Anesthesia resuscitation agent as well as preparation method and application thereof

Abstract

The invention discloses an anesthesia resuscitation agent as well as a preparation method and application thereof. The anesthesia resuscitation agent comprises proadifen hydrochloride. According to the application, a novel anesthetic resuscitant can be provided, which is beneficial to rapid awakening after anesthesia, and further beneficial to improvement of anesthetic side effects, reduction of postoperative complications and improvement of anesthetic safety.

Description

Technical Field

[0001] The present application relates to the field of biomedical technology, and in particular to an anesthetic resuscitation agent for accelerating anesthesia recovery process, and a preparation method and application thereof. Background Art

[0002] General anesthesia is an indispensable part of modern surgery. By using anesthetic drugs, patients can be in a painless and memoryless state during the operation, and reflex activity can be suppressed, thereby ensuring the smooth progress of the operation. Among them, inhaled anesthetics, such as isoflurane, sevoflurane, desflurane, etc., are widely used because of their easy control and rapid metabolism.

[0003] However, the use of inhaled anesthetics may cause a series of side effects, including but not limited to respiratory depression, changes in cardiovascular function, nausea and vomiting, etc. Most of the existing anesthesia resuscitation methods rely on waiting for the natural metabolism of anesthetic drugs or the use of specific antagonists. However, these methods often have the following shortcomings: long recovery time: it takes a long time for patients to fully recover from anesthesia, increasing the risk of complications; specific antagonist limitations: not all anesthetic drugs have available antagonists, and the use of some antagonists may cause additional side effects; individual differences: the metabolic rate of anesthetic drugs varies from person to person, which makes it difficult to accurately predict the recovery time; safety: the safety and effectiveness of existing resuscitation methods need to be further improved.

[0004] Therefore, developing a new anesthetic resuscitation agent that can effectively promote anesthesia recovery is of great significance for improving anesthetic side effects, reducing the occurrence of postoperative complications, and improving anesthesia safety. Summary of the invention

[0005] The purpose of the present application is to provide an anesthesia resuscitation agent and a preparation method and application thereof.

[0006] In order to solve the above problems, this application adopts the following technical solutions:

[0007] The first aspect of the present application discloses the use of a substance in the preparation of an anesthesia resuscitation agent, wherein the substance includes prodifen hydrochloride. It should be noted that, in other words, the present application discloses the use of a substance as an anesthesia resuscitation agent, wherein the substance includes prodifen hydrochloride.

[0008] In one implementation of the present application, the anesthesia resuscitation agent is used to resuscitate the anesthetic effect produced by an inhaled anesthetic drug.

[0009] In one implementation of the present application, the inhaled anesthetic drug includes but is not limited to isoflurane.

[0010] In one implementation of the present application, the inhaled anesthetic drug includes one or more of isoflurane, sevoflurane, desflurane and enflurane. It should be noted that for other anesthetics with the same or similar anesthetic mechanism as isoflurane, the anesthesia resuscitation agent of the present application can also be used for anesthesia resuscitation.

[0011] In one implementation of the present application, the dosage form of the anesthesia resuscitation agent is in the form of powder, granules, tablets or injection. It should be noted that the powder, granules or tablets are easy to store and transport, and when used, physiological saline or other reagents can be used to prepare an anesthesia resuscitation preparation of a specific concentration; the injection can be directly used for the patient or prepared with other reagents to prepare an anesthesia resuscitation preparation of a specific concentration.

[0012] In one implementation of the present application, the anesthesia resuscitation agent is administered by intravenous injection, intramuscular injection or intraperitoneal injection.

[0013] In one implementation of the present application, the dosage of the anesthesia resuscitation agent is 10-30 mg / kg. Preferably, the dosage of the anesthesia resuscitation agent is 20 mg / kg.

[0014] The second aspect of the present application discloses the use of a substance in the preparation of a drug for activating the activity of noradrenergic neurons in the locus coeruleus or enhancing the activity of noradrenergic neurons in the locus coeruleus, and the substance includes prodifen hydrochloride. It should be noted that the present application found that prodifen hydrochloride can activate or enhance the activity of noradrenergic neurons in the locus coeruleus, and this mechanism of action can provide a treatment plan for other diseases that require activation or enhancement of the activity of noradrenergic neurons in the locus coeruleus. It should also be noted that, in other words, the present application discloses the use of a substance in activating the activity of noradrenergic neurons in the locus coeruleus or enhancing the activity of noradrenergic neurons in the locus coeruleus, and the substance includes prodifen hydrochloride.

[0015] The third aspect of the present application discloses an anesthesia resuscitation agent, which comprises prodifen hydrochloride.

[0016] In one implementation of the present application, the anesthesia resuscitation agent promotes anesthesia recovery by activating the activity of noradrenergic neurons in the locus coeruleus or enhancing the activity of noradrenergic neurons in the locus coeruleus.

[0017] The beneficial effects of this application are:

[0018] The present application can provide a new type of anesthesia resuscitation agent. Using the anesthesia resuscitation agent of the present application can effectively promote the anesthesia recovery process, facilitate rapid awakening after anesthesia, and then help improve the side effects of anesthesia, reduce the occurrence of postoperative complications, and improve the safety of anesthesia. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a schematic diagram of the experimental arrangement of anesthesia behavioral detection involved in the embodiment of the present application.

[0020] Figure 2 It is a schematic diagram of the statistical results of the anesthesia behavioral detection involved in the embodiment of the present application.

[0021] Figure 3 It is a confocal schematic diagram and statistical result diagram of the locus coeruleus c-Fos immunofluorescence staining involved in the embodiments of the present application.

[0022] Figure 4 The blue spot immunofluorescence co-staining confocal image and TH + Statistical graph of c-Fos neurons.

[0023] Figure 5 It is a schematic diagram of the detection of calcium activity in the locus coeruleus and a confocal schematic diagram involved in the embodiments of the present application.

[0024] Figure 6 It is a heat map of calcium activity of noradrenergic neurons in the locus coeruleus involved in the embodiments of the present application.

[0025] Figure 7 It is a normalized line graph and a statistical graph of the area under the line of noradrenergic neuron calcium activity involved in the embodiments of the present application.

[0026] Figure 8 It is a schematic diagram of blue spot virus injection and a schematic diagram of behavioral flow chart involved in the embodiments of the present application.

[0027] Fig. 9 This is a confocal schematic diagram of the blue spot virus injection involved in the embodiment of the present application.

[0028] Fig.10 These are the behavioral statistical results of isoflurane anesthesia involved in the examples of this application. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below by specific embodiments in conjunction with the accompanying drawings. In the following embodiments, many detailed descriptions are intended to enable the present application to be better understood. However, those skilled in the art can easily recognize that some of the features can be omitted in different situations, or can be replaced by other materials and methods. In some cases, some operations related to the present application are not shown or described in the specification, in order to avoid the core part of the present application being overwhelmed by too much description, and for those skilled in the art, it is not necessary to describe these related operations in detail, and the related operations can be fully understood according to the description in the specification and the general technical knowledge in the art.

[0030] In addition, the features, operations or characteristics described in the specification can be combined in any appropriate manner to form various implementations. At the same time, the steps or actions in the method description can also be interchanged or adjusted in a manner that is obvious to those skilled in the art. Therefore, the various sequences in the specification and the drawings are only for the purpose of clearly describing a certain embodiment and are not meant to be a required sequence, unless otherwise specified that a certain sequence must be followed.

[0031] Inhalation anesthetics are widely used in modern medicine because of their easy control and rapid metabolism. For a long time, identifying targets that promote anesthesia recovery has been a very important issue in the field of general anesthesia research, which is conducive to our development of targeted resuscitation preparations, development of new or modified existing anesthetics, and improvement of anesthesia side effects. Isoflurane (2-chloro-2-(difluoromethoxy)-l,l,l-trifluoroethane, CAS No.: 26675-46-7), as one of the inhalation anesthetics commonly used in clinical practice, is a colorless, transparent, volatile, colorless, clear liquid with a slight odor, with a relative molecular weight of 184.49 and a boiling point of 48.5°C. It is stable in nature, non-flammable and non-explosive, and easily miscible with other organic liquids. The blood / gas partition coefficient of isoflurane is lower than that of the two inhalation anesthetics, halothane and enflurane, so the rate of concentration increase in alveolar gas is faster than that of the latter two. Isoflurane is almost entirely exhaled from the lungs in its original form, and its metabolic rate in the body is less than 0.2%. It is also mainly biotransformed in the liver, where it forms metabolites such as inorganic fluoride and trifluoroacetic acid under the action of microsomal enzymes and is excreted in the urine. Isoflurane does not undergo reductive metabolism, so it does not produce free radicals. Isoflurane has a certain analgesic effect, and its effect of enhancing non-depolarizing muscle relaxants is similar to that of enflurane. Isoflurane also has the effect of dilating cerebral blood vessels and increasing intracranial pressure, but the degree is milder than that of enflurane. Isoflurane has a milder depressive effect on respiration than enflurane, and also has a bronchodilatory effect. There is no obvious damage to the liver and kidneys. In clinical practice, isoflurane is more suitable for anesthesia maintenance and is often used in surgeries that require controlled hypotension, such as ductus arteriosus occlusion and aortic stenosis.

[0032] A large number of studies have shown that the locus coeruleus (LC) in the brain plays an important role in the recovery process of isoflurane anesthesia, and the activation of noradrenergic neurons (NA) in the LC can promote recovery after isoflurane anesthesia, promote cognitive recovery, and reduce the side effects of postoperative anesthetic drugs. If there is currently a drug that can act on noradrenergic neurons in the locus coeruleus (LC NA), it will be more conducive to the rapid awakening of the majority of general anesthesia patients after anesthesia, thereby accelerating recovery. However, in current research, this type of drug is still in a state of shortage, and the development of this type of drug is of great significance for the rapid recovery of inhalation anesthesia.

[0033] In view of this, the present application creatively provides a drug for promoting anesthesia recovery, namely, an anesthesia resuscitation agent, which includes profofen hydrochloride (SKF525A). Profofen hydrochloride is also known as profofen hydrochloride or 2-(diethylamino)-2,2-diphenylpentanoic acid ethyl ester hydrochloride, with a molecular formula of C 23 H 32 ClNO2, CAS No.: 62-68-0. The present application discloses that SKF525A can effectively act on the noradrenergic neurons in the locus coeruleus of the brain after administration, accelerating the recovery after isoflurane anesthesia, which is specifically manifested in that SKF525A increases LC NA activity and accelerates the recovery of consciousness after anesthesia. The present application detected the expression of c-Fos, a marker of active neurons, in the whole brain of mice, and found that the neurons of the locus coeruleus were specifically activated by SKF525A, and the neurons activated by SKF525A were highly co-labeled with tyrosine hydroxylase (TH), a marker of noradrenergic neurons in the locus coeruleus, and after inhibiting noradrenergic neurons in the locus coeruleus by chemical genetics, the wake-promoting effect of SKF525A disappeared, thereby confirming that noradrenergic neurons in the locus coeruleus are the target of SKF525A in promoting wakefulness after isoflurane anesthesia.

[0034] This application can solve the technical problem of the lack of anesthesia resuscitation preparations for inhaled anesthetics such as isoflurane in the prior art, which is beneficial to the rapid awakening of patients under general anesthesia after anesthesia, thereby accelerating recovery, and has broad application prospects. In addition, this application also provides a drug prepared from prodifen hydrochloride that specifically activates noradrenergic neurons in the locus coeruleus, and the mechanism of action of this drug can also provide a reference for the treatment of other diseases.

[0035] The present application is further described in detail below by specific examples. The following examples are only further described for the present application and should not be construed as limiting the present application. In the following examples, unless otherwise specified, the reagents and instruments used are all commercially available, and the experimental operations are all carried out in accordance with the product instructions and conventional experimental specifications.

[0036] [Example]

[0037] Experimental subjects: 6-8 week old C57 mice were used for the experiment. All experimental animals were purchased from Guangdong Medical Animal Center and kept in the Experimental Animal Center of Southern University of Science and Technology. They were kept in the center according to standard feeding conditions, with an ambient temperature of (22±3)℃ and 12h / 12h day and night alternating light. Mice were free to eat and drink. Before the experiment, all animals were adapted to the environment for at least one week.

[0038] Anesthesia: The mice were placed in a transparent acrylic box, and isoflurane was pumped into the box at a concentration of 2% for 60 minutes, while oxygen was introduced at a flow rate of 4 L / min.

[0039] Anesthesia recovery: Prodifen hydrochloride (SKF525A) was purchased from MedChemExpress (MCE) through a domestic agent, with the item number HY-B1311. SKF525A was stored at 4°C when undissolved, and could be stored at -20°C for one month after dissolution, and at -80°C for 6 months. When used, it was dissolved in physiological saline at a concentration of 1 mg / ml by vortexing, and each mouse was intraperitoneally injected at a dose of 20 mg / kg 30 minutes before isoflurane anesthesia.

[0040] Experiment 1: Anesthesia behavioral testing

[0041] In experiment 1, mice were divided into 2 groups: the CONTROL group was intraperitoneally injected with normal saline (the injection volume was equal to the injection volume of SKF525A in the SKF525A group), and the SKF525A group was intraperitoneally injected with SKF525A, with 6 mice in each group.

[0042] Figure 1 It is a schematic diagram of the experimental arrangement of the anesthesia behavior detection involved in the embodiment of the present application. The loss of righting reflex (LORR) is widely used to judge the loss of consciousness of the anesthesia state in animal experiments. In the anesthesia behavior experiment of this embodiment, the time from LORR to the recovery of righting reflex (RORR) is called the anesthesia duration. After intraperitoneal injection of SKF525A, an isoflurane anesthesia behavior experiment is carried out. The specific steps are: C57 mice are placed in a transparent acrylic box, isoflurane is pumped into the box at a concentration of 2%, and oxygen is introduced at a flow rate of 4L / min, and then the time from LORR to RORR of the mice is recorded, which is the anesthesia duration of the mice. In order to prevent the mice from hypothermia and death during anesthesia, a heating pad is used to maintain the body temperature of the mice.

[0043] Figure 2 : is a schematic diagram of the statistical results of the anesthesia behavior test involved in the embodiment of the present application, the left side is the statistical results of the anesthesia behavior test of male mice, and the right side is the statistical results of the anesthesia behavior test of female mice. Figure 2The anesthesia behavioral results showed that in male mice, the duration of isoflurane anesthesia in the SKF525A group was significantly shorter than that in the CONTROL group (Unpaired Student t-test, **p<0.01). In female mice, the duration of isoflurane anesthesia in the SKF525A group was significantly shorter than that in the CONTROL group (Unpaired Student t-test, **p<0.05). The behavioral results between male and female individuals were consistent, so subsequent experiments were only performed for single-sex verification.

[0044] Experiment 1 shows that intraperitoneal injection of SKKF525A can accelerate recovery from isoflurane anesthesia.

[0045] Experiment 2: c-Fos immunofluorescence histochemistry to detect brain LC neuron activity

[0046] In Experiment 2, mice were divided into 2 groups: the CONTROL group was intraperitoneally injected with normal saline, and the SKF525A group was intraperitoneally injected with SKF525A, with 3 mice in each group.

[0047] The expression of c-Fos in neurons is related to the excitatory activity of neurons and is traditionally considered a sign of neuronal activation. Therefore, the changes in LC excitability after SKF525A administration can be determined by detecting the expression level of c-Fos by immunofluorescence staining. The specific method is as follows:

[0048] One hour after SKF525A injection, mice were anesthetized with isoflurane and perfused transcardially with 0.1 M PBS (phosphate buffer) followed by 4% PFA (paraformaldehyde) for tissue fixation. The brain was carefully extracted from the skull, postfixed overnight in 4% PFA at 4°C, and then transferred to 30% sucrose and dehydrated overnight at 4°C, repeated three times. The brain was cut into 30 μm coronal sections using a freezing microtome (CM 3050S, Leica). The sections were washed twice with PBS and then incubated with 5% donkey serum, 0.3% Triton-X-100 blocking solution for 60 min. The sections were then incubated overnight in the primary antibody rabbit anti-c-fos (1:500, monoclonal antibody #2250S, CST) solution. The sections were washed 3 times with PBS and then transferred to the secondary antibody Alexa Flour 488donkey anti-rabbit IgG (1:500, AB_2313584, Jackson ImmunoResearch) and incubated at room temperature for 2 h. After washing 3 times with PBS, they were transferred to DAPI (1:100000, 28718-90-3, Sigma) and incubated at room temperature for 10 min. Then, the sections were sealed with anti-fluorescence quencher (E675011-0010, BBI). Images of the stained samples were acquired and analyzed using a confocal microscope (Zeiss LSM 980) and Fiji software.

[0049] Figure 3 Schematic diagram of the confocal microscopy of c-Fos immunofluorescence staining of the locus coeruleus and the statistical results thereof are shown in FIG. Figure 3 As shown, compared with the CONTROL group, the expression level of c-Fos in the locus coeruleus of the brain in the SKF525A group was significantly increased (Unpaired Student t-test, **p<0.01), indicating that its neuronal excitability was significantly increased.

[0050] Experiment 2 showed that intraperitoneal injection of SKKF525A could increase the expression of c-Fos in locus coeruleus neurons.

[0051] Experiment 3: Immunofluorescence and histochemistry co-staining to determine neuronal types

[0052] In experiment 3, mice were divided into 2 groups: the CONTROL group was intraperitoneally injected with normal saline, and the SKF525A group was intraperitoneally injected with SKF525A, with 3 mice in each group.

[0053] Tyrosine hydroxylase (TH) is a marker for noradrenergic neurons in the LC. In order to determine that the neurons activated by SKF525A in the LC are noradrenergic neurons, c-Fos and TH were co-stained by immunofluorescence histochemistry. The specific method is as follows:

[0054] One hour after the injection of SKF525A, mice were anesthetized with isoflurane and perfused transcardially with 0.1 M PBS and then 4% paraformaldehyde (PFA) for tissue fixation. The brain was carefully extracted from the skull, postfixed in 4% PFA at 4°C overnight, and then transferred to 30% sucrose and dehydrated at 4°C overnight, which was repeated 3 times. The brain was serially cut into 30 μm coronal sections using a freezing microtome (CM 3050S, Leica). The sections were washed twice with PBS and then incubated with 5% donkey serum, 0.3% Triton-X-100 blocking solution for 60 min. The sections were then incubated overnight in primary antibody mouse anti-c-fos (1:500, ab208942, Abcam) and rabbit anti-TH (1:500, AB152, sigma) solutions. The sections were washed three times with PBS and then transferred to secondary antibodies Alexa Flour 488donkeyanti-mouse IgG (1:500, AB_2313584, Jackson ImmunoResearch) and Alexa Flour cy3donkey anti-rabbit IgG (1:500, AB_2307443, Jackson ImmunoResearch) and incubated at room temperature for 2 hours. After washing three times with PBS, they were transferred to DAPI (1:100000, 28718-90-3, Sigma) and incubated at room temperature for 10 minutes. Then, the sections were sealed with anti-fluorescence quencher (E675011-0010, BBI). Images of stained samples were acquired and analyzed using a confocal microscope (Zeiss LSM 980) and Fiji software.

[0055] Figure 4 The blue spot immunofluorescence co-staining confocal image and TH + c-Fos neuron statistics. Figure 4 As shown, after intraperitoneal injection of SKF525A, 86.89% of the increased c-Fos signals in LC were co-labeled with markers of noradrenergic neurons, demonstrating that most of the neurons activated by SKF525A were noradrenergic neurons.

[0056] Experiment 3 showed that most of the neurons activated by intraperitoneal injection of SKKF525A were noradrenergic neurons.

[0057] Experiment 4: In vivo fiber-optic calcium signal recording of noradrenergic neuron activity after SKF525A administration

[0058] In experiment 4, mice were divided into 2 groups: the CONTROL group was intraperitoneally injected with normal saline, and the SKF525A group was intraperitoneally injected with SKF525A, with 5 mice in each group.

[0059] Studies have shown that neuronal activity is closely related to the calcium ion concentration inside the neurons. When neurons discharge, a short peak of calcium ion concentration will occur. With the strict correspondence between calcium ion concentration and neural activity, neuronal activity can be reflected by calcium activity. In order to further determine the changes in the activity of noradrenergic neurons in the locus coeruleus after drug administration, in vivo fiber optic recording technology was used to record the calcium activity of noradrenaline in the locus coeruleus. The specific method is as follows:

[0060] In order to perform fiber-optic calcium signal recording of specific neurons in the locus coeruleus, rTH-Cre, Flex-GCaMP6f were injected into the locus coeruleus of adult mice, and the core needle required for fiber optic recording was implanted unilaterally in the LC. For fiber optic recording, a commercially available fiber optic photometry system (FP3001, Inper) was used. Recording was performed through a single fiber optic cable. For locus coeruleus recording, a 470nm LED was used to record GCaMP activity, and a 410nm LED was used to record the control wavelength. Data analysis of fluorescence recordings was analyzed using the software InperDataAnalysis equipped with the system, which allows the scaling of the control channel to correct for motion and bleaching, and then calculates the time histogram around the event of ΔF / F and z-score values.

[0061] Figure 5 1 is a schematic diagram of the detection of calcium activity in the locus coeruleus and a confocal schematic diagram involved in the embodiments of the present application, Figure 6 is a heat map of calcium activity of noradrenergic neurons in the locus coeruleus involved in the embodiments of the present application, Figure 7 It is a normalized line graph and a statistical graph of the area under the line of noradrenergic neuron calcium activity involved in the embodiment of the present application. Figures 5 to 7 It can be seen that after intraperitoneal injection of SKF525A, the calcium activity of noradrenergic neurons in the locus coeruleus increased significantly (Unpaired Student t-test, **p<0.01), indicating that the excitability of neurons increased significantly.

[0062] Experiment 4 showed that intraperitoneal injection of SKF525A could significantly increase the calcium activity of noradrenergic neurons in the locus coeruleus.

[0063] Experiment 5: Chemical genetics inhibition of locus coeruleus noradrenergic neuron activity for behavioral studies under isoflurane anesthesia

[0064] In experiment 5, mice were divided into 4 groups, namely, intraperitoneal injection of saline (CONTROL) group + control virus group (EGFP), intraperitoneal injection of SKF525A (SKF525A) group + specific virus group (hM4Di), intraperitoneal injection of SKF525A group + control virus group (EGFP), intraperitoneal injection of saline (CONTROL) group + specific virus group (hM4Di), with 7 mice in each group. Among them, the specific virus refers to the virus that specifically infects the norepinephrine neurons in the locus coeruleus and expresses hM4Di (specifically AAV-TH-Cre-WPRE-pA, AAV-hsyn-DIO-hM4Di-EGFP-WPRE-pA), and the control virus refers to the virus used as the blank control of the experimental group (specifically AAV-TH-Cre-WPRE-pA, AAV-hsyn-DIO-EGFP-WPRE-pA (without hM4Di)).

[0065] DREADDs (Designer Receptors Exclusively Activated by a Designer Drugs) technology is a technology that uses artificial mutations to produce G protein-coupled receptors (GPCRs) that can be specifically activated by exogenous ligands, the most commonly used being azido-N-oxide (CNO). Commonly used GPCRs include hM3Dq and hM4Di receptors. hM3Dq binds to the excitatory Gq protein to produce the excitatory hM3Dq receptor (human M3 muscarinic DREADD receptor coupled to Gq); hM4Di binds to the inhibitory Gi protein to produce the inhibitory hM4Di receptor (human M4 muscarinic DREADD receptor coupled to Gi). Exogenous administration of CNO can activate or inhibit cells expressing the above receptors, respectively. And through cell-specific promoter control, the receptor can be specifically expressed in a certain type of cell. The specific methods are as follows:

[0066] To determine whether the activation of noradrenergic neurons in the locus coeruleus is due to the arousal-promoting effect of SKF525A on isoflurane anesthesia, hM4Di was specifically expressed in noradrenergic neurons in the locus coeruleus by injecting AAV-TH-Cre-WPRE-pA virus and AAV-hsyn-hM4-DIO-EGFP-WPRE-pA virus. CNO (2 mg / kg) was injected intraperitoneally 15 minutes after intraperitoneal injection of SKF525A, and behavioral experiments under isoflurane anesthesia were performed 15 minutes later, and the duration of anesthesia was recorded.

[0067] Figure 8 is a schematic diagram of blue spot virus injection and a schematic diagram of behavioral flow chart involved in the embodiment of the present application, Fig. 9 This is a confocal schematic diagram of blue spot virus injection involved in the embodiment of the present application, Fig.10 is the behavioral statistical result of isoflurane anesthesia involved in the examples of this application. Figures 8 to 10 It can be seen that the wakefulness-promoting effect of isoflurane caused by intraperitoneal injection of SKF525A can be eliminated by specifically inhibiting LC noradrenergic neurons through chemical genetics.

[0068] Experiment 5 showed that chemical genetic inhibition of locus coeruleus noradrenergic neurons could eliminate the effect of SKF525A in promoting arousal from isoflurane anesthesia.

[0069] In summary, in this example, by performing whole-brain detection of the expression of the active neuron marker c-Fos in mice, it was found that the neurons in the locus coeruleus were specifically activated by SKF525A, and the neurons activated by SKF525A were highly co-labeled with tyrosine hydroxylase (TH), a marker of noradrenergic neurons in the locus coeruleus, and after chemical genetics inhibition of noradrenergic neurons in the locus coeruleus, the wake-promoting effect of SKF525A disappeared, thereby confirming that the noradrenergic neurons in the locus coeruleus are the target of SKF525A's action in promoting wakefulness under isoflurane anesthesia.

[0070] The above contents are further detailed descriptions of the present application in combination with specific implementation methods, and it cannot be determined that the specific implementation of the present application is limited to these descriptions. For ordinary technicians in the technical field to which the present application belongs, several simple deductions or substitutions can be made without departing from the concept of the present application.

Claims

1. Use of a substance in the preparation of an anesthetic resuscitation agent, characterized in that: The substances include prodifen hydrochloride.

2. The use according to claim 1, characterized in that: The anesthesia resuscitation agent is used to resuscitate the anesthetic effect produced by inhaled anesthetic drugs.

3. The use according to claim 2, characterized in that: The inhalation anesthetic drug includes one or more of isoflurane, sevoflurane, desflurane and enflurane.

4. The use according to claim 3, characterized in that: The dosage form of the anesthesia resuscitation agent is powder, granule, tablet or injection.

5. The use according to claim 4, characterized in that: The anesthesia resuscitation agent is administered by intravenous injection, intramuscular injection or intraperitoneal injection.

6. The use according to claim 3, characterized in that: The dosage of the anesthesia resuscitation agent is 10 to 30 mg / kg.

7. Use of a substance in the preparation of a drug for activating or enhancing the activity of noradrenergic neurons in the locus coeruleus, characterized in that: The substances include prodifen hydrochloride.

8. An anesthesia resuscitation agent, characterized in that: The anesthesia resuscitation agent includes prodifen hydrochloride.

9. The anesthesia resuscitation agent according to claim 8, characterized in that: The anesthesia resuscitation agent promotes anesthesia recovery by activating or enhancing the activity of noradrenergic neurons in the locus coeruleus.

10. A method for preparing an anesthesia resuscitation agent, characterized in that: include: Prodifen hydrochloride is dissolved in physiological saline to obtain a preparation having a predetermined concentration.

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