Use of compound P57 or its analogs for lowering body temperature and protecting nerves

Abstract

The present invention provides the use of compound P57 or its analogs for hypothermia and neuroprotection. Specifically, the present invention provides the use of a compound represented by Formula I below for the preparation of a pharmaceutical composition for lowering the body temperature of a subject. The compound can be used for neuroprotection against hypoxic-ischemic diseases. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the neuroprotective effects of P57 and its analogs in lowering body temperature, treating hypoxic-ischemic diseases, and their application in fever. Background Art

[0002] Natural products are derived from animals, plants, microorganisms, or chemical components of these organisms. They primarily include flavonoids, anthraquinones, saponins, alkaloids, terpenes, peptides, polysaccharides, and proteins. Natural products, especially medicinal plants, remain an important source of new chemical entities (NCEs).

[0003] Physical cooling has been used as an emergency treatment for many acute hemorrhagic conditions, but rapid and specific hypothermia induced by small molecules has not yet been clinically applied. Hibernation is a stress-resistant mechanism used by organisms to cope with extreme cold, and lowering body temperature in hibernating animals has a protective effect. In 1987, Busto and colleagues discovered that lowering brain temperature by just a few degrees could reduce neuronal death. This discovery sparked interest in the neuroprotective effects of hypothermia, and subsequent experiments further demonstrated its profound neuroprotective effects. Therapeutic hypothermia has become one of the most reliable neuroprotective approaches (blocking cell apoptosis and necrosis) for several brain diseases and injuries, such as stroke, traumatic brain injury, and global cerebral ischemia and hypoxic-ischemic encephalopathy after cardiac arrest. Stroke is a leading cause of death and disability, with approximately 87% of strokes being ischemic. Despite its high prevalence, the limited window of opportunity for intervention limits the number of patients who can receive it. Therapeutic hypothermia can provide potent neuroprotective effects, making it a potentially attractive treatment option for ischemic stroke.

[0004] Therefore, there is still a need in the art to develop new hypothermic neuroprotective agents for the neuroprotective effect in treating hypoxic-ischemic diseases. Summary of the Invention

[0005] The purpose of the present invention is to provide a compound P57 and its analogs for preparing a pharmaceutical composition for lowering body temperature or protecting neurons.

[0006] In a first aspect of the present invention, a use of compound P57 or an analog thereof is provided, characterized in that it is used to prepare a pharmaceutical composition for lowering the body temperature of a subject, and the compound P57 or an analog thereof has a structure as shown in Formula I below:

[0007]

[0008] wherein R1 is selected from the group consisting of hydrogen, C2-C6 alkanoyl, unsubstituted or para-substituted benzoyl, tri(C3-C9 alkyl)silyl, tri(C9-C 16 aryl)silyl, allyl, or substituted or unsubstituted benzyl;

[0009] R2 is selected from hydrogen, hydroxyl, halogen; wherein the hydroxyl may have a protecting group, and the protecting group is selected from the following group: C2-C6 alkanoyl, unsubstituted or para-substituted benzoyl, tri(C3-C9 alkyl)silyl, tri(C9-C 16 aryl)silyl, allyl, or substituted or unsubstituted benzyl;

[0010] Wherein, the substituent of the benzoyl group is selected from the following group: methoxy, nitro, azido, halogen; the substituent on the benzyl group is selected from the following group: naphthylene, p-methoxyphenyl, p-methylphenyl, p-nitrobenzyl, and para-halogen-substituted benzyl.

[0011] In another preferred embodiment, the R1 is selected from the following group: acetyl, unsubstituted benzoyl, or benzoyl substituted with p-methoxy.

[0012] In another preferred embodiment, the compound is selected from the following group:

[0013]

[0014]

[0015]

[0016] In another preferred embodiment, the compound is P57:

[0017]

[0018] In another preferred embodiment, the compound is 6-70 or 2,74-157-α:

[0019]

[0020] In another preferred embodiment, the pharmaceutical composition is also used to treat or prevent hypoxic-ischemic diseases.

[0021] In another preferred embodiment, the pharmaceutical composition is used for neuronal protection in the treatment of hypoxic-ischemic diseases.

[0022] In another preferred embodiment, the hypoxic-ischemic disease is selected from the group consisting of stroke, traumatic brain injury, global cerebral ischemia after cardiac arrest, and hypoxic-ischemic encephalopathy.

[0023] In another preferred embodiment, the pharmaceutical composition is also used to treat or improve fever symptoms of the subject.

[0024] In another preferred embodiment, the pharmaceutical composition is an injection.

[0025] 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 described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Temperature changes over time in mice and rats after intraperitoneal injection of P57. (A) 6-8 week old male C57BL / 6J mice were intraperitoneally injected with different doses of P57: 12.5 mg / kg and 25 mg / kg, with the vehicle group serving as the blank control, n = 5; (B) 6-8 week old male Wistar rats were intraperitoneally injected with different doses of P57: 8.75 mg / kg and 17.5 mg / kg, with the vehicle group serving as the blank control, n = 4.

[0027] Figure 2 Effects of hypothalamic injection of P57 on body temperature in C57BL / 6J mice. (A) Schematic diagram of hypothalamic injection in C57BL / 6J mice. (B) Curves showing changes in body temperature over time in C57BL / 6J mice after hypothalamic injection of P57. Male C57BL / 6J mice were implanted with a double-barreled microinjection system in the hypothalamus. Two weeks later, different doses of P57 (15 μg and 30 μg) were injected directly into the hypothalamus. Anal body temperature was measured every 30 minutes. The vehicle group served as a blank control. N = 3.

[0028] Figure 3 P57 has a neuroprotective effect in the MCAO model. (A) Body temperature changes after intraperitoneal injection of P57 in the MCAO model. (B) 24 hours after intraperitoneal injection of P57 in the MCAO model, brain tissue samples were collected for TTC staining and infarct volume was calculated.

[0029] Figure 4 The role of P57 in a myocardial ischemia model. (A) Vevo 2100 microultrasound system was used to assess the effects of P57 treatment on mouse cardiac function. Ejection fraction (EF) and fractional shortening (ES) were measured. (B) Masson staining was used to assess the protective effect of P57 against myocardial ischemia.

[0030] Figure 5 P57 has the effect of lowering animal body temperature in the PGE2-induced fever model.

[0031] Figure 6Figure 3. Temperature changes over time in mice after intraperitoneal injection of P57 analogs. (A) 6-8 week old male C57BL / 6J mice injected intraperitoneally with 25 mg / kg 6-70, n=4; (B) 6-8 week old male C57BL / 6J mice injected intraperitoneally with 25 mg / kg 2,74-157-α, n=5. DETAILED DESCRIPTION

[0032] After extensive and in-depth research, the inventors unexpectedly discovered that P57, a drug compound known in the art for weight loss, can significantly lower body temperature in animals after administration. This discovery, combined with its potential as a hypothermic neuroprotectant, can provide neuroprotection in the treatment of hypoxic-ischemic diseases. Based on this discovery, the inventors completed the present invention.

[0033] P57 compounds and their neuroprotective effects

[0034] As used herein, "P57" refers to a compound having the following structure:

[0035]

[0036] P57 is a steroidal glycoside compound isolated from the African butterfly cactus. In recent years, P57 has attracted considerable attention as a health supplement with weight-loss effects. However, due to its unclear mechanism of action, it has not yet been applied in the medical and health care industry. In their research on the hypothalamic regulatory function of P57, the applicant discovered that this type of compound can rapidly induce a decrease in rodent body temperature. This phenomenon suggests that this type of compound may act on specific molecular targets in vivo, exerting central regulatory activity.

[0037] Pharmaceutical compositions and methods of administration

[0038] Since compound P57 has excellent activity in lowering the body temperature of the treated subject, the compound of the present invention and its various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compound of the present invention as the main active ingredient can be used to treat diseases caused by hyperthermia, such as for the treatment of fever, or for neuroprotection in hypoxic-ischemic diseases.

[0039] The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention and a pharmaceutically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-200 mg per dose. Preferably, "one dose" is one capsule or tablet.

[0040] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0041] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration, or subcutaneous administration).

[0042] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0043] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0044] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0045] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0046] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0047] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0048] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (eg, other neuroprotective drugs).

[0049] When administered in combination, the pharmaceutical composition may also include one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds. One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compounds may be administered simultaneously, separately, or sequentially with the compound of the present invention.

[0050] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0051] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0052] Example 1: P57 induces hypothermia in mice and rats

[0053] 1.1 Materials

[0054] 1.1.1 C57BL / 6J 6-8 week old male mice

[0055] 1.1.2 Wistar male rats aged 6-8 weeks

[0056] 1.1.3 Dimethyl sulfoxide (DMSO)

[0057] 1.1.4 Castor Oil

[0058] 1.1.5 Phosphate Buffered Saline (PBS)

[0059] 1.1.6 75% ethanol solution

[0060] 1.1.7 Lincomycin Lidocaine Gel

[0061] 1.1.8 Povidone-iodine solution

[0062] 1.1.9 Mouse double-tube microdosage system

[0063] 1.1.10 1mL disposable sterile syringe

[0064] 1.1.11 5μL microsyringe

[0065] 1.1.12 1.5mL EP tube

[0066] 1.1.13 15mL centrifuge tube

[0067] 1.2 Equipment

[0068] 1.2.1 Mouse rectal temperature detector

[0069] 1.2.2 Desktop digital stereotaxic instrument

[0070] 1.2.3 Mouse Adapter

[0071] 1.2.4 Mini round head drill

[0072] 1.3 Solution

[0073] 1.3.1 P57 was dissolved in 1% DMSO + 9% castor oil + 90% PBS solution.

[0074] 1.3.2 Preparation of 5% water and chloral solution: Dissolve 2.5 g water and chloral in 50 mL 1× PBS solution.

[0075] 1.4 Methods

[0076] 1.4.1 Hypothalamic implantation of a double-tube microdrug delivery system

[0077] The toes of 6-8 week C57BL / 6J mice were clipped and numbered, weighed, and anesthetized by intraperitoneal injection of 5% water and chloral (0.1mL / 10g). After the mice were in anesthesia, the hair in the surgical area of ​​the mouse head was shaved with a pet razor, and the scalp was cleaned with povidone iodine solution. The mice were fixed on a stereotaxic instrument, and the skin was incised along the midline of the head with a scalpel. The incision size should be appropriate. The skull surface was wiped clean with PBS dipped in cotton swabs and blown dry with ear wash bulbs to make the sutures clearly visible. The brain stereotaxic instrument was adjusted to make the skull level. Holes were made on the skull with a fine drill bit of a micro electric grinder around the target brain area and small stainless steel screws were implanted. Based on the hypothalamic region (M / L = ±0.40, A / P = -0.16, D / V = -5.15) as described in references and mouse brain atlases, a small hole was made in the skull using a fine drill bit from a micro-electric grinder, taking care not to damage the meninges or brain tissue. The microinjection system was then slowly implanted. Zinc sulfate cement was used to secure the system to the skull with a small stainless steel screw to enhance fixation. Mice were housed in spacious cages and, after 2 weeks, used for drug intervention experiments.

[0078] 1.4.2 P57 Processing

[0079] Intraperitoneal injection: According to the experimental design, mice / rats that need to be treated with drugs are randomly divided into: control group (89% PBS+10% castor oil+1% DMSO) and groups treated with different concentrations of P57.

[0080] Hypothalamic injection: Mice that had been implanted with a microdrug delivery system 2 weeks prior and were in good health were randomly divided into groups according to the experimental design. The hypothalamic injection dose was controlled at 2 μL.

[0081] 1.4.3 Body temperature measurement

[0082] For the first 3-4 days of temperature measurement, handle the mice several times daily at the same time to reduce stress from handling. Before drug intervention, measure the mice's temperature every 30 minutes, and measure basal body temperature for 2-3 hours. After drug intervention, measure the mice's temperature every 30 minutes, and monitor changes in body temperature for 8 hours.

[0083] 1.5 Results: P57 induces hypothermia in mice and rats

[0084] C57BL / 6J 6-8 week old male adult mice were intraperitoneally injected with different concentrations of P57 (12.5mg / kg, 25mg / kg) and solvent control, and the changes in the anal body temperature of the mice were monitored every 30 minutes. The results showed that the body temperature of the mice in the compound treatment group was significantly lower than that in the control group. About 1 hour after the injection of P57, the body temperature dropped to the lowest point (the body temperature of the 25mg / kg group dropped to about 32.0℃), and the degree of cooling and the duration of maintenance were positively correlated with the dose of the compound, while the body temperature of the mice in the control group was basically constant at about 37℃ (such as Figure 1 As shown in Figure A). Considering that mice have a larger specific surface area and their body temperature is more easily affected by external factors, we also used Wistar male adult rats to conduct the same verification. After intraperitoneal injection of different doses of P57 (8.75 mg / kg, 17.5 mg / kg) into Wistar rats, their body temperature was also significantly reduced (as shown in Figure A). Figure 1 (shown in B).

[0085] A double-tube microinjection system was implanted in the hypothalamus of adult C57BL / 6J male mice (x=±0.4, y=-0.16, z=-5.15). Two weeks after implantation, low doses of P57 (15 μg, 30 μg) were injected directly into the hypothalamus. Figure 2 The results showed that low-dose P57 induced a faster decrease in body temperature in mice (30 μg group, body temperature dropped to around 29°C 30 minutes after injection), and maintained the low temperature for up to 4-5 hours ( Figure 2 These data further demonstrate that P57 can induce hypothermia and suggest that P57 induces hypothermia by acting on the hypothalamus.

[0086] Example 2 Neuroprotective Effects of P57 in the Treatment of Hypoxic-Ischemic Diseases

[0087] 2.1 Materials

[0088] 2.1.1 2,3,5-Triphenyltetrazolium chloride

[0089] 2.1.2 Nylon monofilament suture

[0090] 2.2 Equipment

[0091] 2.2.1 Ventilator

[0092] 2.2.2 Vevo 2100 Micro-Ultrasound System

[0093] 2.3 Solution

[0094] 2.3.1 Preparation of 1% 2,3,5-triphenyltetrazolium chloride (TTC) solution: Weigh 1 g of TTC and dissolve it in 100 mL of 1× PBS solution.

[0095] 2.3.2 Preparation of 4% paraformaldehyde solution: Dissolve 4 g paraformaldehyde in 1× PBS, adjust pH to 7.4, dilute to 100 mL, and filter. Protect from light and store at 4°C until use.

[0096] 2.4 Methods

[0097] 2.4.1 Establishment of the rat middle cerebral artery occlusion (MCAO) model

[0098] Wistar rats were randomly divided into a sham-operated blank control group, a sham-operated drug-intervention group, a MCAO blank control group, and a MCAO drug-intervention group. Rats were weighed and recorded for body weight, and anesthetized with an intraperitoneal injection of 10% water and chloral. The neck skin was disinfected, and the left common carotid artery was isolated, taking care to avoid touching the vagus nerve within the carotid sheath. In the sham-operated group, only the blood vessels were isolated without inserting nylon suture. In the MCAO group, a punctate incision was made on the ventral wall of the common carotid artery using a 25g needle. A nylon monofilament suture (5 cm long, 0.43 mm diameter) with a silicone rubber tip was inserted 20 mm into the middle cerebral artery. The skin was sutured, and after 120 minutes of occlusion, the suture was removed, and perfusion of the middle cerebral artery was unobstructed. Simultaneously, drug intervention was administered intraperitoneally.

[0099] 2.4.2 TTC staining of rat cerebral infarction area

[0100] Twenty-four hours after intervention on P57, rats in the sham and MCAO groups were anesthetized with water and chloral, decapitated, and brains removed. Brains were quickly frozen at -20°C for 20 minutes and cut into 2-mm-thick coronal sections. The sections were immediately stained in 1% TTC solution at room temperature in the dark for 45 minutes. After washing with PBS, the sections were fixed with 4% paraformaldehyde and incubated at 4°C for 24 hours. Red areas indicate normal brain tissue, and white areas indicate brain tissue with infarcted lesions. Image J software was used to calculate the infarct area of ​​each sample.

[0101] 2.4.3 Mouse myocardial ischemia model

[0102] Mice were anesthetized with 5% isoflurane in an airtight chamber and intubated using a 100% O2 volume-controlled ventilator supplemented with 2.5% isoflurane. After thoracotomy and pericardectomy, the left anterior descending coronary artery (LAD) was found along the anterior wall of the left ventricle. The LAD artery was ligated with 7-0 polypropylene suture near its origin and the left main coronary artery. LAD artery occlusion immediately caused the anterior wall of the left ventricle to turn white, indicating myocardial ischemia. Subsequently, the chest skin was sutured with 6-0 sutures. The trachea was extubated and the mice were placed on a 37°C heating pad until they woke up. The ejection fraction of the mice was measured by echocardiography on the third, seventh, and fourteenth days, and then samples were taken and sections were stained with Masson.

[0103] 2.4.4 Mouse echocardiography

[0104] Cardiac function was assessed using a Vevo 2100 microultrasound system on days 3, 7, and 14 after myocardial ischemia in mice. Left ventricular ejection fraction (LVEF), left ventricular internal shortening (LVES), left ventricular internal diameter at diastole (LVIDd), and left ventricular internal diameter at systole (LCIDs) were recorded and calculated.

[0105] 2.5 Results

[0106] 2.5.1 P57 has a neuroprotective effect in the MCAO model

[0107] First, we established a rat model of middle cerebral artery occlusion (MCAO) to simulate stroke. P57 was injected intraperitoneally during 2 hours of ischemia and reperfusion in rats. The control group was injected intraperitoneally with solvent and the changes in body temperature of the rats were monitored ( Figure 3 (A) We found that P57 still had significant hypothermia-inducing activity in this model. After 24 hours of reperfusion, the rats were anesthetized with isoflurane and the brains were removed for 2,3,5-triphenyl tetrazolium chloride (TTC) staining. We observed that the volume of ischemic cerebral infarction in the brains of rats in the P57-treated group was significantly reduced compared with the control group (p<0.01, n=7). This result indicates that P57 can significantly reduce brain neuronal death and has a neuroprotective effect in the MCAO model ( Figure 3 Middle B).

[0108] 2.5.2 P57 has a protective effect in myocardial ischemia models

[0109] We established a myocardial ischemia model in mice by ligating the LAD coronary artery and treated them with P57 intraperitoneally for 7 days. The control group was injected with solvent. The cardiac function of the mice was evaluated using the Vevo 2100 micro-ultrasound system on the third, seventh, and fourteenth days. Figure 4 As shown in A, on the third and seventh days, the left ventricular ejection fraction (LVEF) and left ventricular wall shortening fraction (LVES) of the P57 treatment group were greater than those of the control group. Figure 4 Middle B) shows that the infarct size in the P57-treated group was smaller than that in the control group. These results indicate that P57 has a certain protective effect on the myocardial ischemia model.

[0110] Example 3 Effect of P57 in treating fever

[0111] 3.1 Materials

[0112] 3.1.1 Prostaglandin E2 (PGE2)

[0113] 3.2 Solution

[0114] 3.1.1 PGE2 solution: Dissolve PGE2 in saline to 2 M.

[0115] 3.3 Methods:

[0116] Fever Model

[0117] Mice in good health were randomly divided into a blank control group, a P57 group, a PGE2 group, and a P57+PGE2 group. Mice were weighed and their body weights were recorded. Body temperature was recorded in metabolic cages starting 1 hour before drug treatment. P57 (25 mg / kg) was injected intraperitoneally in the P57 and P57+PGE2 groups, while the remaining groups received a solvent injection. One hour later, PGE2 (2 μL) was injected into the hypothalamus in the PEG2 and P57+PGE2 groups, while the remaining groups received a solvent injection.

[0118] 3.4 Results

[0119] In the MCAO model, we found that the body temperature of the rats in the control group was higher than that in the sham operation group after surgery. We speculated that P57 also has a cooling effect in the fever model. To this end, we implanted a double-tube micro-drug delivery system in the hypothalamus of C57BL / 6J male adult mice. Two weeks later, P57 or solvent was injected into the hypothalamus, and PGE2 was injected 1 hour later to induce fever. The core temperature changes of the mice were monitored. The results showed that although the hypothermia induced by the injection of P57 could not inhibit the fever caused by PGE2, it could keep the core temperature of the animals in a mild and sustained hypothermia state ( Figure 5 ). Therefore, P57 can be used to lower the body temperature of animals, thereby exerting central regulatory activity and protecting animal nerves.

[0120] Example 4 Effects of P57 Analogs in Treating Fever

[0121] 4.1 Materials

[0122] 4.1.1 Prostaglandin E2 (PGE2), compounds 6-70 and compounds 2-74-157.

[0123]

[0124] 4.2 Solution

[0125] 4.1.1 PGE2 solution: Dissolve PGE2 in saline to 2 mM.

[0126] 4.3 Methods:

[0127] Fever Model: Mice in good health were randomly divided into a PGE2 group and a compound-treated + PGE2 group, implanted 2 weeks prior to treatment. Mice were weighed and their body weights were recorded. Body temperatures were recorded in metabolic cages starting 1 hour before treatment. The compound-treated + PGE2 group received an intraperitoneal injection of the compound (25 mg / kg), while the PEG2 group received a solvent injection. One hour later, both groups received a hypothalamic injection of PGE2 (2 μL).

[0128] 4.4 Results

[0129] A double-tube microinjection system was implanted in the hypothalamus of adult C57BL / 6J male mice. Two weeks later, the compound of the present application or the solvent was injected into the hypothalamus. One hour later, PGE2 was injected to induce fever. The core temperature of the mice was monitored. The results showed that compared with the control group injected with the vehicle, the injection of the compound of the present application could keep the core temperature of the animals in a mild and sustained hypothermia ( Figure 6 ).

[0130] In the above examples, the compounds 6-70 and 2-74-157α used were prepared with reference to the prior art PNAS, October 7, 2014, vol. 111, No. 40, 14571–14576, and their characterization data are as follows:

[0131] Compound 6-70

[0132]

[0133] [α] D 27 =6.1(c 2.1,CHCl3); 11H NMR (400 MHz, CDCl3) δ 6.93 (q, J = 6.8 Hz, 1H), 5.41 (brs, 1H), 4.84 (d, J = 8.4 Hz, 1H), 4.76 (d, J = 7.6 Hz, 1H), 4.74 (d, J = 8.0 Hz, 1H), 4.64 (dd, J = 12.0, 4.0 Hz, 1H), 4.50 (d, J = 8.4 Hz, 1H), 4.26 (brs, 1H), 3.92 - 3.78 (m, 6H), 3.58 - 3.48 (m, 1H), 3.45 (s, 6H), 3.44 (s, 6H), 3.39 (s, 3H), 3.32 - 3.10 (m, 7H), 2.59 (brs, 1H), 2.40 - 2.26 (m, 3H), 2.20 (s, 3H), 2.14 - 2.09 (m, 1H), 1.33 - 1.12 (m, 12H), 1.06 (s, 3H), 0.98 (s, 3H); 13 13C NMR (100 MHz, CDCl3) δ 217.0, 167.6, 138.9, 137.7, 128.7, 121.9, 101.4, 99.7, 99.6, 95.8, 85.6, 82.5, 82.42, 82.38, 80.5, 77.2, 76.9, 75.8, 75.3, 71.5, 68.5, 68.3, 68.2, 58.1, 58.0, 57.9, 57.1, 56.2, 53.7, 43.0, 38.6, 37.2, 37.0, 35.6, 35.5, 35.4, 35.3, 35.2, 34.3, 33.1, 29.6, 29.4, 27.3, 26.0, 24.3, 19.2, 18.2, 18.14, 18.11, 17.9, 14.4, 12.1, 9.8; HRMS (MALDI) m / z calcd C 54 H 86 O 17 Na[M + Na] + 1029.5757, found 1029.5758.

[0134] Compound 2,74 - 157 - α:

[0135]

[0136] [α] D 25 = +56.0 (c 0.55, CHCl3); 1H NMR(400MHz,CDCl3)δ6.92(br d,J=7.6Hz,1H),5.40(br,1H),4.91(d,J=4Hz,1H),4.85(dd,J=1.6,9.6Hz,1H),4.64(dd,J=11.6,4.4Hz,1H),4.33(d,J= 7.6z,1H),4.27(s,1H),4.22(dd,J=6.8,9.2Hz,1H),3.83-3.86(m,2H),3.75(d,J=3.6Hz,1H),3.64(s,3H),3.52-3.58(m, 2H),3.40(s,3H),3.39(s,3H),3.40-3.52(m,3H),3.19(t,J=9.2Hz,1H),3.08-3.15(m,2H),2.53(s,1H),2.45(s,1H),2. 27-2.34(m,2H),2.19-2.24(m,1H)2.19(s,1H),1.88(s,3H),1.27-1.31(m,7H),1.22-1.25(m,5H),1.05(3H)0.98(s,3H); 13 C NMR (100MHz, CDCl3) δ217.1,167.7,138.0,137.8,128.7,122.0,104.2,95.7,91. 2,85.7,85.2,77.0,76.9,75.9,75.1,74.7,74.5,74.4,72.4,71.7,68.9,63.3,6 0.6,57.4,57.2,56.6,53.7,43.0,38.6,37.2,37.1,35.7,34.6,34.4,33.1,31.2 ,29.5,27.3,26.0,24.4,19.3,18.5,17.8,17.7,14.5,12.1,9.9; LR-ESIMSCalcd for C47H74O15Na(M+Na + )901.5,found 901.2.

[0137] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. Use of compound P57 or its analogues, characterized in that, For preparing a pharmaceutical composition for reducing the body temperature of a subject, wherein the compound P57 or its analogue has a structure selected from the following group:

2. The use according to claim 1, characterized in that The compound is P57(1):

3. The use according to claim 1, characterized in that The compound is 6-70 or 2,74-157-α:

4. The use according to claim 1, wherein The pharmaceutical composition is used for treating or preventing hypoxic-ischemic diseases caused by hyperthermia.

5. The use according to claim 4, characterized in that The pharmaceutical composition is used for protecting neurons in the treatment of hypoxic-ischemic diseases caused by hyperthermia.

6. The use according to claim 5, characterized in that The hypoxic-ischemic disease is selected from the group consisting of stroke, traumatic brain injury, global cerebral ischemia after cardiac arrest, myocardial ischemia, myocardial injury and hypoxic-ischemic disease.

7. The use according to claim 1, characterized in that The pharmaceutical composition is used to treat or improve fever symptoms of a subject caused by hyperthermia.

8. The use according to claim 1, characterized in that The pharmaceutical composition is an injection.

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