B). CIAC001 (0.2 mg / kg, ip) significantly inhibited the development of morphine-induced behavioral sensitization during the induction period, while CBD (0.2 mg / kg, ip) had little effect in attenuating morphine-induced behavioral sensitization (Figure 2B). After a 1-week withdrawal period (days 8-14), mice were injected with morphine on day 15. Morphine injection significantly induced the expression of behavioral sensitization in morphine-pretreated mice (Figure 2C). CIAC001 (0.2 mg / kg, ip) reduced the increase in movement distance caused by morphine injection, but CBD (0.2 mg / kg, ip) had no effect (Figure 2C). These results indicate that CIAC001 inhibits the development and expression of morphine-induced behavioral sensitization.

[0317] Conditioned place preference (CPP, Figure 8C) is a standard preclinical behavioral model for measuring the rewarding effects of drugs, in which animals tend to show a preference for the environment paired with the drug (reflected by spending more time in it). Conditioning with morphine induced CPP in mice, and their CPP scores were significantly higher than those of the control group (Figure 2D). Administration of CIAC001 (0.2 mg / kg, ip) significantly reduced the expression of morphine-induced CPP (Figure 2D). After CPP behavioral testing on day 7, the experimental mice were sacrificed. Brain regions of the mPFC, NAc, and VTA involved in reward and addiction were dissected. As the most critical proinflammatory factor in neuroinflammation, the expression of IL-1β was measured. Chronic morphine treatment increased IL-1β expression in the mPFC (Figure 2E), NAc (Figure 2F), and VTA (Figure 2G) regions, while CIAC001 inhibited morphine-induced IL-1β overexpression in these regions (Figures 2E-G). Furthermore, administration of CIAC001 reduced the expression of the microglial activation marker Iba1 in the mPFC, but not in the NAc or VTA (Figure 9). These findings suggest that CIAC001 inhibits morphine-induced neuroinflammation and prevents morphine-induced CPP.

[0318] Given CIAC001's excellent activity in suppressing morphine addiction, it is likely that CIAC001 possesses strong blood-brain barrier penetration. CIAC001 and CBD were administered orally. Compound concentrations in plasma and brain were measured. Following oral administration (10 mg / kg, formulation: 5% DMSO / 10% Solutol / 85% Captisol (20%), po), CIAC001 and CBD peaked in the brain at 0.3 and 0.5 h, respectively.

[0319] Table 6 summarizes the pharmacokinetic parameters of CIAC001 and CBD in the brain of BLAB / c mice after oral administration of 10 mg / kg. max (175.0 ng / mg) is about twice that of CBD (98.1 ng / mg, Figure 10, Table 5). In addition, the half-life of CIAC001 (t 1 / 2 ) and mean residence time (MRT) were 0.8 and 0.9 h, respectively, which were higher than the t 1 / 2 (0.5h) and MRT (0.7h) (Table 6).

[0320] Table 6

[0321] Table 7 summarizes the brain-to-plasma (B / P) ratios of CIAC001 and CBD after oral administration as described above. Within the first 3 hours after administration, the B / P ratio of CIAC001 exceeded 200%, far exceeding the level of CBD (less than 100%, Table 7). The area under the curve (AUC) of CIAC001 in the brain (172.8 h ng / g) exceeded that of CBD (66.7 h ng / g) by more than two times. Compared with CBD, oral CIAC001 showed better blood-brain barrier permeability and central nervous system pharmacokinetics. The high blood-brain barrier permeability and central nervous system enrichment of CIAC001 may, at least in part, explain its lower levels in plasma (Figure 10).

[0322] Table 7 a The B / P concentration ratio was determined by the following equation: B / P = 100% x brain concentration / plasma concentration. b Not detected.

[0323] The combined results of these in vivo studies showed that CIAC001 alleviated morphine-induced withdrawal reactions and behavioral sensitization (CPP), which demonstrated the therapeutic effect of CIAC001 on morphine addiction.

[0324] Example 10

[0325] CIAC001 safety assessment

[0326] The present embodiment relates to the safety and reward effect evaluation of CIAC001. The safety analysis results of CIAC001 are shown in 3A-B. Respiratory depression was found after a single administration of THC (10 mg / kg, ip, Figure 3A) to mice. In contrast, CIAC001 (20 mg / kg, ip) did not affect respiratory rate (Figure 3A). In addition, a Y-type maze test was carried out to study the effect of THC or CIAC001 on spatial learning and memory in mice (Figure 8E). Compared with the control group, the time spent on the new arm by mice administered with THC (10 mg / kg, ip) was significantly reduced (Figure 3B). On the contrary, the time spent on the new arm by mice administered with CIAC001 (20 mg / kg, ip) was not affected (Figure 3B), indicating that it had no obvious memory decline or cognitive impairment.

[0327] To investigate the behavioral rewarding effects of CIAC001, a CPP experiment was conducted (Figure 8F). As shown in Figure 3C, mice treated with CIAC001 (20 mg / kg, ip) showed no effect on CPP after 6 days of training, suggesting a lack of rewarding effects of CIAC001. An open field test was conducted after seven consecutive days of CIAC001 (20 mg / kg, ip) administration (Figure 8D). There were no significant differences in the distance traveled (Figure 3D) or grooming and scratching behaviors (Figure 3E) of mice before and after CIAC001 (20 mg / kg, ip) treatment, indicating that CIAC001 did not cause stress in mice. Consistent with these observations, the body weight of CIAC001-treated mice remained unchanged during drug treatment compared to the control group (Figure 3F). Notably, the CIAC001 dose used in the in vivo safety assessment experiment (20 mg / kg) was 100 times the maximum dose used to prevent morphine addiction (0.2 mg / kg CIAC001). Therefore, compared to THC, CIAC001 is a safe and non-addictive synthetic cannabinoid.

[0328] Example 11

[0329] CIAC001 target protein fishing and PKM2 target validation

[0330] To identify the molecular target of CIAC001 in inhibiting neuroinflammation, a photoaffinity probe 19e was synthesized. The workflow for protein target identification is shown in Figure 4A. The photoaffinity probe 19e has a photoreactive diazo group and a terminal alkyne group. Under ultraviolet irradiation, the photoreactive group is activated and binds to protein macromolecules to form new covalent bonds. BV-2 cells were incubated with 19e (5 M) and different concentrations of CIAC001 (0 and 20 M) for 2 h and then irradiated with 365 nm UV light for 30 min. Subsequently, the cells were lysed, and biotin was attached to the probe using a copper-catalyzed alkyne-azide cycloaddition "click" reaction. The target protein sample was isolated using streptavidin magnetic beads. The samples were separated by SDS-PAGE and visualized by silver staining (Figure 4B). The band enriched by 19e was competitively inhibited by CIAC001 (20 M). In order to limit the modification effect on CIAC001, a non-covalent small molecule probe 19f without a photocrosslinking group was also used for target identification. Cell homogenate was incubated with 19f. The alkyne group of 19f and Biotin-PEG3-N3 were connected through a "click" reaction, and then affinity enriched using streptavidin magnetic beads (Figure 4A). The sample was separated by SDS-PAGE, and a similar spectrum to 19e was shown by silver staining. The bands captured by 19e or 19f were cut and then subjected to proteomic analysis by LC-MS / MS. The protein cross-target proteins captured by 19e and 19f were taken for subsequent protein target verification.

[0331] PKM (pyruvate kinase M1 / 2, approximately 58 kDa monomer), a representative binding protein of the CIAC001 probe (Figure 4C), is considered a key determinant in promoting inflammatory responses. Western blotting using the corresponding antibodies first verified whether PKM directly binds to CIAC001. As shown in Figure 5A, CIAC001 directly binds to PKM2, but not PKM1, both in situ and in vitro. Indeed, PKM2 expression is higher than PKM1 in BV2 microglial cells. Furthermore, 19e binding to PKM2 can be competitively inhibited by high concentrations of free CIAC001 (Figure 5A). Furthermore, a cell-based thermal stability assay (CETSA) was performed to confirm whether PKM2 is a binding target of CIAC001. Briefly, in a CETSA assay, cell lysates are incubated with CIAC001, heated to denature and precipitate proteins, and the denatured proteins are removed by centrifugation. Remaining soluble proteins are detected by quantitative Western blotting. Binding of CIAC001 increases the thermal stability of PKM2, as indicated by a decrease in its melting temperature (T m ) changes (ΔT m 5B). In contrast, 19b failed to affect the T of PKM2. m, which demonstrates the specificity of the interaction between CIAC001 and PKM2. To quantitatively characterize the interaction between CIAC001 and PKM2, a fluorescence titration experiment was performed on purified PKM2 protein using CIAC001 (Figure 5C). CIAC001 binding resulted in quenching of the intrinsic fluorescence of PKM2. The molar ratio of the CIAC001-PKM2 interaction was deduced to be 0.54±0.07 and the dissociation constant K D The binding of CIAC001 to PKM2 was 2.2±0.2μM (Figure 5C). Molecular docking simulations of CIAC001 binding to PKM2 revealed that CIAC001 binds to the interaction interface between two PKM2 monomers at a ratio of 1:2, consistent with the molar ratio of 0.54±0.07 measured by fluorescence titration (Figure 5D). CIAC001 binds tightly to PKM2 through hydrophobic interactions with surrounding residues of PKM2 (Figure 5E). Further molecular dynamics simulations showed that the binding of CIAC001 increased the number of hydrogen bonds between dimers in the tetrameric form (Figure 11). However, 19b binds within the monomer of PKM2, away from the dimer interface (Figures 5D, F).

[0332] The tetrameric form of PKM2 promotes ATP production through oxidative phosphorylation. CIAC001 administration increased ATP levels, while 19b showed no change (Figure 6A), suggesting that CIAC001 activates PKM2. Overall, these results indicate that PKM2 protein is a direct target of CIAC001. LPS treatment reduced the content of PKM2 tetramers in cells but increased the content of PKM2 monomers (Figure 6B). CIAC001 was able to reverse the LPS-induced decrease in PKM2 tetramers and inhibit the LPS-induced increase in PKM2 monomers (Figure 6B). Monomeric PKM2 translocates to the nucleus, where it acts as a transcriptional coactivator and enhances inflammatory responses. LPS treatment significantly increased the content of PKM2 in the nucleus (Figure 6C). CIAC001 completely reversed the LPS-induced nuclear PKM2 translocation (Figure 6C). Monomeric PKM2 can function as a glycolysis promoter and promote the Warburg effect. Exposure of microglial BV-2 cells to LPS increased lactate dehydrogenase A (LDHA) mRNA (Figure 12) and lactate content in the culture medium (Figure 6D). CIAC001 treatment significantly inhibited LPS-induced LDHA mRNA overexpression (Figure 12) and lactate release (Figure 6D), while 19b did not prevent the LPS-induced increase in lactate (Figure 6D), indicating that CIAC001 can reverse PKM2-mediated metabolic reprogramming.

[0333] Hif-1α is directly regulated by PKM2 and controls IL-1β expression. Long-term morphine treatment increased Hif-1α expression in the mPFC (Figure 6E). CIAC001 administration suppressed long-term morphine-induced Hif-1α overexpression (Figure 6E). These results indicate that CIAC001 treats morphine addiction through the PKM2-Hif-1α-IL-1β signaling pathway.

[0334] The primary targets of most cannabinoids are cannabinoid receptors (CB1 and CB2). To test the potential interaction of CIAC001 with CB1 / CB2, a cAMP assay for Gi-coupled GPCR signaling was performed. Unlike CBD, which has minimal CB1 / CB2 receptor activity, CIAC001 has no significant CB1 / CB2 activity (Figure 13, Table 8), ruling out CB1 / CB2 as a target of CIAC001. Overall, these results suggest that PKM2 is a specific target of CIAC001.

[0335] Table 8 a Not detected

[0336] (1'R,2'R)-4-(1-(2H-1,2,3-triazol-2-yl)ethyl)-5'-methyl-2'-(prop-1-en-2-yl)-6-(2-(3-(prop-2-yn-1-yl)-3H-diazirin-3-yl)ethoxy)-1',2',3',4'-tetrahydro-[1,1'-biphenyl]-2-ol(19e). CIAC001 (0.10 g, 0.3 mmol), Ph3P (0.09 g, 0.36 mmol, 1.2 equiv) and 2-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)ethan-1-ol (0.04 g, 0.36 mmol, 1.2 equiv) were dissolved in 10 mL of dry THF. Diisopropylazide dicarbonamide (0.20 g, 0.36 mmol) was dissolved in 2 mL of THF and added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with water, extracted with ether, and washed with salt water. The crude product was purified by column chromatography using petroleum ether / ethyl acetate. Get 19e: 34.19mg, 25.6%, 1H NMR (300MHz, CDCl3) δ=7.57 (s, 2H), 6.81 (d, J=2.3Hz, 2H), 5.79 (q, J=7.1Hz, 1H), 5.12 (s, 1H), 4.48 (s, 1H), 4.39 (s, 1 13C NMR (75MHz, CDCl3) δ=168.7, 149.9, 147.3, 139.6, 134.1, 133.2, 128.8, 12 8.8, 123.9, 111.3, 63.2, .45.4, 38.4, 30.3, 28.6, 23.4, 20.9, 20.8, 19.4. MS(ESI-)calcd for C27H33N5O2-H+:458.26[MH] - ;found:458.22.

[0337] (1'R,2'R)-4-(1-(2H-1,2,3-triazol-2-yl)ethyl)-5'-methyl-2'-(prop-1-en-2-yl)-6-(prop-2-yn-1-yloxy)-1',2',3',4'-tetrahydro-[1,1'-biphenyl]-2-ol (19f). CIAC001 (168.5 mg, 0.5 mmol) and 3-bromopropyne (0.6 mmol, 1.2 equiv) were dissolved in 15 mL of acetone, and K2CO3 (71.38 mg, 0.6 mmol, 1.2 equiv) was added to the mixture while stirring at room temperature. After 4 h, the reaction mixture was diluted with water and extracted with ethyl acetate, and the organic phase was washed with brine. The crude product was separated by column chromatography, eluting with petroleum ether / ethyl acetate. 19f: 51.87 mg, 27.5%, 1H NMR (300MHz, CDCl3) δ=7.64 (d, J=3.0Hz, 2H), 6.42 (d, J=13.5Hz, 1H), 6.23 (d, J =6.6Hz, 1H), 6.14 (s, 1H), 5.75 (q, J = 7.0Hz, 1H), 5.53 (s, 1H), 4.50 (s, 1H), 4.3 5(s, 1H), 4.02(s, 1H), 3.80–3.61(m, 2H), 2.41(s, 1H), 2.20(m, 2H), 2.07–1.99 (m, 5H), 1.96 (m, 3H), 1.85–1.75 (m, 6H), 1.63 (s, 3H), 1.27 (t, J=5.2Hz, 3H); 13C NMR (75MHz, CDCl3) δ=149.0, 134.0, 123.9, 117.6, 111.2, 100.0, 82.6, 69.3, 64.1, 62.9, 32.9, 32.3, 30.1, 26.5, 13.3. MS(ESI - )calcd for C23H27N3O2-H + :376.21[MH] - ;found:376.09.

[0338] Example 13

[0339] CIAC001 improves learning and memory abilities and alleviates anxiety in AD mice

[0340] This example covers the effects of CIAC001 on learning, memory, and anxiety. First, wild-type and transgenic (Alzheimer's disease model) mice were administered with either control or CIAC001 and subjected to the Morris water maze test. As shown in Figures 14A-14F, the escape latency of mice in the Morris water maze test 45 days after intraperitoneal injection of CIAC001 (14A), the escape latency on the fifth day of water maze training (14B), the exploration time of mice in the platform quadrant area (14C) and the number of times they crossed the platform (14D) on the first day after water maze training, and the exploration time of mice in the platform quadrant area (14E) and the number of times they crossed the platform (14F) on the third day after water maze training are respectively shown. Compared with the AD group, the escape latency of AD mice in the WT group, 0.2 mg / kg CIAC001, 2 mg / kg CIAC001 and 20 mg / kg CIAC001-administered AD mice was reduced, and in the short-term and long-term memory tests after training, the exploration time and the number of times they crossed the platform quadrant in AD mice administered CIAC001 in each group increased, indicating that CIAC001 treatment can improve the learning and memory ability of mice.

[0341] Mice were subjected to a small open field test 45 days after intraperitoneal injection of CIAC001. The experimental results are shown in Figures 14G-14I. The number of grid crossings (14G), the number of standing times (14H), and the number of excretions (14I) in the small open field test showed that AD mice had significantly fewer grid crossings and standing times than WT mice, while the number of excretions was significantly increased. After treatment with CIAC001, the number of grid crossings and standing times in the small open field in the treated group of AD mice increased significantly, while the number of excretions decreased significantly, indicating that CIAC001 treatment can enhance the exploratory ability of AD mice and improve their anxiety.

[0342] Mice were subjected to a novel object recognition test 45 days after intraperitoneal injection of CIAC001. As shown in Figure 14J, compared with the AD group, AD mice treated with 20 mg / kg CIAC001 showed a significant increase in the novel object recognition index, indicating that CIAC001 treatment can enhance the cognitive and exploratory abilities of AD mice.

[0343] Mice were subjected to a Y-maze test 45 days after intraperitoneal injection of CIAC001. Figures 14K-14L show the total number of alternations (14K) and the effective alternation rate (14L) in the Y-maze test 45 days after intraperitoneal injection of CIAC001. The total number of alternations and the effective alternation rate in AD mice were significantly reduced compared to WT mice. However, after CIAC001 treatment, the total number of alternations and the effective alternation rate in AD mice increased significantly, indicating that CIAC001 treatment can enhance the mice's exploratory ability and alleviate their anxiety.

[0344] Mice were subjected to the elevated plus maze test 45 days after intraperitoneal injection of CIAC001. Figures 14M-14N show, respectively, the number of open arm entries (14M) and time spent (14N) in the elevated plus maze test 45 days after intraperitoneal injection of CIAC001. In the elevated plus maze, the number of open arm entries and time spent in the elevated plus maze were significantly increased in the WT group, as well as in AD mice treated with 2mg / kg CIAC001 and 20mg / kg CIAC001, compared to the AD group. This suggests that CIAC001 treatment enhances the exploratory ability and reduces anxiety in AD mice.

[0345] Example 14

[0346] CIAC001's mechanism of action in treating AD

[0347] To investigate the mechanism of action of CIAC001 in the treatment of Alzheimer's disease, the transgenic mice were subjected to in vitro hippocampal potential measurements. The results of these analyses (Figures 15A-B) showed that CIAC001 increased hippocampal potential by nearly 500%.

[0348] Transgenic mice that received 0, 0.2, 2, or 20 mg / kg CIAC001 were immunoblotted for multiple Alzheimer's disease markers (Figure 15C-D). These analyses showed that CIAC001 inhibited the phosphorylation of Tau and TrkB and reduced the levels of Syn1 and Aβ.

[0349] Although the present invention has been described in its presently preferred embodiments, it will be appreciated that various modifications can be made without departing from the spirit of the invention. Accordingly, the invention is not to be limited except as by the claims that follow.

Claims

1. A method for treating or preventing opioid addiction, withdrawal symptoms or overdose, comprising administering to a subject a compound of formula I: or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Select from the following structures: -H， or R 2A and R 2B are independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C 4， Where R 2A and R 2B You can choose diaziridine instead; R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2 and Subscript n is 0 to 12; Thereby treating or preventing opioid addiction, withdrawal, or overdose.

2. According to claim 1, the compound of formula I may preferably be a compound of formula Ia:

3. According to claim 1 R 1 It can be preferably selected from the following structures: H, or 4. According to claim 3R 1 It can be preferably 5. According to claim 1 R 2A and R 2B Each is independently preferably selected from the following structures: -H or C1-C4 alkane.

6. According to claim 5 R 2A and R 2B Each is independently preferably selected from the following structures: -H or -CH3.

7. According to claim 6 R 2A and R 2B It may preferably be -H.

8. According to claim 1 R 3A and R 3B Each is independently preferably selected from the following structures: -H, -Cl, -Br, or -COOH.

9. According to the claim R 3A and R 3B It may preferably be -H.

10. According to claim 1, n can preferably be 0 or 1.

11. According to claim 1, the structure of formula I can be preferably 12. The method of claim 1, wherein the method comprises treating or preventing opioid addiction.

13. According to claim 1, wherein the method comprises treating or preventing opioid withdrawal symptoms.

14. According to claim 1, the dose of Compound I is about 0.5 μg / kg, about 2 μg / kg, about 20 μg / kg, about 200 μg / kg, about 2 mg / kg, about 20 mg / kg or about 100 mg / kg.

15. According to claim 1, the C of compound I in the brain of the subject MAX About 1-1000ng / mg.

16. According to claim 1, the plasma ratio of Compound I in the brain of the subject within 1 hour after administration is about at least 10:1, at least 25:1, at least 50:1, at least 100:1 or at least 200:

1.

17. According to claim 1, wherein the opioid is fentanyl, ibuprofen, codeine, dextromorphine, dihydrocodeine, enkephalin, heroin, oxycodone, oxymorphine, pethidine, methadone, morphine, nikomophine, opium, codeine oxide, hydroxymorphine, perphenazine, perphenaline, a derivative thereof, a precursor thereof, or a pharmacologically acceptable salt or solvate thereof.

18. The method of claim 17, wherein the opioid is morphine.

19. The subject according to claim 1 , exhibits symptoms of opioid addiction, including withdrawal symptoms, behavioral sensitization, or conditioned place preference.

20. According to claim 1, Compound I can be administered orally, subbuccally, sublingually, rectally, vaginally, intravenously, intraarterially, intramedullary, intramuscularly, intracerebrally, intraventricularly, intraspinally, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, or via a mucosal oral route, or by inhaler.

21. According to claim 1, Compound I can be administered together with a pharmacologically acceptable excipient.

22. The method of claim 1, further comprising administering to the subject an opioid antagonist.

23. According to claim 22, the opioid antagonist is naloxone or naltrexone.

24. According to claim 1, the subject is addicted to opioids.

25. According to claim 1, the dose of Compound I is sufficient to inhibit the overexpression of IL-1β, TNFα, IL-6 induced by opioids in a subject.

26. According to claim 1, a sufficient dose of Compound I can modulate the expression of PKM, heat shock protein-60, alpha-enolase, elongation factor 1 alpha, RNA helicase, 90 kilodalton heat shock protein, ubiquitin carboxyl terminal hydrolase, T-complex protein 1 subunit alpha, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor family B member 6a or elongation factor 1 gamma in a subject.

27. According to claim 26, a sufficient dose of Compound I can modulate PKM in a subject.

28. According to claim 27, wherein the PKM is PKM2.

29. According to claim 1, the dose of Compound I is sufficient to increase the ratio of M2 to M1 microglia in the brain of the subject.

30. According to claim 1, Compound I does not induce a toxic stress response in the subject.

31. A method of regulating PKM, heat shock protein-60, alpha-enolase, elongation factor 1 alpha, RNA helicase, 90 kilodalton heat shock protein, ubiquitin carboxyl terminal hydrolase, T-complex protein 1 subunit alpha, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor B family member 6a, elongation factor 1 gamma in a subject, comprising administering compound I to a subject in need thereof: or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures: -H, or R 2A and R 2B are independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C 4， Where R 2A and R 2B You can choose diaziridine instead; R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and Subscript n is 0 to 12; It can thereby regulate PKM, heat shock protein-60, α-enolase, elongation factor 1α, RNA helicase, heat shock protein 90β, ubiquitin carboxyl-terminal hydrolase, T complex protein α subunit, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor clade B member 6a, and elongation factor 1γ in the subject's body.

32. According to claim 31, wherein the method comprises modulating PKM in the subject.

33. According to claim 32, wherein the PKM is PKM2.

34. A method for increasing the ratio of M2 to M1 microglia in the brain of a subject, comprising administering to the subject a compound I: or a salt or solvate thereof, wherein the main body of the compound comprises: R 1 Selected from the following structures: -H, or R 2A and R 2B are independently selected from -H, C1-C4 alkanes, C1-C4 alkenes, or C1-C 4， Where R 2A and R 2B You can choose to use diaziridine instead; R 3A and R 3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and Subscript n is 0 to 12; This increased the ratio of M2 to M1 microglia in the subjects' brains.

35. A method of treating a subject suffering from a neurodegenerative disease or disorder, comprising administering thereto Compound I: or a salt or solvate thereof, wherein the main body of the compound comprises: R1 is selected from the following structures: -H， or R 2A and R 2B are independently selected from the following structures: -H, C1-C4 alkane, C1-C4 alkene, or C1-C4, wherein R 2A and R 2B You can choose to use diaziridine instead; R3A and R3B are independently selected from the following structures: -H, halogen, -COOH, or -NH2; and Subscript n is 0 to 12; Thereby treating neurodegenerative diseases such as Alzheimer's disease.

36. According to claim 35, the compound of formula I may preferably be a compound of formula Ia:

37. According to claim 35, R 1 Selected from the following structures: H, or 38. According to claim 35, R 1 for 39. According to claim 35, R 2A and R 2B Each is independently preferably selected from the following structures: -H or C1-C4 alkane.

40. According to claim 39, R 2A and R 2B Each is independently preferably selected from the following structures: -H or -CH3.

41. According to claim 40, R 2A and R 2B It may preferably be -H.

42. According to claim 35, R 3A and R 3B Each is independently preferably selected from the following structures: -H, -Cl, -Br or -COOH.

43. According to claim 42, R 3A and R 3B It may preferably be -H.

44. According to claim 35, n is preferably 0 or 1.

45. According to claim 35, the structure of formula I is preferably 46. ​​According to claim 35, the disease or disorder may be selected from Alzheimer's disease, Parkinson's disease or amyotrophic lateral sclerosis.

47. According to claim 35, Compound I inhibits neuroinflammation in a subject.

48. According to claim 35, compound I has an IC of 50 , which is at most about 200 μM, 150 μM, 100 μM, 75 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 2.5 μM or 1 μM.

49. According to claim 35, the cytotoxic IC50 of Compound I in microglia is at least about 1 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 75 μM, 100 μM, 150 μM, 200 μM.

50. According to claim 35, the dose of Compound I is sufficient to reduce the expression of IL-1β in microglia in the subject by at least 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold or 20-fold.

51. According to claim 35, the dose of Compound I is sufficient to reduce the expression of TNFα in microglia in a subject by at least 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold or 20-fold.

52. According to claim 35, the dose of Compound I is sufficient to reduce the expression of IL-6 in microglia in a subject by at least 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 6-fold, 8-fold, 10-fold, 15-fold or 20-fold.

53. According to claim 35, the dose of Compound I is about 0.5 μg / kg, 2 μg / kg, 20 μg / kg, 200 μg / kg, 2 mg / kg, 20 mg / kg or 100 mg / kg.

54. According to claim 35, the C of Compound I in the brain of a subject MAX About 1-1000ng / mg.

55. According to claim 35, the brain to plasma ratio of Compound I within 1 hour after administration is at least about 10:1, 25:1, 50:1, 100:1 or 200:

1.

56. According to claim 35, Compound I can be administered orally, subbuccally, sublingually, rectally, vaginally, intravenously, intraarterially, intramedullary, intramuscularly, intracerebrally, intraventricularly, intraspinally, subcutaneously, intraperitoneally, intraocularly, intranasally, transdermally, epidurally, intracranially, transdermally, intrauterinely, intravitreally, or via a mucosal oral route, or by inhaler.

57. According to claim 35, Compound I may be administered together with a pharmacologically acceptable excipient.

58. According to claim 35, a sufficient dose of Compound I can modulate the dose of PKM, heat shock protein-60, alpha-enolase, elongation factor 1 alpha, RNA helicase, 90 kilodalton heat shock protein, ubiquitin carboxyl terminal hydrolase, T-complex protein 1 subunit alpha, phosphoglycerate kinase 1, elongation factor Tu serine, elongation factor Tu cysteine, peptidase inhibitor family B member 6a, or elongation factor 1 gamma in a subject.

59. According to claim 58, a sufficient dose of Compound I can modulate PKM in a subject.

60. According to claim 59, wherein the PKM is PKM2.

61. According to claim 35, the dose of Compound I is sufficient to increase the ratio of M2 to M1 microglia in the brain of the subject.

62. According to claim 35, Compound I does not induce a toxic stress response in the subject.