CN114588164B - Application of remimazolam in preventing perioperative hypothermia and chills - Google Patents

Abstract

The invention discloses the application of remimazolam in preventing perioperative hypothermia and shivering. The applicant previously found that the shivering and hypothermia after spinal canal anesthesia were significantly reduced in the remimazolam-administered group compared with the blank control group, and this application designed animal experiments to further verify. The research results of the present invention provide an effective treatment plan for reducing hypothermia in the perioperative period.

Description

Application of remimazolam in preventing perioperative hypothermia and chills

technical field

The invention belongs to the field of biomedicine and relates to the application of remimazolam in preventing perioperative hypothermia and shivering.

Background technique

Perioperative hypothermia was defined as a decrease in perioperative body temperature to less than 36.0°C. The incidence of perioperative hypothermia was 78.6%. The incidence of hypothermia was 56.6% within 2 hours and 100% after 2 hours. High American Society of Anesthesiologists (ASA) scores, 3-4 grade surgery, endoscopic surgery, anesthesia time greater than 2 hours, intravenous and irrigation fluids without warming, infusion or irrigation greater than 1000 ml significantly increased the incidence of hypothermia. Of note, a 1°C decrease in core temperature (i.e., a 2.7% decrease from normal) was associated with multiple adverse effects, including increased transfusion requirements, coagulopathy, myocardial hypoxia and arrhythmias, prolonged recovery time, immunosuppression, surgical site Increased susceptibility to infection, altered pharmacokinetics. Hypothermia can also affect the health of the patient, with the discomfort caused by chills while recovering from anesthesia comparable to postoperative pain. The consequences of hypothermia in animals are less certain due to the small number of reported studies. Cardiovascular side effects include decreased heart rate and cardiac output and prolongation of the QT interval. Body temperature is tightly regulated by hormones and cellular metabolism to maintain a stable body temperature. However, perioperative hypothermia is common secondary to anesthesia and surgical exposure. Prevention and maintenance of body temperature should start 1–2 hours before induction of anesthesia, for which both active and passive warming systems are effective in preventing perioperative hypothermia-related complications. Therefore, aggressive body temperature management before, during, and after surgery is needed to reduce the risk of perioperative hypothermia.

Prevention and treatment of perioperative hypothermia The incidence of hypothermia in perioperative patients is high, and its prevention and management need to be improved urgently. Active body temperature protection measures can reduce the incidence of hypothermia and subsequent complications in patients. According to the opinions of the expert group, the expert consensus has formulated a specific operation process for the assessment and prevention of hypothermia in perioperative patients, covering preoperative, intraoperative, and postoperative 3 stage.

Despite the widespread use of warming systems, there is a clear consensus of experts to guide them. There are still patients with hypothermia. In addition to the method of external heating, it is also very important to clarify the mechanism of perioperative hypothermia. However, the mechanism of perioperative hypothermia is still unclear. Is there any drug that can prevent and reduce the occurrence of hypothermia? rate and extent? Therefore, it is urgent to elucidate the pathogenesis of general anesthesia-induced hypothermia and find effective treatment strategies.

Contents of the invention

The applicant previously found that the shivering and hypothermia after spinal anesthesia were significantly reduced in the remimazolam administration group compared with the blank control group. Therefore, the applicant highly suspects that remimazolam may be a potential drug for the prevention and treatment of perioperative hypothermia. At present, there is no evidence from animal experiments and clinical experiments that remimazolam has the effect of treating perioperative hypothermia. Therefore, this application designs animal experiments to verify the above hypothesis, expecting to provide an effective treatment strategy for reducing perioperative hypothermia in clinic.

The invention provides the application of remimazolam in the preparation of medicine for preventing or treating perioperative hypothermia.

Further, the perioperative hypothermia is caused by general anesthesia.

Further, the general anesthesia may include administration of sevoflurane and esketamine.

Further, the medicine also includes pharmaceutically acceptable ingredients.

The invention also provides the application of remimazolam in the preparation of medicines for regulating sympathetic tension.

The present invention also provides the application of remimazolam in the preparation of reagents for regulating the expression of Ucp1 or NE.

Further, regulating the expression of Ucp1 or NE is to promote the expression of Ucp1 or NE.

Agents that promote the expression of Ucp1 or NE are referred to as Ucp1 enhancers or NE enhancers.

Ucp1 promoter refers to any material that can improve the stability of Ucp1 gene or expression product, up-regulate the expression of Ucp1, increase the effective time of gene Ucp1 or promote the transcription of Ucp1 gene. The expression of useful substances.

NE promoter refers to any material that can improve the stability of NE gene or expression product, up-regulate the expression of NE, increase the effective time of gene NE or promote the transcription of NE gene. The expression of useful substances.

As a preferred mode of the present invention, the Ucp1 promoter or NE promoter is an expression vector containing Ucp1 or NE. The expression vector usually also contains a promoter, an origin of replication and/or a marker gene and the like.

Methods well known to those skilled in the art can be used to construct the expression vector required by the present invention. These methods include in vitro recombinant DNA technology, DNA synthesis technology, in vivo recombination technology and the like. The expression vector preferably contains one or more selectable marker genes to provide phenotypic traits for selection of transformed host cells, such as karamycin, gentamicin, hygromycin, ampicillin resistance.

In the present invention, various vectors known in the art, such as commercially available vectors, include plasmids, cosmids, phages, viruses, and the like. The introduction of expression vectors into host cells can be performed by electroporation, calcium phosphate method, liposome method, DEAE dextran method, microinjection, virus infection, lipofection, and binding to cell membrane-permeable peptides. and other well-known methods.

The present invention also provides a pharmaceutical composition for preventing or treating perioperative hypothermia, the active ingredient of which is remazolam.

Further, the pharmaceutical composition also includes a pharmaceutically acceptable carrier.

The present invention also provides a method for preventing or treating perioperative hypothermia, the method comprising administering remimazolam to a subject.

Further, the subject is a patient in the perioperative period.

Furthermore, the pharmaceutical composition of the present invention also includes pharmaceutically acceptable carriers and/or adjuvants.

The pharmaceutical composition of the present invention can also be used in combination with other drugs for the prevention or treatment of perioperative hypothermia, and other therapeutic compounds can be administered simultaneously with the main active ingredients, even in the same composition.

The pharmaceutical compositions of the present invention can also be used to administer other therapeutic compounds alone, in separate compositions or dosage forms different from the main active ingredient. Some doses of the principal ingredient may be administered concurrently with other therapeutic compounds, while other doses may be administered alone. During the course of treatment, the dose of the pharmaceutical composition of the present invention can be adjusted according to the severity of symptoms, the frequency of relapses and the physiological response to the treatment regimen.

"Pharmaceutically acceptable carrier" refers to a carrier for the administration of a therapeutic agent, including various excipients and diluents. The term refers to pharmaceutical carriers which, by themselves, are not essential active ingredients and which are not unduly toxic upon administration. Suitable vectors are well known to those of ordinary skill in the art. Pharmaceutically acceptable carriers in compositions may contain liquids, such as water, saline, buffers. In addition, there may also be auxiliary substances in these carriers, such as fillers, lubricants, glidants, wetting agents or emulsifiers, pH buffering substances, and the like. The carrier may also contain cell transfection reagents.

The pharmaceutically acceptable carrier can be one or more, and the carrier includes but is not limited to diluents such as lactose, sodium chloride, glucose, urea, starch, water, etc.; binders such as starch, pregelatinized Starch, Dextrin, Maltodextrin, Sucrose, Gum Arabic, Gelatin, Methylcellulose, Carboxymethylcellulose, Ethylcellulose, Polyvinyl Alcohol, Polyethylene Glycol, Polyvinylpyrrolidone, Alginic Acid And alginate, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose, etc.; surfactants such as polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, monoglycerol stearate esters, cetyl alcohol, etc.; wetting agents such as glycerin, starch, etc.; adsorption carriers such as starch, lactose, bentonite, silica gel, kaolin and bentonite, etc.; lubricants such as zinc stearate, glyceryl monostearate , Polyethylene Glycol, Talc, Calcium and Magnesium Stearate, Macrogol, Boric Acid Powder, Hydrogenated Vegetable Oil, Sodium Stearyl Fumarate, Polyoxyethylene Monostearate, Monolauric Sucralate, Laurel Sodium alcohol sulfate, magnesium lauryl sulfate, magnesium lauryl sulfate, etc.; fillers such as mannitol (granular or powder), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugar, Coupling sugar, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate and sodium bicarbonate, etc.; disintegrants such as cross-linked vinylpyrrolidone, sodium carboxymethyl starch, low-substituted Hydroxypropyl methyl, croscarmellose sodium, soybean polysaccharide, etc.

The pharmaceutical composition of the present invention may also include additives such as stabilizers, bactericides, buffers, isotonic agents, chelating agents, pH control agents and surfactants.

Description of drawings

Figure 1 shows the results of remimazolam inhibition of hypothermia induced by sevoflurane inhalation general anesthesia; in the figure: Sevo+NS: sevoflurane+normal saline group; Sevo+RM: sevoflurane+remimazolam group ; compared with Sevo+NS: ***P<0.001, n=10, two-way ANOVA, post-hoc Tukey;

Figure 2 shows the results of remimazolam inhibiting the hypothermia induced by intravenous general anesthesia with esketamine; in the figure: Eske+NS: esketamine+normal saline group; Eske+RM: esketamine+remimazolam group . Compared with Eske+NS: ***P<0.001, n=10, two-way ANOVA, post-hoc Tukey;

Figure 3 shows the results of ELISA; A: Ucp1 expression level, B: NE expression level; in the figure: Sevo+NS: sevoflurane+saline group; Sevo+RM: sevoflurane+reimazolam group; Eske+ NS: esketamine + normal saline group; Eske+RM: esketamine + remimazolam group; compared with Sevo+NS: *P=0.0066, ***P<0.001; compared with Eske+NS: $P=0.0222,; $$$P<0.001, n=5, One-way ANOVA, post-hoc Tukey.

Detailed ways

The content of the present invention can be understood more easily by referring to the following examples, which are only for further illustrating the present invention, and are not meant to limit the scope of the present invention.

The experimental methods used in the following examples are conventional methods unless otherwise specified.

The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

The effect of embodiment remimazolam on hypothermia after general anesthesia

1. Experimental method

1. Experimental grouping

Twenty-four male SD mice, one month old, were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences of the Chinese People's Liberation Army. They were divided into 4 groups (n=6) by random number table method:

1) Sevoflurane+normal saline group (Sevo+NS group): 2.5% sevoflurane anesthesia, followed by continuous intraperitoneal infusion of normal saline 0.2ml·kg ^-1 ·h ^-1 for 60 minutes;

2) Sevoflurane+remimazolam group (Sevo+RM group): 2.5% sevoflurane anesthesia, followed by continuous intraperitoneal infusion of 0.2 mg·kg ^-1 ·h ^-1 remimazolam for 60 minutes;

3) Esketamine + normal saline group (Eske+NS): intraperitoneal injection of esketamine 100mg/kg, followed by continuous intraperitoneal infusion of normal saline 0.2ml·kg ^-1 ·h ^-1 for a total of 60min;

4) Esketamine+remimazolam group (Eske+RM): intraperitoneal injection of esketamine 100 mg/kg, followed by continuous intraperitoneal infusion of 0.2 mg·kg ^-1 ·h ^-1 remimazolam for a total of 60 min.

2. Rectal body temperature monitoring

Begin monitoring rectal body temperature immediately after anesthesia: 5min before anesthesia (-5min), immediately after the disappearance of righting reflex in mice (0min), and 5, 10, 15, 20, 60min after the start of infusion, the rectal body temperature is measured as the core temperature. The temperature in the laboratory is 22°C.

3. ELISA

Subcutaneous brown fat and plasma 0.5ml in the neck of each mouse after the last body temperature test. ELISA method was used to measure the expression of Uncoupling Protein 1 (Ucp1) protein, as well as the expression levels of norepinephrine (Norepinephrine, NE), inflammatory factors IL-1β and IL-6 in plasma. Adipose tissue was added into pre-cooled tissue protein lysate, and ground into tissue homogenate. The homogenate was centrifuged at 4°C for 5 min at 12000 rpm with a centrifugal radius of 10 cm, and the supernatant was the total protein of spinal cord tissue. Use Mouse UCP1 ELISA Kit (Colorimetric) (NOVUS Biologicals, NBP2-82438, Detection Range: 2.6-10pg/ml China), Norepinephrine (NE) ELISA Kit (BioVision, E4360-100, Detection Range: 15.625-1000pg/ml, USA) , IL-1beta Mouse Uncoated ELISA Kit (Invitrogen, Cat#88-7013-88, USA), IL-6Mouse Uncoated ELISA Kit (Invitrogen, Cat#88-7064-88, USA). The levels of Ucp1, NE, IL-1β and IL-6 were measured by experiments according to the instructions of the instructions.

4. Statistical analysis

Graph Prism 9 statistical software was used for analysis and drawing, and a box-and-whisker plot was made. Each "box" reflects the median and the 25th and 75th quartiles, and the corresponding "whiskers" represent the 5th and 75th quartiles. The 95th percentile, the measurement data of the randomized block design were compared using one-way analysis of variance, the measurement data of the repeated measurement design were compared using the analysis of variance of the repeated measurement design, and multiple groups were compared using the post-hoc Tukey test, n=10, P<0.05 difference is statistically significant.

2. Experimental results

(1) General anesthesia can cause perioperative hypothermia

Compared with 5 minutes before anesthesia, the application of sevoflurane inhalation general anesthesia or esketamine intravenous general anesthesia can cause a decrease in body temperature after infusion, which starts from 5 minutes after general anesthesia and lasts until the 20th minute. It is basically stable at 60 minutes after general anesthesia There was no statistical difference in body temperature and the first 20 minutes after anesthesia. It shows that the body temperature drops the fastest and reaches a lower level 20 minutes after general anesthesia. (All P<0.05) (Figures 1 and 2).

(2) Remimazolam inhibits hypothermia induced by sevoflurane inhalation in general anesthesia

The results are shown in Figure 1, 5 minutes after sevoflurane general anesthesia combined with remimazolam infusion (Sevo+RM), body temperature increased by 0.33℃ compared with that of sevoflurane anesthesia alone (Sevo+NS); sevoflurane general anesthesia combined with Ten minutes after remimazolam infusion (Sevo+RM), body temperature increased by 0.4°C compared with that of sevoflurane anesthesia alone (Sevo+NS); At 15 minutes, body temperature was 0.5℃ higher than that under sevoflurane anesthesia alone (Sevo+NS); NS) increased by 0.55°C; 60 minutes after sevoflurane general anesthesia combined with remimazolam infusion (Sevo+RM), body temperature increased by 0.56°C compared with sevoflurane anesthesia alone (Sevo+NS).

(3) Remimazolam inhibits hypothermia induced by intravenous general anesthesia with esketamine

The results are shown in Figure 2. Five minutes after esketamine general anesthesia combined with remimazolam infusion (Eske+RM), body temperature increased by 0.21°C compared with simple esketamine anesthesia (Eske+NS); 10 minutes after remimazolam infusion (Eske+RM), body temperature increased by 0.24°C compared with simple esketamine anesthesia (Eske+NS); At 15 minutes, body temperature was 0.25℃ higher than that of simple esketamine anesthesia (Eske+NS); NS) increased by 0.3°C; 60 minutes after esketamine general anesthesia combined with remimazolam infusion (Eske+RM), body temperature increased by 0.38°C compared with simple esketamine anesthesia (Eske+NS).

Rectal temperature monitoring confirmed the positive effect of remimazolam on hypothermia after general anesthesia, with a significant thermoprotective effect. Continuous intraperitoneal infusion of remimazolam 0.2 mg·kg ^-1 ·h ^-1 can reduce the hypothermia induced by general anesthesia.

(4) Research on the molecular mechanism of remimazolam in reducing body temperature after general anesthesia

Remimazolam may reduce hypothermia induced by general anesthesia by increasing the expression of Ucp1 and NE. Remimazolam can increase the expression of Ucp1 in brown fat of mice, suggesting that Remimazolam may increase the heat production of brown fat by increasing the expression of Ucp1, thereby increasing body temperature. In addition, studies have found that the expression of NE in brown fat increases after infusion of remimazolam, which may be through increasing sympathetic nerve tone and increasing sympathetic regulation of brown fat, thereby increasing brown fat heat production.

Although the embodiments of the present invention have been shown and described, those skilled in the art can understand that various changes, modifications, substitutions and modifications can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the invention is defined by the claims and their equivalents.

Claims (3)

1. Use of rayl Malun in the manufacture of a medicament for the prevention or treatment of perioperative hypothermia resulting from general anesthesia.

2. The use according to claim 1, wherein general anaesthesia can include administration of sevoflurane, esketamine.

3. The use according to claim 1, wherein the medicament further comprises a pharmaceutically acceptable carrier and/or adjuvant.