A compound for preventing and treating pain and its application

Abstract

The present invention discloses an agonist of a TREK-1 potassium ion channel and its application in analgesic therapy. Specifically, the present invention discloses a compound that can activate the TREK-1 potassium ion channel, the compound has a significant analgesic effect, and can be used to prepare a pharmaceutical composition and preparation for preventing and / or treating chronic pain (muscle and soft tissue pain, bone and joint pain, headache, visceral pain, pathological neuralgia), acute pain, and cancer pain.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and specifically relates to a compound for preventing and treating pain and application thereof. Background Art

[0002] Pain causes patients to suffer long-term torture and even produce malignant emotional reactions such as anxiety and depression. At present, the clinical drugs used to treat pain are mainly opioid analgesics, non-steroidal anti-inflammatory drugs, anticonvulsants, depressants and other drugs. Opioids have good analgesic effects, but these drugs have toxic side effects such as addiction and respiratory depression, which bring great harm. Non-steroidal anti-inflammatory drugs have weak efficacy and strong gastrointestinal and cardiovascular side effects. Pregabalin is a drug for the treatment of neuropathic pain and herpes zoster, but the treatment effect is not ideal, and only half of the patients who take the drug get partial relief. Therefore, the development of new drugs with new mechanisms is an urgent need for the treatment of pain.

[0003] TREK-1 potassium channel belongs to the family of two-pore potassium ion channels, mediates background potassium current, and participates in regulating nerve excitability. After knocking out the corresponding gene of TREK-1, the pain sensitivity of rodents to thermal pain and mechanical pain increased (Alloui et al. EMBO J. 2006, 25: 2368-2376). TREK-1 plays an important role in the morphine analgesic pathway and is not related to the addiction, respiratory depression and constipation effects of morphine (Devilliers et al. Nat Commun. 2013, 4: 2941). Therefore, activating the TREK-1 channel is a new therapeutic strategy for treating pain such as neuralgia and inflammatory pain, and finding agonists of the TREK-1 channel is a new direction for developing new analgesic drugs. Summary of the invention

[0004] In order to solve the deficiencies in the prior art, the object of the present invention is to provide a TREK-1 agonist and its application in treating pain.

[0005] The present invention provides a use of a compound (YET01) represented by formula (I) or a pharmaceutically acceptable salt thereof, or a compound (YET02) represented by formula (II) or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition or preparation for preventing and / or treating pain.

[0006]

[0007] In another preferred embodiment, the compound of formula (I) or (II) has agonist activity on the TREK-1 channel.

[0008] In another preferred embodiment, the compound of formula (I) is compound YET01, and the two are the same compound.

[0009] In another preferred embodiment, the compound of formula (II) is compound YET02, and the two are the same compound.

[0010] In another preferred embodiment, the pain includes chronic pain, acute pain, cancer pain, etc.

[0011] In another preferred embodiment, the chronic pain includes muscle and soft tissue pain, bone and joint pain, headache, visceral pain, pathological neuralgia, etc.

[0012] In another preferred embodiment, the muscle and soft tissue pain includes one or more of myofasciitis, tenosynovitis, periarthritis of the shoulder, muscle strain pain, fibromyalgia, cold pain, burn pain, toothache, etc.

[0013] In another preferred embodiment, the bone and joint pain is one or more of knee pain, ankle pain, wrist pain, elbow pain, shoulder pain, patellar pain, hip pain, femoral pain, ankylosing spondylitis, sacroiliitis, rheumatoid arthritis, rheumatoid arthritis, gouty arthritis, intervertebral disc herniation, cervical pain, lumbar pain, etc.

[0014] In another preferred embodiment, the headache includes primary headache, secondary headache, cranial neuralgia, central and primary facial pain and other headaches.

[0015] In another preferred embodiment, the primary headache includes one or more of migraine without aura, migraine with aura, hemiplegic migraine, chronic migraine, migraine complications, paroxysmal syndromes associated with migraine, tension-type headache, trigeminal autonomic headache and other primary headaches.

[0016] In another preferred embodiment, the secondary headache includes one or more of headache due to head and neck trauma, headache due to non-vascular intracranial disease, headache due to substances or substance withdrawal, headache due to head and neck vascular disease, headache due to internal environment disorder, headache due to mental illness, headache and facial pain due to lesions of the head, neck, eyes, ears, nose, sinuses, teeth, mouth or other head and facial structures.

[0017] In another preferred embodiment, the cranial neuralgia, central and primary facial pain and other headaches include one or more of trigeminal neuralgia, glossopharyngeal neuralgia, intermedius neuralgia (facial neuralgia), occipital neuralgia, optic neuritis, etc.

[0018] In another preferred embodiment, the visceral pain includes pain from visceral organs, including the respiratory tract, gastrointestinal tract, pancreas, urethra, kidney, gallbladder, bladder and genitals.

[0019] In another preferred embodiment, the pathological neuralgia includes post-herpetic neuralgia, diabetic neuropathy, painful HIV-related sensory neuropathy, burning syndrome, post-amputation pain, phantom pain, painful neuroma; traumatic neuroma, nerve compression injury, spinal stenosis, carpal tunnel syndrome, radicular pain, sciatica; nerve avulsion injury, brachial plexus avulsion injury; complex regional pain syndrome, neuralgia caused by drug therapy, neuralgia caused by cancer chemotherapy; pain after spinal cord injury, primary small fiber neuropathy, primary sensory neuropathy, trigeminal autonomic headache, etc. One or more of the above.

[0020] In another preferred embodiment, the acute pain includes one or more of acute traumatic pain, postoperative pain, labor pain, visceral pain, toxic and thermal pain, itching, postoperative pain, etc.

[0021] In another preferred embodiment, the cancer in the cancer pain includes adenocarcinoma in glandular tissue, germ cell tumor in organ embryonic tissue, cancer in epithelial tissue, leukemia in blood cell-forming tissue, lymphoma in lymphoid tissue, myeloma in bone marrow, sarcoma in connective tissue or supporting tissue, adrenal cancer, AIDS-related lymphoma, bladder cancer, bone cancer, brain cancer, breast cancer, carcinoid tumor, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, gastric cancer, head cancer, neck cancer, hepatobiliary cancer, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, Hodgkin's disease, non-Hodgkin's disease, nervous system tumors, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, skin cancer, gastric cancer, testicular cancer, thyroid cancer, urethral cancer, bone marrow cancer, multiple myeloma, tumors metastasized to bones, tumors infiltrating nerves and hollow organs, tumors near neural structures, etc. One or more of the above.

[0022] In another preferred embodiment, the dosage of the compound of formula (I) is 0.1-50 mg per kilogram of individual body weight (0.1-50 mg / kg); preferably, 5-20 mg / kg.

[0023] In another preferred embodiment, the dosage of the compound of formula (II) is 0.1-50 mg per kilogram of individual body weight (0.1-50 mg / kg); preferably, 5-20 mg / kg.

[0024] In another preferred embodiment, when the administration dose of the compound of formula (I) is ≤50 mg / kg, there is no adverse reaction.

[0025] In another preferred embodiment, the pharmaceutical composition further contains other active ingredients for treating pain.

[0026] Among them, the active ingredients include but are not limited to one or more of opioid analgesics, non-opioid analgesics, fluoxetine, etc., the opioid analgesics include morphine, heroin, etc.; the non-opioid analgesics include but are not limited to one or more of paracetamol, acetylsalicylic acid, ibuprofen, etocrylene, fenbufen, fenoprofen, ketorolac, flurbiprofen, indomethacin, ketoprofen, naproxen, etc., and their pharmaceutically acceptable salts, optically active racemates and mixtures.

[0027] In another preferred embodiment, the pharmaceutical composition contains 0.001-99 wt %; preferably, 0.1-90 wt %; further preferably, 1-80 wt % of the compound of formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof, based on the total weight of the composition.

[0028] In another preferred embodiment, the pharmaceutical composition is in the form of an oral dosage form or an injection dosage form.

[0029] In another preferred embodiment, the oral dosage forms include tablets, capsules, films, granules, etc., and also include sustained-release or non-sustained-release dosage forms.

[0030] The present invention also provides a pharmaceutical composition, comprising a compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a compound represented by formula (II) or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is as follows:

[0031]

[0032] In another preferred embodiment, the pharmaceutical composition further contains a pharmaceutically acceptable carrier.

[0033] In another preferred embodiment, the total content of the compound of formula (I) and / or the compound of formula (II) or a pharmaceutically acceptable salt thereof is 0.001-99 wt % of the composition; preferably, 0.1-90 wt %; further preferably, 1-80 wt %.

[0034] In another preferred embodiment, the pharmaceutical composition is used to prevent and / or treat pain.

[0035] In another preferred embodiment, the pain includes chronic pain (muscle and soft tissue pain, bone and joint pain, headache, visceral pain, pathological neuralgia, etc.), acute pain, cancer pain, etc.

[0036] In another preferred embodiment, the muscle and soft tissue pain includes one or more of myofasciitis, tenosynovitis, periarthritis of the shoulder, muscle strain pain, fibromyalgia, cold pain, burn pain, toothache, etc.

[0037] In another preferred embodiment, the bone and joint pain is one or more of knee pain, ankle pain, wrist pain, elbow pain, shoulder pain, patellar pain, hip pain, femoral pain, ankylosing spondylitis, sacroiliitis, rheumatoid arthritis, rheumatoid arthritis, gouty arthritis, intervertebral disc herniation, cervical pain, lumbar pain, etc.

[0038] In another preferred embodiment, the headache includes primary headache, secondary headache, cranial neuralgia, central and primary facial pain and other headaches.

[0039] In another preferred embodiment, the primary headache includes one or more of migraine without aura, migraine with aura, hemiplegic migraine, chronic migraine, migraine complications, paroxysmal syndromes associated with migraine, tension-type headache, trigeminal autonomic headache and other primary headaches.

[0040] In another preferred embodiment, the secondary headache includes one or more of headache due to head and neck trauma, headache due to non-vascular intracranial disease, headache due to substances or substance withdrawal, headache due to head and neck vascular disease, headache due to internal environment disorder, headache due to mental illness, headache and facial pain due to lesions of the head, neck, eyes, ears, nose, sinuses, teeth, mouth or other head and facial structures.

[0041] In another preferred embodiment, the cranial neuralgia, central and primary facial pain and other headaches include one or more of trigeminal neuralgia, glossopharyngeal neuralgia, intermedius neuralgia (facial neuralgia), occipital neuralgia, optic neuritis, etc.

[0042] In another preferred embodiment, the visceral pain includes pain from visceral organs, including the respiratory tract, gastrointestinal tract, pancreas, urethra, kidney, gallbladder, bladder and genitals.

[0043] In another preferred embodiment, the pathological neuralgia includes post-herpetic neuralgia, diabetic neuropathy, painful HIV-related sensory neuropathy, burning syndrome, post-amputation pain, phantom pain, painful neuroma; traumatic neuroma, nerve compression injury, spinal stenosis, carpal tunnel syndrome, radicular pain, sciatica; nerve avulsion injury, brachial plexus avulsion injury; complex regional pain syndrome, neuralgia caused by drug therapy, neuralgia caused by cancer chemotherapy; pain after spinal cord injury, primary small fiber neuropathy, primary sensory neuropathy, trigeminal autonomic headache, etc. One or more of the above.

[0044] In another preferred embodiment, the acute pain includes one or more of acute traumatic pain, postoperative pain, labor pain, visceral pain, toxic and thermal pain, itching, postoperative pain, etc.

[0045] In another preferred embodiment, the cancer in the cancer pain includes adenocarcinoma in glandular tissue, germ cell tumor in organ embryonic tissue, cancer in epithelial tissue, leukemia in blood cell-forming tissue, lymphoma in lymphoid tissue, myeloma in bone marrow, sarcoma in connective tissue or supporting tissue, adrenal cancer, AIDS-related lymphoma, bladder cancer, bone cancer, brain cancer, breast cancer, carcinoid tumor, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, gastric cancer, head cancer, neck cancer, hepatobiliary cancer, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, Hodgkin's disease, non-Hodgkin's disease, nervous system tumors, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, skin cancer, gastric cancer, testicular cancer, thyroid cancer, urethral cancer, bone marrow cancer, multiple myeloma, tumors metastasized to bones, tumors infiltrating nerves and hollow organs, tumors near neural structures, etc. One or more of the above.

[0046] In another preferred embodiment, the pharmaceutical composition can also be used in combination with other drugs.

[0047] In another preferred embodiment, the other drugs include one or more of fluoxetine, opioid analgesics, and non-opioid analgesics; the opioid analgesics include morphine, heroin, etc.; the non-opioid analgesics include but are not limited to one or more of paracetamol, acetylsalicylic acid, ibuprofen, etocrylene, fenbufen, fenoprofen, ketorolac, flurbiprofen, indomethacin, ketoprofen, naproxen, etc., and pharmaceutically acceptable salts, optically active racemates and mixtures thereof.

[0048] In another preferred embodiment, the pharmaceutical composition can be prepared into an oral dosage form or an injection dosage form;

[0049] The oral dosage forms include tablets, capsules, films, granules and the like.

[0050] In another preferred embodiment, the pharmaceutical composition can be administered to the desired subject (such as human and non-human mammals) by oral administration, injection, aerosol inhalation, etc.

[0051] The present invention also provides a method for preventing and / or treating pain, comprising:

[0052] The compound of formula (I) or the compound represented by formula (II) or a pharmaceutically acceptable salt or pharmaceutical composition thereof as described above is administered to a subject in need thereof.

[0053] In another preferred embodiment, the subject includes humans and non-human mammals (such as rodents and primates).

[0054] In another preferred embodiment, the subject is a human, and the dosage of the compound of formula (I) and / or the compound of formula (II) is 1 to 20 mg / kg; preferably, 5 mg / kg.

[0055] In another preferred embodiment, the frequency of administration is 1-4 times / day; preferably, 1 time / day.

[0056] In another preferred embodiment, the administration period is 2 weeks to 5 years; preferably, 2 months to 1 year.

[0057] In another preferred embodiment, the administration method is oral, injection, aerosol inhalation, etc.; the administration method is selected according to actual needs.

[0058] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0059] The beneficial effects of the present invention are:

[0060] (a) YET01 has good TREK-1 agonist activity;

[0061] (b) YET01 has good analgesic effect in multiple analgesic models. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 For virtual screening of YET01( Figure 1 A), YET02( Figure 1 B) Schematic diagram of their respective binding to TREK-1 channels.

[0063] Figure 2 Schematic diagram of YET01's ability to activate TREK-1; Figure 2 A is a schematic diagram of a representative current curve of HEK203-T cells expressing TREK-1 channels activated by 10 μM YET01 (the curves from top to bottom are: YET01, elution, and before administration). Figure 2 B is a schematic diagram of the concentration-dependence curve of YET01 activating TREK-1 channels.

[0064] Figure 3 Schematic diagram of YET02's ability to activate TREK-1; Figure 3 A is a schematic diagram of a representative current curve of HEK203-T cells expressing TREK-1 channels activated by 10 μM YET02 (the curves from top to bottom are: YET02, elution, and before administration). Figure 3B is a schematic diagram of the concentration-dependence curve of YET02 activating TREK-1 channels.

[0065] Figure 4 Schematic diagram of the analgesic ability of compound YET01 in the hot water tail-flick experiment in mice (the curves from top to bottom are: YET01, Vehicle).

[0066] Figure 5 Schematic diagram of the analgesic ability of compound YET01 in the cold water tail flick experiment in mice (the curves from top to bottom are: YET01, Vehicle).

[0067] Figure 6 Schematic diagram of the analgesic ability of compound YET01 in the CFA-induced chronic inflammatory pain model experiment in mice (the curves from top to bottom are: YET01, Vehicle).

[0068] Figure 7 Schematic diagram showing the analgesic ability of compound YET01 in the spontaneous pain behavior experiment in the mouse preserved nerve injury model.

[0069] Figure 8 Schematic diagram of the analgesic ability of compound YET01 in the cold plate experiment of the mouse retained nerve injury model (the upper left of the curve is: Pregabalin; the right side from top to bottom are: YET01 50 mg / kg, po., YET01 20 mg / kg, po., Vehicle). DETAILED DESCRIPTION

[0070] The inventors have conducted extensive and in-depth research and discovered a compound that activates the TREK-1 potassium channel by combining computer-aided drug design with electrophysiological testing. Experiments have shown that the compound of the present invention, as an agonist of the TREK-1 channel, has a significant analgesic effect and can be used to prevent and / or treat pain. On this basis, the present invention was completed.

[0071] Specifically, the present invention designs drugs for the pocket on the extracellular side composed of the P1 loop and the TM4 helix, and performs activity tests through whole-cell electrophysiological detection. It is found that compounds YET01 and YET02 have a significant effect of activating TREK-1 channels, and therefore can be used as TREK-1 agonists.

[0072] The present invention uses C57 mice to conduct behavioral efficacy experiments on pain resistance, and uses different doses of YET01 and YET02 via oral administration. The results show that compounds YET01 and YET02 have obvious analgesic effects, and therefore can be used for the preparation of drugs for treating pain.

[0073] Compounds of the present invention

[0074] As used in the present invention, the terms "compound of the present invention", "compound of formula (I)", "compound YET01", or "YET01" are used interchangeably to refer to the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0075] As used in the present invention, the terms "compound of formula (II)", "compound YET02", or "YET02" are used interchangeably to refer to the compound represented by formula (II) or a pharmaceutically acceptable salt thereof.

[0076] The present invention also includes pharmaceutically acceptable salts of the compounds of formula (I) and / or (II). The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention formed with acids or bases that are suitable for use as medicines. Pharmaceutically acceptable salts include inorganic salts or organic salts.

[0077] Compositions and methods of administration

[0078] The present invention provides a pharmaceutical composition for preventing and / or treating pain, which contains the above-mentioned compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof.

[0079] The pharmaceutical composition of the present invention can be used to prevent and / or treat pain. When using the pharmaceutical preparation of the present invention, other pain treatment agents such as fluoxetine, opioid analgesics, non-opioid analgesics, etc. can also be used simultaneously.

[0080] The pharmaceutical composition of the present invention contains a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable carrier or excipient. Such carriers include (but are not limited to): one or more of saline, buffer, glucose, water, glycerol, ethanol, powder, etc. The pharmaceutical preparation should match the administration method.

[0081] The pharmaceutical composition of the present invention can be prepared in the form of an injection, for example, by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably manufactured under aseptic conditions. The pharmaceutical composition of the present invention can also be prepared in powder form for aerosol inhalation.

[0082] The dosage of the active ingredient is a therapeutically effective amount, for example, about 1 μg / kg body weight to about 50 mg / kg body weight per day; preferably, about 5 μg / kg body weight to about 10 mg / kg body weight; further preferably, about 10 μg / kg body weight to about 5 mg / kg body weight. In addition, the compounds of the present invention can also be used together with other therapeutic agents.

[0083] The pharmaceutical composition of the present invention can be administered to a desired subject (such as a human or non-human mammal) in a conventional manner, and representative administration methods include (but are not limited to): oral administration, injection, aerosol inhalation, and the like.

[0084] When using a pharmaceutical composition, a safe and effective amount of the drug is administered to a mammal, wherein the safe and effective amount is usually at least about 10 micrograms / kg body weight, and in most cases does not exceed about 50 milligrams / kg body weight, preferably, the dose is about 10 micrograms / kg body weight to about 20 milligrams / kg body weight. Of course, the specific dose should also take into account factors such as the route of administration and the patient's health status, which are all within the skill of a skilled physician.

[0085] The present invention is further described below in conjunction with specific examples. The process, conditions, reagents, experimental methods, etc. for implementing the present invention, except for the contents specifically mentioned below, are all common knowledge and common common sense in the art, and the present invention has no particular restrictions. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for which specific conditions are not indicated in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0086] Example 1

[0087] Whole-cell electrophysiological assays

[0088] The cDNA of human TREK-1 was subcloned into the pEGFP-N1 expression vector. The constructed plasmid was transfected into CHO-K1 cells using Lipofectamine 3000. The cell culture medium used in the experiment was DMEM / F12, containing 10% FBS and 1% penicillin-streptomycin. The culture conditions were a constant temperature of 37°C and a 5% CO2 gas environment.

[0089] Cells cultured for 16-36 hours after transfection were used for electrophysiological experiments. Whole-Cell Recording was used for whole-cell patch clamp recording. The experiments were performed using a HEKA EPC10 amplifier, a Sutter Instrument MP-225 micromanipulator, a Nikon fluorescence microscope, and a Bio-Logic RSC-200 drug delivery system. Glass electrodes were pulled using a Sutter Instrument P-97 electrode puller with an inlet resistance of 3-7 MΩ. The intracellular solution of TREK-1 cells contained 140 mM KCl, 2 mM MgCl2, 10 mM EGTA, 1 mM CaCl2, 10 mM HEPES, and the pH was adjusted to 7.3 with KOH; the extracellular solution contained 150 mM NaCl, 5 mM KCl, 0.5 mM CaCl2, 1.2 mM MgCl2, 10 mM HEPES, and the pH was adjusted to 7.3 with NaOH. The IV curve of TREK-1 was detected using a ramp from -100mV to +100mV with a duration of 1s, and the clamping voltage was -80mV. The detected current signal was filtered using a low-frequency filter with a frequency of 2kHz, and the sampling frequency was 10kHz.

[0090] like Figure 2 As shown, compound YET01 has a significant ability to activate TREK-1 channels and is an agonist of TREK-1.

[0091] like Figure 3 As shown, compound YET02 has a significant ability to activate TREK-1 channels and is an agonist of TREK-1.

[0092] Example 2

[0093] Animal pharmacology and efficacy experiments

[0094] The test drugs used in this embodiment include:

[0095] YET01 was synthesized by conventional methods, and pregabalin was commercially available.

[0096] In the embodiment of YET01, two doses are set, 20 mg / kg and 50 mg / kg respectively. The administration method is oral gavage, 10 ml / kg.

[0097] Dissolution of compounds:

[0098] YET01 was dissolved according to the following steps: (1) 20% of the injection volume of DMSO was completely dissolved; (2) 20% of the injection volume of polyethylene glycol 400 was added and mixed evenly; (3) 60% of the injection volume of sterile saline was diluted to the final volume. According to this operation sequence, each step was vortexed for 30 seconds, followed by ultrasonication for 1 minute, and the mixture was prepared and used on the same day.

[0099] Solvent control group: DMSO: polyethylene glycol 400: normal saline = 1:1:2

[0100] Positive control drug:

[0101] Pregabalin: The solvent is normal saline, and the administration method is intraperitoneal injection, 10ml / kg

[0102] Data analysis

[0103] All data analysis was performed using SPSS 22 (Formac) data processing software. Statistical comparisons were performed using one-way analysis of variance or two-way analysis of variance, and the results were verified by LSD multiple comparisons. Data are expressed as Mean ± SEM. When p < 0.05, one asterisk is marked; when p < 0.01, two asterisks are marked.

[0104] Experimental animals

[0105] 8-10 week old Balb / c or C57BL / 6J male mice, inbred strain (Inbred Mice), are internationally used experimental mice and were purchased from Shanghai Lingchang Experimental Animal Co., Ltd.

[0106] Example 2.1 Thermal stimulation tail flick experiment in mice (tail immersion)

[0107] In the mouse tail-flicking experiment, the mouse's tail is immersed in 52°C water. The mouse lifts its tail due to the pain caused by the harmful thermal stimulation. The mouse's pain tolerance is determined by timing the latency of tail-lifting. Figure 4 As shown, the results of this experiment indicate that YET01 can effectively relieve the pain caused by noxious thermal stimulation in mice.

[0108] Example 2.2 Cold stimulation tail-flick experiment in mice

[0109] In the mouse tail-flicking experiment, the mouse's tail is immersed in 5°C water. The pain caused by the harmful cold stimulation will also cause the mouse to lift its tail. The mouse's pain tolerance can be determined by timing the latency of tail lifting. Figure 5 As shown, the results of this experiment indicate that YET01 can effectively increase the tail-flick latency time of mice induced by noxious cold stimulation.

[0110] Example 2.3 Mouse chronic inflammatory pain model

[0111] Injecting 20 μl of Complete Freund's Adjuvant (CFA) into the soles of mice can produce redness, swelling, heat, and pain to simulate chronic inflammatory pain. 48-72 hours after modeling, Von Frey silk is used to measure the tolerance response of the model mice to triggered mechanical pain. Use a Von Frey silk kit (which can provide a stimulation force of 0.008g-300g), select a nylon silk of appropriate thickness according to actual conditions, and vertically stimulate the inflamed skin of the soles of the feet for 5 seconds. When mice are mechanically stimulated, they will reflexively retract their feet, and the intensity of the stimulation force used when lifting the paw is recorded. The value of 3 responses in 5 measurements is the pain threshold of the mouse triggered mechanical pain. Figure 6 As shown, oral administration of YET01 can increase the pain threshold 1-3 hours after administration, indicating that the drug can effectively alleviate the triggered mechanical pain response of CFA model mice.

[0112] Case study 2.4 Spontaneous pain assessment experiment in mice using a neuropathic pain model (Spared nerve injury, SNI)

[0113] SNI neuropathic pain mice showed spontaneous paw retraction and licking behaviors. The mice were recorded for 5 minutes in a quiet state using a camera, and the number of rapid paw lifting was counted as an indicator for evaluating spontaneous pain. Figure 7 As shown, YET01 or control solvent was administered by gavage. The results showed that 2 hours after administration of YET01, the count of spontaneous paw withdrawal of mice was significantly reduced, indicating that YET01 can significantly improve the spontaneous pain symptoms of neuropathic pain.

[0114] Example 2.5 Mouse neuropathic pain SNI model, cold plate test

[0115] The SNI model of neuralgia is characterized by a hyperalgesic response to cold painful stimuli. In this experiment, a cold plate was used to evaluate the tolerance response of the mouse neuralgia model to noxious cold stimuli. The cold plate is a borosilicate glass plate (6 mm thick). (1) The mice are acclimatized in a separate compartment with a size of 4”×4”×11”, and the adjacent parts are black and opaque; (2) Prepare cold stimuli: crush dry ice stored at -80°C into powder, cut off the tip of a 3mL BD brand syringe, put the powdered dry ice into the modified syringe and compact it, and trim the open tip to a smooth surface with a diameter of 1 cm; (3) The mice are tested when they are quiet but not asleep, and the hind feet of the mice are required to be in direct contact with the glass plate to avoid gaps; push the syringe The needle plug is placed so that the dry ice is higher than the end of the syringe and is lightly pressed against the bottom of the glass plate; (4) A stopwatch is used to record the latency of paw withdrawal, where the time interval between two consecutive tests is ≥7 minutes, and the interval between series of tests at two time points is ≥15 minutes; (5) Three consecutive measurements are used as the basic control value. In view of the time limit of the experimental stage, the hind paw is measured 1-2 times at each time point; (6) In order to reduce frostbite, the maximum latency of paw withdrawal is 30 seconds, that is, if there is no paw withdrawal reaction within 30 seconds, the time is recorded as the cutoff value. Figure 8 As shown in the figure, the results of this experiment show that oral administration of YET01 at 20 mg / kg and 50 mg / kg can significantly relieve the cold pain sensitivity symptoms of mice, which lasts for about 3 hours after administration. The drug efficacy is comparable to that of the positive drug pregabalin at 30 mg / kg, which is administered intraperitoneally.

[0116] In summary, YET01 can reduce pain responses in acute pain models, including noxious heat pain, noxious cold pain, chronic inflammatory pain, neuropathic pain and other related symptoms, including spontaneous pain and cold pain sensitivity models, showing that YET01 has significant anti-multiple pain effects. All documents mentioned in the present invention are cited as references in this application, just as each document is cited as a reference separately. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

[0117] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the attached claims.

Claims

1. Use of a compound as represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating pain, characterized in that: The structure of the compound is shown below: The pain is noxious heat pain, noxious cold pain, and chronic inflammatory pain.

2. The use according to claim 1, characterized in that The dosage form of the medicine is an oral dosage form or an injection, and the oral dosage form includes tablets, capsules, films, and granules.

3. The use according to claim 1, characterized in that The medicine also contains a pharmaceutically acceptable carrier.

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