KR102057454B1 - Composition for inhibiting postoperative inflammatory response and composition for treating or prevanting postoperative cognitive dysfunction - Google Patents

Abstract

The present invention relates to a composition for inhibiting inflammatory response after surgery and a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery, the present invention by controlling the activation of the innate immune system and the production and secretion of proinflammatory cytokines postoperative cognitive dysfunction It provides a composition that can prevent or treat, and further inhibit the inflammatory response.

Description

A composition for suppressing inflammatory reaction after surgery and a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery {COMPOSITION FOR INHIBITING POSTOPERATIVE INFLAMMATORY RESPONSE AND COMPOSITION FOR TREATING OR PREVANTING POSTOPERATIVE COGNITIVE DYSFUNCTION}

The present invention relates to a composition for inhibiting inflammatory reaction after surgery and a pharmaceutical composition for preventing or treating postoperative cognitive dysfunction, and more specifically, a composition for inhibiting inflammatory reaction after surgery capable of suppressing the inflammatory reaction induced after surgery, and It relates to a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery.

Inflammation is a defensive reaction that occurs in the body when a living tissue is damaged. For example, it is a symptom that causes congestion, swelling, fever, and pain in a part of the body against trauma, burns, and bacterial invasion. Inflammation signals are largely produced through the cyclooxygenase (COX) pathway and the lipoxygenase (LOX) pathway, and as a result, prostaglandins, leukotriene, and thromboxane are produced. When the inflammatory signal is transmitted, various changes occur in the living body. The blood vessels in the area where inflammation is required expands to increase the blood supply, thereby supplying the blood cells necessary for the inflammatory reaction, such as neutrophils that eat bacteria.

In particular, the inflammatory response accompanying surgery may be associated with worsening the course of the disease. For example, in patients with old age, metabolic syndrome, and neurodegenerative diseases, postoperative brain inflammation may worsen the course of postoperative cognitive dysfunction (POCD). At this time, POCD, which is a cognitive decline associated with surgery, may be temporarily or lasted for several weeks to several months.In addition to prolonged hospital stay due to brain inflammation, it may lead to increased mortality, early withdrawal from work, and permanent dementia. . Accordingly, there is a need for development of methods capable of effectively preventing or treating POCD and suppressing an inflammatory response accompanying surgery.

The technology that is the background of the invention has been prepared to facilitate understanding of the present invention. It should not be understood as an admission that the matters described in the technology underlying the invention exist as prior art.

The inflammatory response can be initiated by the response of macrophages to pathogens. Inflammatory mediators such as reactive reactive oxygen species and reactive nitrogen species (e.g. NO), prostaglandins, leukotriene, etc. produced by macrophages activated by pathogens, and TNF-α, IL-6, IL-1 and IL Proinflammatory cytokines such as -8 may be involved in the inflammatory response.

Accordingly, the inventors of the present invention recognized that the activity of NF-κB, which is known as a transcription factor for genes related to the generation of inflammatory mediators, may be important for the inflammatory action of macrophages.

In addition, the inventors of the present invention recognized that the inflammatory reaction accompanying the surgery may worsen the course of the disease, and thus searched for a method of effectively suppressing it.

Accordingly, the inventors of the present invention were able to confirm that by inhibiting the transcription of NF-κB, the inflammatory response that may appear after surgery is suppressed. Furthermore, the inventors of the present invention were able to confirm the effect of improving cognitive impairment after surgery by suppressing the inflammatory response of the brain after surgery.

Accordingly, the problem to be solved by the present invention is a composition for inhibiting inflammatory response after surgery and preventing cognitive dysfunction after surgery, which can control the expression level of proinflammatory cytokines and further protein levels by inhibiting the transcription of NF-κB. It is to provide a pharmaceutical composition for use or treatment.

On the other hand, the inventors of the present invention have known that IL-1ra is an effective drug for POCD that may be associated with the inflammatory response of the brain after surgery, in particular, the specificity of the brain tissue called the blood-brain barrier (BBB). Because of this, we recognized the limitation that a dose close to 100 times the clinical dose should be used or directly administered to the ventricle.

Furthermore, the inventors of the present invention recognized the danger of an invasive method in which conventional drugs for inhibiting brain inflammation are used in an invasive amount or administered directly into the brain due to insignificant penetration of brain tissue.

Accordingly, another problem to be solved by the present invention is to provide a composition for inhibiting inflammatory reaction after surgery and a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery, in which the dosage or administration method can be practically applied clinically.

The problems of the present invention are not limited to the problems mentioned above, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

According to an embodiment of the present invention, there is provided a composition for inhibiting postoperative inflammatory reactions comprising a fusion protein comprising a transcriptional regulatory domain and a protein transport domain of NF-κB.

As used herein, the term "inflammatory reaction" may mean an inflammatory reaction accompanying surgery, but is not limited thereto. For example, the inflammatory reaction may be an inflammatory reaction induced by tissue damage caused by an autoimmune reaction.

Inflammatory reactions can contribute to restoring normal structure and function by neutralizing or eliminating pathogenic factors and regenerating damaged tissues in normal individuals, but otherwise can cause diseases such as chronic inflammation. In particular, the inflammatory response of the brain induced through immune-to-brain communication in a situation in which the systemic immune system such as infection or surgery is stimulated may be associated with diseases such as POCD.

Specifically, a series of innate immune responses and inflammatory responses caused by tissue damage by surgery proceed as follows. First, damage-associated molecular patterns (DAMPs) represented by high-mobility group box 1 (HMGB1) are secreted. Then, DAMPs or Pathogen-associated molecular patterns (PAMPs) are bound to pattern recognition receptors (PRRs) typified by TLRs. This activates nuclear factor-κB (NF-κB), thereby promoting the synthesis and secretion of pro-inflammatory cytokines. As a result, macrophages are more activated, and proinflammatory cytokines such as IL-1 and TNF-α in the damaged tissue are increased, resulting in an inflammatory response.

In particular, IL-1β not only activates various inflammatory signals, but also affects the function of the central nervous system and is expressed in various diseases, so it can be considered as a prototype of cytokines. Thus, IL-1β may be a major modulator of cytokines associated with inflammatory responses that may accompany post-surgery. For example, when IL-1β is administered or the internal IL-1β level increases due to infection or surgery, an inflammatory reaction in the tissue may occur.

Furthermore, IL-1 phosphorylates IκBα through TNF receptor-associated factor 6 (TRAF6), MyD88, and IRAK, thereby increasing the DNA-binding activity of NF-κB. This activated NF-κB amplifies IL-1, IL-6, and TNFα by transcribing inflammatory enzymes and cytokines. Accordingly, NF-κB may be closely associated with an inflammatory response accompanying surgery, and further, a disease induced by an inflammatory response. For example, an increase in the activity or expression of NF-κB can lead to degenerative brain diseases including Alzheimer's disease, which is induced by an inflammatory response in the brain. Furthermore, an increase in the activity or expression of NF-κB can cause damage to the spleen, which plays an important role in helping the function of immune cells. Specifically, the function of the spleen removes particles from the bloodstream, such as cells surrounded by bacteria or antibodies bound to opsonin, which helps the function of immune cells. Accordingly, abnormalities in the spleen may lead to the onset of systemic inflammation and sepsis by lowering the ability to fight infections and lower immunity.

Accordingly, the inventors of the present invention recognized that inhibition of NF-κB can reduce the inflammatory response, and found that NF-κB is a key regulator of inflammatory cytokines associated with various immune responses that may occur after surgery.

Furthermore, the inventors of the present invention can control the expression level of proinflammatory cytokines or the level of protein by inhibiting NF-κB transcription by competitive inhibition, and as a result of this, a composition capable of suppressing the inflammatory response accompanying surgery. Came to invent.

Accordingly, the fusion protein used in the composition for suppressing the inflammatory response after surgery according to an embodiment of the present invention may include a transcriptional regulatory domain and a protein transport domain of NF-κB. That is, the fusion protein provided in the present invention can suppress the inflammatory reaction accompanying the surgery.

As used herein, the term “transcription control domain” refers to a domain constituting a transcription factor, and does not have a transactivation domain, and refers to a domain composed of only a DNA binding site, wherein the fusion protein does not have a transactivation domain, but has a DNA binding site. The fusion protein of the present invention is a dominant negative mutant for the NF-κB gene, so it can bind to the desired promoter, but cannot promote transcription. It can act as a competitive inhibitor to inhibit the transcription and activity of NF-κB.

According to an embodiment of the present invention, NF-κB may be at least one of RelA (p65), c-Rel, Rel-B, NF-κB1 (p50) and NF-κB2 (p52), and in various embodiments NF-κB may be RelA(p65).

The term “protein transport domain” as used herein refers to a short peptide with strong hydrophobicity consisting of 7-50 amino acids, and refers to a domain capable of delivering not only proteins with a molecular weight of 120 kDa or more, but also DNA or RNA into cells. In the case of the protein to which the protein transport domain of the present invention is not attached (i.e., p65-TMD consisting of only the transcriptional regulatory domain), unlike the fusion protein of the present invention, transcriptional inhibition of NF-κB and IL-2, pro-inflammatory cytokine by LPS It has no secretion inhibitory effect.

According to an embodiment of the present invention, the protein transport domain is Hph-1, Mph-1, Sim-2, Tat, VP22, Antp (antennapedia), Pep-1 (peptide-1), PTD-5 (protein transduction domain). -5), 11R, 7R, and CTP (cytoplasmic transduction peptide) may be at least one. Here, “11R” and “7R” mean a peptide consisting of 11 and 7 arginine, respectively. In various embodiments, the protein transport domain may be Hph-1.

The inventors of the present invention have recognized that among various inflammatory reactions that can occur after surgery, the inflammatory reaction of the brain can be a mechanism of POCD. Accordingly, the inventors of the present invention confirmed the brain inflammation pattern in which proinflammatory cytokines increased in cerebrospinal fluid immediately after peripheral surgery, and furthermore, POCD occurrence showed a significant relationship with IL-6 or brain cell damage. Was confirmed.

As a result, the inventors of the present invention can control the expression level of proinflammatory cytokines or the level of protein by inhibiting NF-κB transcription by competitive inhibition, and as a result, cognitive dysfunction after surgery that can inhibit brain inflammation. A pharmaceutical composition for prophylaxis or treatment has been invented.

Accordingly, according to an embodiment of the present invention, there is provided a composition for preventing or treating cognitive dysfunction after surgery comprising a fusion protein comprising a transcriptional regulatory domain and a protein transport domain of NF-κB.

As used herein, the term "prevention" refers to any action of inhibiting or delaying cognitive dysfunction after surgery by administering the pharmaceutical composition for prophylaxis or treatment according to an embodiment of the present invention to an individual.

As used herein, the term "treatment" refers to any action to improve or benefit from symptoms of cognitive dysfunction after surgery by administering the pharmaceutical composition for prophylaxis or treatment according to an embodiment of the present invention to an individual.

As used herein, the term "postoperative cognitive dysfunction" may be the same as POCD. Postoperative cognitive dysfunction includes all symptoms that appear in the form of postoperative cognitive decline, for example, from temporary cognitive decline associated with surgery, to persistent cognitive decline for weeks to months, or dementia. It can mean not only the cognitive impairment it causes, but any condition that is likely to cause deterioration or delirium, or that needs to be prevented.

In particular, brain inflammation induced through immune-to-brain communication in a situation where the systemic immune system is stimulated such as infection or surgery may be associated with diseases such as POCD. Here, immune-to-brain communication means that cytokines in the blood reach the brain through diffusion, cytokines are transmitted to the brain through interactions between cytokines and vascular endothelial cells in the brain, or the catecholamine circuit of the vagus nerve and the sympathetic nervous system. It means that cytokines in the periphery directly activate the nervous system.

In addition, Immune-to-brain communication induces brain inflammation, and short-term brain inflammation induces central nerve-mediated pathological behaviors such as fever, decreased appetite, drowsiness, and cognitive impairment. This is considered a host's self-defense mechanism that allows the injured individual to heal itself. After healing, inflammation is weakened, and pathological behaviors including cognitive dysfunction are known to gradually disappear.

However, in the case of aged experimental animals, the duration and extent of pathological behavior tend to be much more severe than the extent of tissue damage. Returning to normal life may be delayed or practically impossible. In an environment that changes the immune response, such as aging, stress, metabolic syndrome, and neurodegenerative diseases, maladaptive pathological behavior may result, leading to pathological conditions of behavioral and cognitive function. In addition, there are many cases of showing an increased inflammatory profile even in the brain of relatively normal people without disease.

Specifically, damage to the brain tissue caused by surgery induces the activation of NF-κB, thereby promoting the synthesis and secretion of proinflammatory cytokines in the brain. In particular, TNF-α damages the blood-brain barrier, and activated macrophages derived from the bone marrow can penetrate into the brain parenchyma through the damaged blood-brain barrier. As a result, macrophages and microglia are continuously activated in the central nervous system. Thereafter, proinflammatory cytokines such as IL-1β or TNF-α in the hippocampus are increased, and the increase of proinflammatory cytokines may cause brain inflammation, leading to impaired synaptic transmission and cognitive function abnormalities.

The fusion protein provided in the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention may suppress NF-κB transcription by competitive inhibition, thereby suppressing neuroinflammation.

For example, the fusion protein is an intracellular adhesion molecule (ICM), a vascular cell adhesion molecule (VCAM), E-selectin, and P-selectin. And by suppressing the transcription of neutrophils, brain inflammation can be suppressed. At this time, the intercellular adhesion molecule, the vascular cell adhesion molecule, the E-selectin, and the P-selectin may be adhesion molecules. The adhesion molecule may be involved in the infiltration of white blood cells into the brain parenchyma. Furthermore, blood-derived leukocytes, such as neutrophils, can induce memory loss and neuropathological changes associated with Alzheimer's disease. Furthermore, neutrophilic leukocytes induced during the inflammatory response can activate microglia and disrupt tissue. Accordingly, a decrease in the expression level of intercellular adhesion molecule, vascular cell adhesion molecule, E-selectin and P-selectin, and neutrophilic leukocytes may induce inhibition of brain inflammation.

In the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention, the transcriptional regulatory domain of NF-κB is RelA (p65), c-Rel, Rel-B, NF-κB1 (p50) and NF- At least one of κB2 (p52), the protein transport domain is Hph-1, Mph-1, Sim-2, Tat, VP22, Antp (antennapedia), Pep-1 (peptide-1), PTD-5 (protein transduction domain) -5), 11R, 7R, and may be at least one of CTP (cytoplasmic transduction peptide).

The composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention may include 0.0001% to 10% by weight of the fusion protein based on the total weight of the composition. . However, it is not limited thereto and may be included in various amounts.

The transcriptional control domain of NF-κB of the composition for suppressing the postoperative inflammatory response and the composition for preventing or treating postoperative cognitive dysfunction according to an embodiment of the present invention may have an amino acid sequence of SEQ ID NO: 1, and a protein transport domain May have the amino acid sequence of SEQ ID NO: 2.

Furthermore, the fusion protein of the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention has an amino acid sequence of SEQ ID NO: 3.

The fusion protein of the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention is a deletion, insertion, non-conservative or conservative substitution of the domain containing It includes a variant of a protein by a combination of. Substitutions, insertions, deletions of the listed amino acid sequences may also be used in compositions for inhibiting inflammatory reactions after surgery and compositions for preventing or treating cognitive dysfunction after surgery in various embodiments provided by the present invention.

In this case, the protein variant refers to a protein having a sequence different from the listed amino acid sequence and one or more amino acid residues. Insertions typically consist of a contiguous sequence of about 1 to 20 amino acids, although larger insertions may be possible. Deletion usually consists of about 1 to 30 residues, but larger deletions may also be possible, such as in some cases one of the domains may be deleted. Such variants can be prepared by chemical peptide synthesis methods known in the art or recombinant methods based on DNA sequences (Sambrook et al., Molecular Cloning, Cold Spring Harbor Laboratory Press, New York, USA, 2d Ed. , 1989). Amino acid exchanges in proteins and peptides that do not totally alter the activity of the molecule are known in the art (H. Neuroth, R.L. Hill, The Proteins, Academic Press, New York, 1979). The most commonly occurring exchanges are amino acid residues Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Thy/Phe, Ala/ It is an exchange between Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu, Asp/Gly.

In addition, the fusion protein in various embodiments of the present invention is modified by phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc., depending on the case. It can also be (modification).

The variant or modified product may be a functional equivalent exhibiting the same biological activity as the fusion protein, but, if necessary, a variant or modified product in which the properties of the protein are modified. Preferably, it may be a protein having increased structural stability against heat, pH, or the like, or increased protein activity due to mutations and modifications in the amino acid sequence.

The administration route and administration method of the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention may be independent, and the method is not particularly limited, and the purpose Any route of administration and mode of administration may be followed as long as the pharmaceutical composition can reach the corresponding site. In this case, the target site may be any site that may accompany an inflammatory reaction after surgery, or any site in which a disease appears due to an inflammatory reaction after surgery. In addition, the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery according to an embodiment of the present invention may be administered orally, parenterally, or intranasally, and intranasal administration method Can also be administered.

In particular, in order to prevent cognitive dysfunction after surgery, the composition for inhibiting inflammatory reactions after surgery and the composition for preventing or treating cognitive dysfunction after surgery of the present application are preferably introduced into the central or peripheral nerves through an appropriate route. Here, an appropriate route of administration may be intranasal administration. More specifically, the route of administration includes intraventricular or intrathecal administration via intranasal administration. In this case, intranasal administration may be a non-invasive and efficient method capable of administering peptides or proteins to the brain parenchyma while reducing systemic exposure without crossing the blood-brain barrier.

For example, when the composition for inhibiting the postoperative inflammatory reaction of the present invention and the composition for preventing or treating postoperative cognitive dysfunction are administered intranasally, the composition is distributed in the olfactory nerve and the trigeminal nerve. It comes into contact with the nasal mucosa. Then, the composition for suppressing the inflammatory response after this surgery reaching the olfactory epithelium region and the composition for preventing or treating postoperative cognitive dysfunction are the epithelium barrier around the olfactory receptor neuron (ORN). These open gaps move along the periphery of the ORN axon, and can quickly enter the central nervous system through channels by the olfactory sheathing cells. At this time, the composition for suppressing the inflammatory response after surgery and some of the composition for preventing or treating cognitive dysfunction after surgery may enter the ORN through endocytosis at the end of the ORN and move to the olfactory bulb. The composition, which has not reached the olfactory epithelial region, can also be taken up intracellularly by the trigeminal nerve at the terminal of the trigeminal nerve and can be moved directly to the caudal region of the brain. In addition, the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery, which is absorbed to the proprioceptive plate even without passing through the nerve periphery, are the central nervous system , CNS). The composition, which has entered the CNS by any route, can spread throughout the brain via interstitial fluid or cerebrospinal fluid. However, as described above, the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery of the present invention are not limited to being delivered to the brain, and may be delivered to a target site where the inflammatory reaction occurs after surgery. Furthermore, the route of administration for the effective administration of the composition for inhibiting the postoperative inflammatory reaction of the present invention and the composition for preventing or treating cognitive dysfunction after surgery may be easily selected by those skilled in the art.

The composition for suppressing the postoperative inflammatory reaction of the present application and the composition for preventing or treating postoperative cognitive dysfunction may include a nasal acceptable carrier for intranasal administration, wherein the carrier is a mammal, preferably a human It refers to one or more suitable solid or filler diluents or encapsulating materials suitable for administration to any part of the nasal epithelium. Typically, the carrier may be a liquid, solution, suspension, gel, ointment, lotion, or a combination thereof. Preferably, the carrier is a pharmaceutically acceptable aqueous carrier. The composition for suppressing the postoperative inflammatory reaction of the present invention and the composition for preventing or treating cognitive dysfunction after surgery may be prepared in various unit dosage forms. Such forms include, but are not limited to, nasal drops, nasal sprays, nasal gels, nasal ointments, and nasal powders.

In addition, the carrier may include a delivery enhancer, and the intranasal delivery enhancer includes an agglutination inhibitor, a dosage modifier, a pH control agent, a degrading enzyme inhibitor, a mucolytic or mucolytic agent, a ciliated stabilizing agent, a membrane permeation accelerator (e.g. For example, surfactants, bile salts, phospholipids or fatty acid additives, mixed micelles, liposomes, or carriers, alcohols, enamines, nitric oxide donor mixtures, long-chain amphiphilic molecules, small Hydrophobic penetration enhancers, sodium or salicylic acid derivatives, glycerol esters of acetoacetic acid, cyclodextrin or beta-cyclodextrin derivatives, medium chain fatty acids, chelating reagents, amino acids or salts thereof, N-acetylamino acids or salts thereof, degradation to selected membrane components Enzymes, fatty acid synthesis inhibitors, cholesterol synthesis inhibitors or nitric oxide stimulating substances, chitosan, and agents for modulating epithelial conjugation physiology such as chitosan derivatives, vasodilators, selective delivery promoters, and to enhance intranasal mucosal delivery, the compositions of the present invention are effectively combined. Stable carriers, carriers, support materials or complex-forming species, etc., which are capable of being, bound, stored, encapsulated or stabilized in the active agent, may be included.

In the present specification, "individual" refers to all animals including mammals, including mice, livestock, and humans.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention should not be construed as being limited by these examples.

The present invention exhibits the effect of suppressing the inflammatory response that appears after surgery by controlling the activation of the innate immune system and the production and secretion of proinflammatory cytokines.

In addition, the present invention can provide a composition for inhibiting inflammatory reaction after surgery, and a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery, in which the dosage or administration method can be actually applied clinically.

Furthermore, the present invention establishes a causal relationship with brain inflammation centered on IL-1β and NF-κB, and has the potential of clinical application that can prevent POCD by blocking IL-1β and NF-κB mechanisms. Is provided.

The effects according to the present invention are not limited by the contents exemplified above, and more various effects are included in the present specification.

1A and 1B show the expression levels of IL-1β and IL-6 in the hippocampus (hippocampus) after treatment with the composition for inhibiting the inflammatory response after surgery according to an embodiment of the present invention.
1C shows the levels of M1 phenotypic cytokines and M2 phenotypic cytokines in the hippocampus after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention.
1D shows a change in the integrity of the blood-brain barrier after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention.
1E shows changes in activation of microglia in the brain after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention.
Figure 1f shows the expression level of the adhesion molecule in the hippocampus after treating the composition for inhibiting inflammatory reaction after surgery according to an embodiment of the present invention.
1G shows the expression level of neutrophils in the brain after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention.
Figure 2a shows the expression level of cytokines in the spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention.
2B shows the levels of the M1 phenotypic cytokine and the M2 phenotypic cytokine in the spleen after treatment with the composition for inhibiting the inflammatory response after surgery according to an embodiment of the present invention.
Figure 2c shows the protein levels of IL-1β, IL-6, TNFα and MCP-1 in the LPS-treated spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention.
Figure 2d shows the relative expression levels of IL-1β, IL-6, TNFα and IL-17A in the LPS-treated spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention.
Figure 3a shows the experimental results of evaluating learning and memory through a passive avoidance test after treating a pharmaceutical composition for preventing or treating cognitive dysfunction according to an embodiment of the present invention in an abdominal surgery-like model.
Figure 3b shows the experimental results of evaluating working memory through EPM (Elevated plus maze) after treating a pharmaceutical composition for preventing or treating cognitive dysfunction according to an embodiment of the present invention in an abdominal surgery-like model.
3C shows the experimental results of evaluating cognitive memory through a cognitive test for a new object after treating a pharmaceutical composition for preventing or treating cognitive dysfunction according to an embodiment of the present invention in an abdominal surgery-like model.

Advantages and features of the present invention, and a method of achieving them will become apparent with reference to the embodiments described below in detail together with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, only the present embodiments are intended to complete the disclosure of the present invention, and common knowledge in the technical field to which the present invention pertains. It is provided to completely inform the scope of the invention to those who have, and the invention is only defined by the scope of the claims.

Hereinafter, the effects of the composition for suppressing the postoperative inflammatory reaction of the present invention and the composition for preventing or treating cognitive dysfunction after surgery are illustratively described for verification of the brain or spleen, but are not limited thereto, and various The composition in the embodiment may be provided to a site where an inflammatory reaction may occur after surgery, and further to a site where a disease occurs due to an inflammatory reaction after surgery.

In the following, for simplicity of explanation, the composition for inhibiting inflammatory reaction after surgery and the composition for preventing or treating cognitive dysfunction after surgery are the composition of the present invention (nt-p65-TMD fusion protein) or the composition of the present invention. appear. In addition, the composition for inhibiting inflammatory reaction after surgery of the present invention exhibits an effect of preventing or treating POCD by inhibiting brain inflammation, and thus may be provided as a composition for preventing or treating cognitive dysfunction after surgery. Furthermore, the composition for preventing or treating cognitive dysfunction after surgery of the present invention may be provided as a composition for suppressing inflammatory reactions after surgery.

Example 1: Brain inflammatory response test

Brain inflammatory response analysis

Systemic inflammation caused by tissue damage caused by surgery causes invasion of macrophages in the brain, especially hippocampus, and activation of microglia, which is an immune cell of the nervous system, and thus NF-kB, IL-1β or TNF-α. It can keep levels of the same pro-inflammatory cytokines high. In addition, levels of anti-inflammatory cytokines such as arginase-1 and IL-10 may be lowered. High levels of proinflammatory cytokines cause cognitive dysfunction. Therefore, the innate immune system is activated by tissue damage caused by surgery, which can lead to brain inflammation.

Therefore, it was confirmed that the novel fusion protein of the present invention is effective in inhibiting brain inflammation.

1A and 1B show the expression level of cytokines in the hippocampus after treatment with a composition for inhibiting inflammatory reaction after surgery according to an embodiment of the present invention. 1A shows the results of ELISA examination by extracting the hippocampus on the second day after surgery, and confirming that IL-1β and IL-6 are effectively reduced in the hippocampus after surgery when the composition of the present invention (nt-p65-TMD fusion protein) is administered. I can. Figure 1b shows the results of the RT-PCR test by extracting the hippocampus on the second day after surgery, and IL-1β, IL-6 and TNF-α in the hippocampus after surgery when the composition of the present invention (nt-p65-TMD fusion protein) is administered. It can be seen that is effectively reduced. Furthermore, the nt-p65-TMD (5mg/kg) group treated with the composition of the present invention (nt-p65-TMD fusion protein) showed fine expression levels of anti-inflammatory cytokines such as Arginase-1 and IL-10. It can be seen that it has increased significantly. At this time, IL-1β, IL-6 and TNF-α of pro-inflammatory cytokines may be of the M1 phenotype, and Arginase-1 and IL-10 of anti-inflammatory cytokines may be of the M2 phenotype. Accordingly, the composition of the present invention (nt-p65-TMD fusion protein) may be more effective against cytokines of the M1 phenotype.

1C shows the levels of M1 phenotypic cytokines and M2 phenotypic cytokines in the hippocampus after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention. Specifically, the level of cytokines in the hippocampus was measured by extracting the hippocampus on the 2nd day after surgery, staining CD86+ cells, which are markers of the M1 phenotype, and CD206+ cells, which are markers of the M2 phenotype, and then perform flow cytometry. Figure 1c (a) shows the level of the M1 phenotypic cytokine in the hippocampus. As a result, it can be seen that the nt-p65-TMD (10mg/kg) group treated with the composition of the present invention decreased the number of CD86+ cells in contrast to the POCD group, resulting in a decrease in the level of the M1 phenotype cytokine. Furthermore, (b) of FIG. 1C shows the level of the M2 phenotypic cytokine in the hippocampus. It can be seen that the nt-p65-TMD (10 mg/kg) group treated with the composition of the present invention has a similar number of CD206+ cells to the POCD group.

Changes in the integrity of the blood-brain barrier

Surgical surgery can lead to an increase in proinflammatory cytokines produced and secreted by peripheral cells. Such systemic inflammation affects the dysfunction and integrity of the blood-brain-barrier, and leukocytes and neutrophils present in the blood penetrate into the brain tissue.

Therefore, it was confirmed that the new fusion protein of the present invention can regulate the amount of circulating blood in the blood-brain-barrier.

1D shows a change in the integrity of the blood-brain barrier after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention. Figure 1d (a) and (b) using the Evans blue extravasation method (Evans blue extravasation method) to observe the integrity in the striatum, cortex, and hippocampus on the 2nd day after surgery. This is the result of the test. At this time, the integrity was observed by administering 0.5% Evans Cheong to the jugular vein of the rat on the 2nd day after surgery, and then observing the new striae, cortex, and hippocampus. According to the results, it can be seen that the influx of Evans blue in the new striae, cortex, and hippocampus was significantly increased in the POCD group. In contrast to the POCD group, the composition (nt-p65-TMD fusion protein) administration group of the present invention significantly reduced the amount of milk of Evans Cheong. In particular, it can be seen that the nt-p65-TMD (10mg/kg) group treated with the composition of the present invention has no difference from the results of the normal control group.

1E shows changes in activation of microglia in the brain after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention. Figure 1e (a) is a result of examining the protein level of the microglia marker in the hippocampus on the 2nd day after surgery using western blot analysis. 1e (b) is a result showing the relative optical density with respect to the result of FIG. 1e (a). According to the results, it can be seen that the level of protein of microglia markers was significantly increased in the POCD group, which performed only surgery, in contrast to the control group. The composition (nt-p65-TMD fusion protein) administration group of the present invention can be confirmed that the protein level of the microglia marker decreased to a level similar to that of the control group in contrast to the POCD group. Furthermore, (c) of FIG. 1e is a result of examining the expression level of microglia in the hippocampus on the second day after surgery using immunofluorescence staining. According to the results, it can be seen that the administration group of the composition (nt-p65-TMD fusion protein) of the present invention decreased the expression level of microglia to a level similar to that of the control group in contrast to the POCD group. Microglia can produce proinflammatory cytokines as they are activated. As a result of the above, the composition of the present invention exhibits an effect of reducing the expression level of microglia or the protein level of its marker, thereby reducing the level of proinflammatory cytokines. It has a controllable effect.

Decreased expression level of adhesion molecules in vascular endothelial cells

Vascular inflammatory reactions and resulting blood-brain-barrier dysfunction can be an important pathogen for several degenerative brain diseases, including Alzheimer's disease. Furthermore, the vascular inflammatory response may mediate the invasion of blood-derived leukocytes such as lymphocytes, mononuclear leukocytes, and neutrophil leukocytes into the ventricle in the Alzheimer's disease animal model as well as the postoperative cognitive impairment animal model.

At this time, the expression of adhesion molecules in vascular endothelial cells may be involved in infiltrating the blood-derived leukocytes into the brain parenchyma. Accordingly, it was confirmed that the novel fusion protein of the present invention can regulate the expression of adhesion molecules.

Figure 1f shows the expression level of the adhesion molecule in the hippocampus after treating the composition for inhibiting the inflammatory response after surgery according to an embodiment of the present invention. 1F is an intracellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), E-selectin, and P-selectin in the hippocampus on the second day after surgery. ) Is the result of testing the mRNA level. According to the results, the composition (nt-p65-TMD fusion protein) administered group of the present invention has a similar level of the control group compared to the POCD group, and the transcription of the intercellular adhesion molecule, vascular cell adhesion molecule, E-selectin and P-selectin is similar to that of the control group. It can be seen that it is effectively reduced.

Decreased expression level of neutrophils in vascular endothelial cells

Blood-derived leukocytes such as lymphocytes, monocytes and neutrophils can induce memory loss and neuropathological changes associated with Alzheimer's disease. Furthermore, neutrophilic leukocytes induced during the inflammatory response can activate microglia and disrupt tissue.

Accordingly, it was confirmed that the novel fusion protein of the present invention can reduce the expression level of neutrophils.

1G shows the expression level of neutrophils in the brain after treatment with a composition for inhibiting inflammatory reactions after surgery according to an embodiment of the present invention. Figure 1g (a) is a result of testing the protein level of neutrophil elastase in the hippocampus on the 2nd day after surgery using western blot analysis. (B) of 1g is a result showing the relative optical density with respect to the result of (a) of FIG. 1g. According to the results, it can be seen that the level of protein of neutrophil elastase was significantly increased in the POCD group, which performed only surgery, in contrast to the control group. It can be seen that the composition (nt-p65-TMD fusion protein) administration group of the present invention decreased the protein level of neutrophil elastase to a level similar to that of the control group in contrast to the POCD group. In particular, it can be seen that the nt-p65-TMD (10mg/kg) group treated with the composition of the present invention decreased the protein level of neutrophil elastase compared to the normal control group. Further, (b) of FIG. 1G is a result of examining the expression level of neutrophil elastin in the hippocampus on the second day after surgery using immunofluorescence staining. According to the results, it can be seen that the composition (nt-p65-TMD fusion protein) administration group of the present invention reduced the expression of neutrophil elastin at a level similar to that of the control group in contrast to the POCD group. The increase in neutrophilic leukocyte activity induced by the inflammatory response can activate microglia cells that produce proinflammatory cytokines. As a result of the above, the composition of the present invention exhibits an effect of reducing the expression level of neutrophil elastatis or the level of neutrophil elastatis protein, thereby controlling the level of pro-inflammatory cytokines.

Example 2: Spleen inflammatory response test

Surgery damage to the spleen can increase the level of pro-inflammatory cytokines such as NF-kB, IL-1β, or TNF-α. As a result of this, the inflammatory response can be systemic.

Therefore, it was confirmed that the novel fusion protein of the present invention can suppress the inflammatory response in the spleen.

Analysis of the inflammatory response of the spleen (inflammation response model performed by surgery)

Figure 2a shows the expression level of cytokines in the spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention. Figure 2a shows the results of the RT-PCR test by extracting the hippocampus on the 2nd day after surgery, and IL-1β, IL-6 and TNF-α in the hippocampus after surgery when the composition of the present invention (nt-p65-TMD fusion protein) is administered. It can be seen that is effectively reduced. Furthermore, it can be seen that the nt-p65-TMD (5mg/kg) group treated with the composition of the present invention increased the expression level of anti-inflammatory cytokines such as arginase-1 and IL-10. In particular, the relative expression level of IL-10 in the nt-p65-TMD (5mg/kg) group treated with the composition of the present invention showed similar results to the control group, and thus the composition of the present invention was used to control the expression level of IL-10. You can see that this is effective. Accordingly, the composition of the present invention (nt-p65-TMD fusion protein) can more effectively regulate the expression levels of IL-1β, IL-6 and TNF-α of the M1 phenotype, and IL-10, which is the M2 phenotype, than other cytokines.

2B shows the levels of the M1 phenotypic cytokine and the M2 phenotypic cytokine in the spleen after treatment with the composition for inhibiting the inflammatory response after surgery according to an embodiment of the present invention. Specifically, the level of cytokine in the spleen was measured by extracting the spleen on the second day after surgery, staining CD86+ cells, a marker of the M1 phenotype, and CD206+ cells, a marker of the M2 phenotype, and then performing flow cytometry. Figure 2 (a) shows the level of the M1 phenotypic cytokine in the spleen. As a result, it can be seen that the number of CD86+ cells in the nt-p65-TMD (10 mg/kg) group treated with the composition of the present invention decreased in contrast to the POCD group. Furthermore, it can be seen that the level of the M1 phenotypic cytokine in the nt-p65-TMD (10mg/kg) group was reduced to a level similar to that of the control group. Figure 2b (b) shows the level of the M2 phenotypic cytokine in the spleen. It can be seen that the nt-p65-TMD (10 mg/kg) group treated with the composition of the present invention has the number of CD206+ cells similar to that of the control group in contrast to the POCD group. The results of FIG. 2B are also linked to the result that the expression level of the M1 phenotype of the inflammatory cytokine in FIG. 2A decreased similarly to the level of the control group, and the expression level of the M2 phenotype of the anti-inflammatory cytokine increased similarly to the level of the control group. do.

Analysis of Spleen's Inflammatory Response (LPS-induced Inflammatory Response Model)

It was confirmed that the new fusion protein of the present invention is effective not only for the inflammatory response induced by surgery, but also for the inflammatory response induced by LPS. Hereinafter, the effect of the novel fusion protein of the present invention on the LPS-treated spleen is exemplarily described, but the effect thereof is not limited to the spleen.

At this time, as the inflammatory response model, BALB/c mice (6 to 8 weeks) to which LPS (20 mg/kg) was administered once intraperitoneally were used.

Figure 2c shows the protein levels of IL-1β, IL-6, TNFα and MCP-1 in the LPS-treated spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention. 2C shows the results of ELISA test by extracting the spleen on the 2nd day after LPS administration. Specifically, the levels of inflammatory cytokines such as IL-1β, IL-6, TNFα and MCP-1 were increased in the LPS-treated group, and in the case of the group administered with the composition of the present invention (nt-p65-TMD fusion protein), LPS Compared to the treatment group, the protein level of inflammatory cytokines was decreased. Furthermore, compared to the normal control group, there was no significant difference between the group administered the composition of the present invention (nt-p65-TMD fusion protein) and the protein level of inflammatory cytokines.

Figure 2d shows the relative expression levels of IL-1β, IL-6, TNFα and IL-17A in the LPS-treated spleen after treatment with a composition for inhibiting inflammatory response after surgery according to an embodiment of the present invention. Figure 2d shows the results of the RT-PCR test by removing the spleen on the 2nd day after LPS administration. Specifically, the levels of inflammatory cytokines such as IL-1β, IL-6, TNFα and IL-17A were increased in the LPS-treated group, and in the case of the group administered with the composition of the present invention (nt-p65-TMD fusion protein), LPS When compared to the treatment group, it can be seen that the expression level of inflammatory cytokines is effectively reduced. As a result, the composition of the present invention (nt-p65-TMD fusion protein) has an effect of suppressing the expression level of the M1 phenotype of inflammatory cytokines.

Example 3: Evaluation of cognitive function through neurobehavioral test

In order to evaluate the efficacy of the composition according to an embodiment of the present invention on postoperative cognitive dysfunction, a model similar to abdominal surgery was used. A model similar to human abdominal major surgery was constructed using male BALB/c mice (6 to 8-week-old) after anesthesia with an inhalation anesthetic, ligating the gastrointestinal artery and rubbing the intestine with a finger. The composition of the present invention was administered in the postoperative recovery phase, and the following tests were performed.

Manual Avoidance Test-Assessment of Learning and Memory

In the passive avoidance test, the room with the same structure is divided into a room with lighting and a dark room without lighting (25 cm wide, 20 cm long, and 20 cm high), and a small connecting door is installed between the two rooms. This test is a method of measuring the working memory ability of a mouse, and is widely used for learning and measuring memory.

(i) Training: Turn on the light in the left room of the shuttle box and put the mouse's head facing the opposite side of the connecting door. After exploring the mouse for 10 seconds, open the connecting door to allow it to enter the dark compartment. The mouse instinctively moves to the dark right room while exploring the room. At this time, mice that do not enter the dark side within 120 seconds after the connection door is opened are excluded from the experiment. Once the mouse enters the dark side, the connection door closes. After that, an electric shock of 0.5 mA flows through the lattice-shaped floor for 3 seconds, and the mouse remembers it. After the connection door is opened, the time until the mouse enters the dark side is measured and recorded.

(ii) Test: The test is conducted to check the effect of surgery and anesthesia on memory 48 hours after training is over. Put the mouse in the round-trip box, and after the search time for 10 seconds, the connection door opens, and the latency time (retention time) of all four mouse feet entering the dark side is measured up to 300 seconds. The longer it takes to go to the dark, the better the learning and memory of passive avoidance is.

Referring to Figure 3a, the result of the passive avoidance test on the second day after surgery. The retention time was significantly reduced in the surgery-only group (POCD), but the treatment groups administered with 10 mg/kg of the composition of the present invention after surgery showed the same level of cognitive function improvement as in the normal group (contr o l). .

EPM (Elevated plus maze test)-working memory evaluation

Elevated plus maze (EPM) consists of two open counters, two closed counters and a central central platform. In this evaluation, the time the mice stayed in the open equilibrium for 5 minutes was measured. Of the four passages, two facing each other (30 cm long and 5 cm wide) are open, and the other two of the same size consist of a wall with a height of 20 cm. The central pulpit was 5 cm wide and 5 cm long, and a video camera was installed on the ceiling in the middle of the maze to record the animal's behavior.

At the beginning of the test, the mice were placed with their heads facing out on the open countertop of the maze and then allowed to freely navigate the maze. Behavior was observed for 5 minutes. Memory capacity was measured by using the SMART Video Tracking System and the SMART v2.5.21 program, respectively, for the time the mice stayed in the open and closed counterweights, and the total distance traveled.

On the first and second days of each mouse, the time required to enter the parallel band (Transfer latency) was measured regardless of any of the closed balance zones on both sides. What was learned here is defined as the time when the time required reduced on the second day was shown compared to the first day. For quantitative analysis, the learning index was calculated based on the relative difference between the time spent on the first and second days. A high learning index means that the hippocampus function in the brain is excellent.

In FIG. 3B, (a) is the time taken the day before surgery, (b) is the second day after surgery, and (c) shows the learning index. While the POCD group showed a low learning index, the nt-p65-TMD (10mg/kg) group treated with the composition of the present invention did not differ from the learning index of the control group. That is, it can be seen that the learning ability is improved due to the therapeutic effect of the composition of the present invention.

Cognitive test for new objects-cognitive memory evaluation

Place two identical objects on the inner center of the acrylic box (40 cm x 40 cm x 40 cm) at intervals of 18 cm and allow for 10 minutes to search. After 48 hours, one of the two was replaced with a new object of a different color and shape. The observation was made for a minute and the time the mouse stayed on the new object was measured.

It is performed for the evaluation of attention and concentration, and when presenting the same two objects to the mouse, curiosity is naturally induced and the mouse detects the two objects. At this time, the time for detecting the two objects is usually similar. However, if one of the two objects is replaced with a new object after 48 hours, the time to detect the new object is increased more because the mouse generally has more curiosity about the new object. In this example, the mouse was placed in an acrylic (40×40×40) cm box before surgery and was adapted to the environment in the box for 5 minutes. After 5 minutes, two objects were placed in the box and the reaction time for each object was observed and recorded for 3 minutes. 48 hours after the operation, one of the two objects was changed to a new object with a different color and shape, and then observed for 3 minutes to record the reaction time to the new object.

3C, there was no difference between groups in the search period for the same two objects before surgery, but the search time for a new object on the second day after surgery was the control group and the nt-p65-TMD (10mg/kg) group. While the search period for new objects increased, in the case of the POCD group, there was no difference between the search period for new objects and previous objects. This means a decrease in cognitive function after surgery, and indicates that cognitive memory capacity is improved as a therapeutic effect when the composition of the present invention is administered after surgery.

As a result of the above examples, the composition of the present invention (nt-p65-TMD fusion protein) effectively lowers the expression level or protein concentration of pro-inflammatory cytokines induced in the inflammatory response, as well as the expression level or protein of the anti-proinflammatory cytokine. By raising the level, it may have the effect of improving the inflammatory response that may occur after surgery. Furthermore, the composition of the present invention (nt-p65-TMD fusion protein) may have an effect of improving learning ability and memory. Accordingly, the composition of the present invention (nt-p65-TMD fusion protein) has an effect of preventing or treating cognitive dysfunction after surgery. At this time, improvement or suppression of the inflammatory response in the brain may lead to improvement of cognitive function. Accordingly, the composition for inhibiting inflammatory reaction after surgery according to an embodiment of the present invention may be provided as a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery according to another embodiment of the present invention.

Although the embodiments of the present invention have been described in more detail with reference to the accompanying drawings, the present invention is not necessarily limited to these embodiments, and various modifications may be made without departing from the spirit of the present invention. . Accordingly, the embodiments disclosed in the present invention are not intended to limit the technical idea of the present invention, but to explain the technical idea, and the scope of the technical idea of the present invention is not limited by these embodiments. Therefore, it should be understood that the embodiments described above are illustrative and non-limiting in all respects. The scope of protection of the present invention should be construed by the following claims, and all technical ideas within the scope equivalent thereto should be construed as being included in the scope of the present invention.

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION, YONSEI UNIVERSITY <120> COMPOSITION FOR TREATING OR PREVANTING DELIRIUM <130> 16PD5948KR-1 <160> 3 <170> KoPatentIn 3.0 <210> 1 <211> 187 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of p65 transcription modulation domain <400> 1 Met Asp Asp Leu Phe Pro Leu Ile Phe Pro Ser Glu Pro Ala Gln Ala 1 5 10 15 Ser Gly Pro Tyr Val Glu Ile Ile Glu Gln Pro Lys Gln Arg Gly Met 20 25 30 Arg Phe Arg Tyr Lys Cys Glu Gly Arg Ser Ala Gly Ser Ile Pro Gly 35 40 45 Glu Arg Ser Thr Asp Thr Thr Lys Thr His Pro Thr Ile Lys Ile Asn 50 55 60 Gly Tyr Thr Gly Pro Gly Thr Val Arg Ile Ser Leu Val Thr Lys Asp 65 70 75 80 Pro Pro His Arg Pro His Pro His Glu Leu Val Gly Lys Asp Cys Arg 85 90 95 Asp Gly Tyr Tyr Glu Ala Asp Leu Cys Pro Asp Arg Ser Ile His Ser 100 105 110 Phe Gln Asn Leu Gly Ile Gln Cys Val Lys Lys Arg Asp Leu Glu Gln 115 120 125 Ala Ile Ser Gln Arg Ile Gln Thr Asn Asn Asn Pro Phe His Val Pro 130 135 140 Ile Glu Glu Gln Arg Gly Asp Tyr Asp Leu Asn Ala Val Arg Leu Cys 145 150 155 160 Phe Gln Val Thr Val Arg Asp Pro Ala Gly Arg Pro Leu Leu Leu Thr 165 170 175 Pro Val Leu Ser His Pro Ile Phe Asp Asn Arg 180 185 <210> 2 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Hph-1 <400> 2 Tyr Ala Arg Val Arg Arg Arg Gly Pro Arg Arg 1 5 10 <210> 3 <211> 235 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of nt-p65-TMD <400> 3 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Ala Ser Gly Tyr Ala Arg Val Arg Arg Arg Gly 20 25 30 Pro Arg Arg Gly Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Glu Phe 35 40 45 Met Asp Asp Leu Phe Pro Leu Ile Phe Pro Ser Glu Pro Ala Gln Ala 50 55 60 Ser Gly Pro Tyr Val Glu Ile Ile Glu Gln Pro Lys Gln Arg Gly Met 65 70 75 80 Arg Phe Arg Tyr Lys Cys Glu Gly Arg Ser Ala Gly Ser Ile Pro Gly 85 90 95 Glu Arg Ser Thr Asp Thr Thr Lys Thr His Pro Thr Ile Lys Ile Asn 100 105 110 Gly Tyr Thr Gly Pro Gly Thr Val Arg Ile Ser Leu Val Thr Lys Asp 115 120 125 Pro Pro His Arg Pro His Pro His Glu Leu Val Gly Lys Asp Cys Arg 130 135 140 Asp Gly Tyr Tyr Glu Ala Asp Leu Cys Pro Asp Arg Ser Ile His Ser 145 150 155 160 Phe Gln Asn Leu Gly Ile Gln Cys Val Lys Lys Arg Asp Leu Glu Gln 165 170 175 Ala Ile Ser Gln Arg Ile Gln Thr Asn Asn Asn Pro Phe His Val Pro 180 185 190 Ile Glu Glu Gln Arg Gly Asp Tyr Asp Leu Asn Ala Val Arg Leu Cys 195 200 205 Phe Gln Val Thr Val Arg Asp Pro Ala Gly Arg Pro Leu Leu Leu Thr 210 215 220 Pro Val Leu Ser His Pro Ile Phe Asp Asn Arg 225 230 235

Claims (10)

delete delete delete As a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery,
A fusion protein comprising a transcriptional regulatory domain and a protein transport domain of NF-κB,
The transcriptional regulatory domain of NF-κB has the amino acid sequence of SEQ ID NO: 1,
The fusion protein has an amino acid sequence of SEQ ID NO: 3,
The cognitive dysfunction after the operation is a cognitive dysfunction due to brain inflammation induced after surgery, a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery. The method of claim 4, wherein
The fusion protein inhibits NF-κB transcription by competitive inhibition, thereby inhibiting braininflammation, preventing or treating cognitive dysfunction after surgery. The method of claim 4, wherein
The transcriptional regulatory domain of NF-κB is the transcriptional regulatory domain of RelA (p65), and the protein transport domain is Hph-1, wherein the pharmaceutical composition for preventing or treating cognitive dysfunction after surgery. The method of claim 4, wherein
The pharmaceutical composition comprises 0.0001% to 10% by weight of the fusion protein based on the total weight of the composition, a pharmaceutical composition for preventing or treating cognitive dysfunction after surgery. The method of claim 4, wherein
The composition is to be delivered to the brain, the pharmaceutical composition for preventing or treating cognitive dysfunction after surgery. The method of claim 4, wherein
The composition is a formulation for intraperitoneal administration, intranasal administration, intravenous administration, subcutaneous injection, intrathecal injection, inhalation administration, or oral administration, pharmaceutical composition for preventing or treating post-operative cognitive dysfunction. The method of claim 4, wherein
The composition is a dosage form for intraperitoneal administration or intranasal administration, pharmaceutical composition for preventing or treating cognitive dysfunction after surgery.

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