Cognitive impairment models, uses, and the application of compound MT-1207 in the prevention and treatment of cognitive impairment

Abstract

This invention relates to the field of biomedical technology, specifically disclosing a cognitive impairment model, its uses, and the application of compound MT-1207 in the prevention and treatment of cognitive impairment. The structure of compound MT-1207 is shown in formula (I). This invention is the first to discover that 2K2C rats can serve as a cognitive impairment model and can be used for screening related drugs for cognitive impairment. Compound MT-1207 can improve cognitive function impairment in 2K2C rats, significantly improving the rats' movement distance, latency, number of platform crossings, and time spent in the target quadrant, and can improve the rats' learning and memory abilities.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically, to cognitive impairment models, uses, and the application of compound MT-1207 in the prevention and treatment of cognitive impairment. Background Technology

[0002] MT-1207, chemically named 3-(4-(4-(1h-benzotriazol-1-yl)butyl)piperazin-1-yl)benzisothiazole hydrochloride, is a novel antihypertensive drug that has entered clinical trials in China. A patent titled "New Uses of Multi-Target Compound X7" (2019109311600) discloses the use of MT-1207 for the treatment of hypertension and target organ damage caused by hypertension. Patents regarding the use of benzopentazoheterocyclic piperazine derivatives (2012104869676, 2016108284104) disclose the use of MT-1207 for the treatment of diseases or conditions related to persistent pathological vasoconstriction or vasospasm. The 2021 paper, "Pharmacological characterization of MT-1207, a novel multitarget antihypertensive agent," disclosed the pharmacological properties, efficacy, and related therapeutic targets of MT-1207, demonstrating that MT-1207 is a novel and promising single-molecule multitarget antihypertensive drug.

[0003] Cognition is the intelligent processing of knowledge acquisition by an organism, involving behaviors such as learning, memory, language, thinking, mental state, and emotion. Cognitive impairment refers to abnormalities in the higher-level intelligent processing of the brain related to learning, memory, and judgment, leading to learning and memory difficulties. Cognition is a normal brain function; any factor that causes abnormalities in brain function and structure can lead to cognitive impairment. As the world rapidly ages, cognitive impairment, Alzheimer's disease, and other diseases have become significant risk factors threatening human health in old age and reducing the quality of life for the elderly, imposing a heavy economic burden and psychological stress on society and families. Therefore, developing drugs to improve cognitive function has become a key research focus for medical professionals.

[0004] Renal hypertension is common in humans. Renal vascular hypertension models are created by vascular stenosis, occlusion, or organ damage, which impairs blood flow or increases resistance, leading to elevated blood pressure and kidney damage. The model is established by surgically separating the renal arteries under animal anesthesia to narrow them. Surgical methods include two-kidney clamping (2K1C), one-kidney clamping (1K1C), and two-kidney clamping (2K2C). Clamping one renal artery reduces blood flow by more than 50%, clamping one renal artery and then removing the contralateral kidney after a period of time, or simultaneously narrowing both renal arteries. In practice, various methods can be used to achieve renal artery stenosis, including ligation, suture embolization, and balloon angioplasty, and these methods are applied to different animals. Dogs were initially used as a model for renal hypertension, but it has since been applied to various animals such as rats, rabbits, dogs, pigs, cats, monkeys, and mice. Currently, the rat model is the most widely used. Using 2K2C animals (bilateral renal artery stenosis animals, i.e., animals whose bilateral renal arteries are narrowed through physical means, reducing renal blood flow by more than 50%) as animal models of hypertension for drug screening is an existing technology, and there are numerous related literature reports. For example, the 2005 paper "Establishment and Postoperative Management of a Bilateral Renal Artery-Clipped Renal Vascular Hypertension Model" publicly disclosed the use of bilateral renal artery-clamping to create a rat model of renal vascular hypertension.

[0005] However, in the existing technology, there are currently no reports on the use of 2K2C rats as an animal model of cognitive impairment for drug screening related to cognitive impairment, or on the application of MT-1207 in the prevention and treatment of cognitive impairment. Summary of the Invention

[0006] The purpose of this invention is to address the shortcomings of existing technologies by providing a new animal model of cognitive impairment, the application of this animal model in drug screening for cognitive impairment, and the application of the known compound MT-1207 in the prevention and treatment of cognitive impairment.

[0007] The first objective of this invention is to address the shortcomings of the prior art by providing the use of compound MT-1207 in the preparation of medicaments for the prevention and / or treatment of cognitive impairment, the structure of which is shown in formula (I).

[0008]

[0009] A second objective of this invention is to address the shortcomings of the prior art by providing the use of compound MT-1207 in the preparation of medicaments for improving learning and / or memory abilities.

[0010] In the above applications, preferably, the compound MT-1207 is the sole active ingredient.

[0011] In the above applications, preferably, the compound MT-1207 is used as an active ingredient in combination with other drugs for treating cognitive impairment.

[0012] In the above applications, preferably, the drug further includes one or more pharmaceutically acceptable excipients; the pharmaceutically acceptable excipients are excipients, fillers, or diluents.

[0013] More preferably, in the above applications, the drug comprises MT-1207 in a weight ratio of 0.01 to 99.99% and an excipient in a weight ratio of 0.01 to 99.99%.

[0014] More preferably, in the above applications, the drug comprises MT-1207 in a weight ratio of 0.01 to 99.99%, and other drugs for treating cognitive impairment in a weight ratio of 0.01 to 99.99%.

[0015] In the above applications, preferably, the drug is prepared into a clinically acceptable pharmaceutical formulation according to conventional pharmaceutical preparation methods.

[0016] In the above applications, preferably, the drug is in various dosage forms conventional in the art, more preferably in solid, semi-solid, or liquid form, and can be an aqueous solution, non-aqueous solution, or suspension, more preferably in powder, granule, tablet, capsule, pill, solution, suspension, or injection. The preferred route of administration is injection or oral administration. Preferred routes of injection include intravenous injection, intramuscular injection, intraperitoneal injection, intradermal injection, or subcutaneous injection.

[0017] The third objective of this invention is to address the shortcomings of the prior art by providing the application of 2K2C animals in the preparation of animal models of cognitive dysfunction.

[0018] In the above applications, preferably, the animal is a rat, rabbit, dog, pig, cat, monkey, or mouse.

[0019] The fourth objective of this invention is to address the shortcomings of the prior art by providing 2K2C animals as animal models of cognitive dysfunction for the screening of related drugs.

[0020] In the above applications, preferably, the animal is a rat, rabbit, dog, pig, cat, monkey, or mouse.

[0021] The advantages of this invention are:

[0022] 1. This study is the first to discover that 2K2C rats can serve as a model of cognitive impairment and can be used for screening related drugs for cognitive impairment.

[0023] 2. For the first time, it was discovered that compound MT-1207 can improve cognitive function impairment in 2K2C rats, significantly improving the rats' movement distance, latency, number of platform crossings, and time spent in the target quadrant, and can improve the rats' learning and memory abilities. Attached Figure Description

[0024] Figure 1 Morris Water Maze Experiment Video Acquisition and Analysis System. A. Main body of the water maze; B. Computer software system; C. Lighting conditions; D. Quadrant division. The small red circle in the 3rd quadrant indicates the platform location, and the scale represents the diameter of the water maze, which is 1.6 meters.

[0025] Figure 2 Effects of MT-1207 on the water maze test in rats. A. Day 1–5, total swimming distance; B. Day 1–5, latency to reach the platform; C. Day 6, number of times the platform was crossed; D. Day 6, time spent in the target quadrant; E. Day 6, swimming speed; F. Day 6, rat movement trajectory diagram; n = 10–16. * P<0.05 ** P<0.01 *** P<0.001VS Model. Detailed Implementation

[0026] The present invention will be further described below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the description of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0027] In this invention, the compound MT-1207 can be obtained commercially or prepared using a disclosed method. The structure of compound MT-1207 is shown in formula (I).

[0028]

[0029] This invention first prepared 2K2C rats and used them as an animal model of cognitive impairment for drug screening related to cognitive impairment. Animal experiments showed that in the navigation experiment, after swimming training, the total swimming distance and latency to reach the platform in the Sham group gradually decreased, while the total swimming distance and latency of the 2K2C rats were significantly higher than those in the Sham group, indicating that the 2K2C rats had learning and memory impairment. In the spatial exploration experiment, the number of times the 2K2C rats crossed the platform area and the time spent in the target quadrant were significantly lower than those in the normal group, indicating that the 2K2C rats had memory decline. The swimming speed of both the normal Sham group rats and the 2K2C rats was basically the same, indicating that there was no motor impairment when using the 2K2C rats as a model animal. The above experimental results show that the 2K2C rat can be used as an animal model of cognitive impairment and can be used for drug screening related to cognitive impairment.

[0030] Animal experiments with compound MT-1207 showed that in the positioning and navigation experiment, the MT-1207-treated group significantly reduced the distance traveled and latency compared to the model group, indicating that MT-1207 has an effect on improving learning and memory function in the 2K2C rat model. In the spatial exploration experiment, the MT-1207-treated group traversed the platform area more times than the model group, and the MT-1207 10mg / kg and 20mg / kg groups spent significantly more time in the target quadrant than the model group. The MT-1207-treated group rats exhibited higher memory capacity, repeatedly exploring the target area and locating the underwater platform. These experimental results demonstrate that compound MT-1207 can improve cognitive impairment and enhance learning and memory abilities.

[0031] The 2K2C animals of this invention are commonly used hypertension model animals. The term "animal" refers to mammals, preferably rats, rabbits, dogs, pigs, cats, monkeys, and mice, with rats being more preferred. 2K2C animals can be obtained commercially, prepared using methods reported in existing literature, or prepared using the methods described in the embodiments of this invention. The methods for preparing 2K2C animals (bilateral renal artery stenosis animals) are common knowledge. For example, bilateral renal arteries in model animals can be narrowed physically, reducing blood flow by more than 50%. Physical methods include ligation, suture occlusion, U-shaped silver clips, and balloon methods to narrow the bilateral renal arteries. A specific embodiment of this invention details the preparation of 2K2C rats by narrowing the bilateral renal arteries of rats using U-shaped silver clips. Those skilled in the art can also prepare more 2K2C animals using different methods based on the modeling mechanism of bilateral renal artery stenosis animal models, and use them as animal models of cognitive impairment according to the principles of this invention for screening related drugs for cognitive impairment. These applications are all within the scope of protection of this invention.

[0032] The compound MT-1207 of this invention can be formulated into suitable pharmaceutical compositions for oral administration, parenteral administration, inhalation spray administration, rectal administration, intranasal administration, sublingual administration, buccal administration, transdermal administration, or implantation administration. Parenteral administration includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. The compound MT-1207 of this invention is preferably administered orally, sublingually, intravenously, intramuscularly, or subcutaneously. The pharmaceutical composition may contain one or more conventional pharmaceutically acceptable carriers, excipients, or media, such as: diluents, excipients like water; binders such as cellulose derivatives, gelatin, polyvinylpyrrolidone, etc.; fillers such as starch; disintegrants such as calcium carbonate, sodium bicarbonate; lubricants such as calcium stearate or magnesium stearate; and other adjuvants such as flavoring agents and sweeteners.

[0033] Pharmaceutical compositions containing the compound MT-1207 of the present invention can be in the form of sterile injectable formulations, for example, as sterile aqueous or oily suspensions. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. Sterile injectable formulations can also be sterile injectable solutions or suspensions in non-toxic diluents or solvents suitable for parenteral administration, for example, as solutions in 1,3-butanediol. Suitable media and solvents include mannitol, water, and isotonic sodium chloride solutions. Furthermore, sterile non-volatile oils are conventionally used as solvents or suspension media. Any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, can be used in injectable formulations, as can naturally pharmaceutically acceptable oils, such as olive oil or castor oil, especially their polyoxyethyleneized forms. The oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants or similar alcohols.

[0034] Pharmaceutical compositions containing the compound MT-1207 of this invention can be administered orally in any orally available dosage form, including but not limited to capsules, tablets, powders, granules, aqueous suspensions, and solutions. These dosage forms are prepared according to techniques well known in the field of pharmaceutical formulation. For tablets used orally, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in capsule form, available diluents include lactose and dried corn starch. When an aqueous suspension is administered orally, the active ingredient may be combined with an emulsifier and a suspending agent. If desired, certain sweeteners and / or flavoring agents and / or coloring agents may be added.

[0035] Pharmaceutical compositions containing the compound MT-1207 of the present invention can be administered via nasal aerosol or inhalation. Such compositions can be prepared according to techniques well known in the field of pharmaceutical formulation and can be prepared as solutions in saline using benzyl alcohol or other suitable preservatives, bioavailability enhancers, fluorocarbons, and / or other solubilizers or dispersants known in the art.

[0036] Pharmaceutical compositions containing the compound MT-1207 of the present invention can also be administered in the form of suppositories for rectal administration. The compositions can be prepared by mixing the compound of the present invention with a suitable, non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and thus dissolves in the rectum to release the active ingredient. Such substances include, but are not limited to, cocoa butter, beeswax, and polyethylene glycol.

[0037] The pharmaceutical composition containing the compound MT-1207 of the present invention can be prepared by conventional methods in the medical field, wherein the weight ratio of the active ingredient is 0.01 to 99.99%, which depends on the condition to be treated or prevented and the characteristics of the subject to whom the compound is administered. The dosage regimen for the given compound MT-1207 can be easily determined by those skilled in the art using the content disclosed herein.

[0038] The compound MT-1207 of the present invention can be used in combination with one or more other active pharmaceutical components. The combination drug can be in the form of a single composition containing the compound MT-1207 of the present invention and one or more other active pharmaceutical components, or in the form of a combination of two or more separate compositions.

[0039] Examples

[0040] 1. Experimental purpose

[0041] For the first time, 2K2C rats were used as a cognitive dysfunction model, and for the first time, the effect of MT-1207 (drug code) on improving cognitive function was investigated.

[0042] 2. Experimental materials

[0043] 2.1. Test samples

[0044] MT-1207 (abbreviation: MT): white or quasi-white crystalline powder, production batch number: 20181201, stored sealed in a 4°C refrigerator.

[0045] 2.2. Experimental reagents

[0046] Pentobarbital sodium: produced by Merck, Germany, imported and repackaged.

[0047] 2.3. Experimental animals

[0048] SD rats, SPF grade, body weight 120 - 150 g, male, number 80, sold by: Experimental Animal Business Department of Shanghai Institute of Family Planning, production license number: SCXK (Shanghai) 2018 - 0006.

[0049] 2.4. Experimental instruments

[0050] U-shaped silver clip: produced by Shanghai Alcott Biotech Co., Ltd.

[0051] Morris water maze analysis system: model XR-XM101, produced by Shanghai Xinruan Technology Co., Ltd.

[0052] 3. Experimental methods

[0053] 3.1. Grouping, dose setting and administration method

[0054] Sham surgery group: fed regular feed;

[0055] 2K2C Model Group: Feeded with regular feed;

[0056] 2K2C model MT-1207 treatment group 1 (MT 10mg / kg): The dose was 10mg / kg, and the administration method was feeding medicated feed;

[0057] 2K2C model MT-1207 treatment group 2 (MT 20mg / kg): The dose was 20mg / kg, and the administration method was feeding medicated feed.

[0058] 3.2.2 K2C Model Preparation

[0059] Rats were fasted for 12 hours prior to surgery but allowed free access to water. Model preparation began the following day. Rats were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (40 mg / kg) at a dose of 2 ml / kg. The rats were then fixed in a supine position on a rat table. The chest and abdominal area was shaved, and the skin surface was disinfected with povidone-iodine and 75% ethanol. An incision was made 1.5 cm below the xiphoid process, extending approximately 1.5 cm distally along the midline of the abdomen. The muscle layer was cut along the linea alba, and the incision was opened with a retractor to expose the surgical field. The retractor was gently lifted upwards, and a moistened sterile cotton swab was used to push aside the organs and tissues above and around the right kidney. A cotton ball was then used to push aside the tissues above and below the kidney to fully expose the renal vessels. The kidney was turned inside out with a cotton swab, and the surrounding adipose tissue was gently separated with forceps to expose the renal artery. The renal artery was carefully separated, and a 0.2 mm inner diameter U-shaped silver clip was placed on it. All tissues were then repositioned. The contralateral renal artery was separated and clipped using the same method. After repositioning all tissues, 40,000 to 80,000 units of penicillin sodium were dripped in, and then the muscles and skin were sutured layer by layer. The animals were kept warm on an electric blanket until they regained consciousness, after which they were returned to their cages for rearing. The procedure for the sham-operated group of rats was similar to that for the 2K2C model, but only the bilateral renal arteries were freed, and U-shaped silver clips were not applied.

[0060] 3.3. Water Maze Experiment

[0061] Two weeks after surgery, 2K2C model rats were randomly divided into three groups: a model control group and two drug treatment groups. A sham-operated normal control group was also included. Drug treatment lasted for 8 weeks, followed by the water maze test.

[0062] The water maze experiment consists of two parts: a positioning and navigation experiment and a space exploration experiment. This experiment is conducted daily from 19:00 to 23:00. Before the experiment begins, four markers of different shapes are suspended at appropriate positions on the water maze's suspension frame. Water is then poured into the pool until the water level is approximately 1 cm above the platform. The heating system is turned on, and the water temperature is set to 22℃. Four table lamps are placed at equal intervals on the ground surrounding the water maze, positioned approximately 50 cm outside the markers, with the light directed directly upwards to create uniform diffused light within the water maze. During the test, the room's main lighting system must be turned off, and the table lamps must provide illumination to maintain a relatively dim environment inside the water maze. Figure 1 AC).

[0063] On the computer, adjust the initial parameters for the water maze experiment. First, draw a circular area around the edge of the water surface, setting the diameter of the circle to 1.6m. Then, divide the circle into four equal regions, naming the upper right corner as the first quadrant, and naming the four quadrants clockwise. Next, trace the outline of the platform in the third quadrant, naming it the platform area. Figure 1 D) Set the video recording duration to 60 seconds, and the trigger condition for stopping recording is entering the platform area and staying there for 3 seconds. After setting, use black dye to turn the water in the basin black, which will create a sharp contrast with the white color of the rat, helping the video analysis system to accurately capture the rat's timely location.

[0064] Orientation and Navigation Experiment: This experiment lasted for 5 days. Each day, rats were gently placed into the water from each of the four quadrants, with their heads facing the pool wall. The rats were allowed to explore freely, and a video capture and analysis system recorded their swimming trajectories in real time. If a rat did not find the hidden platform within 60 seconds, it was guided to the platform and allowed to stay for 15 seconds to help it memorize the location. After the experiment, the rats were removed and placed in a specially designed cage, and their fur was dried with a warm air blower. After all rats had been trained from the same entry point, the training was repeated at the entry point in the next quadrant. This experiment primarily examines the latency and total distance traveled by the rats to find the platform, used to assess their spatial perception (orientation) and memory abilities.

[0065] Spatial exploration experiment: On day 6, the underwater platform was removed, and the experiment began 24 hours after the last training session. Rats were gently placed into the water facing the pool wall from the quadrant opposite the platform (quadrant 1 in this experiment), and their movement trajectory was recorded over 60 seconds. This experiment primarily examines the number of times the rats crossed the platform and the percentage of time spent in the target quadrant, used to assess the rats' memory ability.

[0066] 4. Experimental Results

[0067] In the navigation experiment, after 5 days of swimming training, the rats became increasingly faster at finding the platform hidden underwater. Regardless of whether it was the Sham group or any other group, the total swimming distance and the latency to reach the platform gradually shortened. Figure 2 As shown in Figures A and B, on days 4 and 5 of training, the movement distance and latency of the Model group rats were significantly higher than those of the Sham group, indicating that the Model group rats had learning and memory dysfunction. Compared with the Model group, the two MT-1207 administration groups significantly reduced movement distance and latency, indicating that MT-1207 has an effect on improving learning and memory function in the 2K2C rat model.

[0068] On day 6, a space exploration experiment was conducted. The underwater platform was removed, and the process of the rats searching for the platform using their own memory within 1 minute was observed. Figure 2 F). The tests revealed that, compared to the normal group, the model group had a reduced number of times it traversed the platform region and a decreased time spent in the target quadrant, suggesting a decline in memory function in the 2K2C model. The number of times rats in both MT-1207 treatment groups traversed the platform region was higher than that in the model group, and the difference was significant between the MT-1207 20mg / kg group and the model group. Regarding the exploration time in the target quadrant, the MT-1207 10mg / kg and 20mg / kg groups had significantly longer time spent in the target quadrant than the model group. Figure 2 (C, D). These two results suggest that the rats in the MT-1207 treatment group have high memory ability and are able to repeatedly explore the target area and find the underwater platform.

[0069] Simultaneously, we also tested the swimming speed of the rats in each group. This test aimed to determine if there were any defects in the animals' motor abilities and to rule out interference from motor impairments in learning and memory tests. The tests revealed that the swimming speeds of the rats in each group were basically the same, indicating no motor impairments. Figure 2 E).

[0070] In conclusion, 2K2C rats can serve as a model of cognitive impairment. MT-1207 can improve cognitive impairment in 2K2C rats, significantly improving the rats' movement distance, latency, number of platform crossings, and time spent in the target quadrant, and can improve the rats' learning and memory abilities.

[0071] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several improvements and additions without departing from the method of the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention.

Claims

1. Use of a compound MT-1207, whose structure is shown in formula (I), for the preparation of a medicament for preventing and / or treating cognitive dysfunction, said cognitive function being learning and memory function, 2. Use according to claim 1, characterized in that, said compound MT-1207 as the only active ingredient.

3. Use according to claim 1, characterized in that, said compound MT-1207 as the active ingredient together with other drugs for treating cognitive dysfunction.

4. Use according to claim 1, characterized in that, said medicament further comprising one or more pharmaceutically acceptable auxiliary ingredients.

5. The use according to claim 1, characterized in that, said medicament prepared into a clinically acceptable pharmaceutical preparation according to the conventional method for preparing medicaments.

6. Use according to claim 5, characterized in that, said pharmaceutical preparation being granules, capsules, tablets, suspensions, emulsions, solutions or injections, and said medicament can be applied to the individual in need of treatment through different dosage forms by oral, intravenous, intramuscular or subcutaneous injection.

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