KR102107816B1 - Silk-derived peptide composition for improving cognitive function and memory - Google Patents

Abstract

The present invention relates to a silk-derived peptide composition for improving cognitive function and memory. According to the present invention, the silk-derived peptide composition for improving cognitive function and memory comprises: a first silk peptide composition which comprises bromelain and in which silk fibroin is hydrolyzed with a mixed enzyme containing any one selected from the group consisting of flavourzyme, protamex, neutrase, veron-W, sumizyme, zingibain, calpain, protease NP, validase, and mixtures thereof; and a second silk peptide composition in which silk fibroin is hydrolyzed with any one selected from the group consisting of delvorase, chymotrypsin, subtilisin, actinidin, lactoferrin, kokulase P, keratinase, and mixtures thereof.

Description

Silk-derived peptide composition for improving cognitive function and memory}

The present invention relates to a peptide composition derived from silk for improving cognitive function and memory. More specifically, it provides a peptide composition derived from silk for improving cognitive function and memory as well as having excellent initial cognitive function and memory, as well as excellent cognitive function and memory improvement effect.

Dementia is a complex clinical syndrome that manifests a marked impairment in human cognitive function, intellectual ability, emotional and behavioral changes, and leads to daily and social life due to cerebral cortical dysfunction such as memory, attention, language function, and spatiotemporal ability. It is a state of great difficulty. When defined as dementia, it refers to a case in which one or more of the other cognitive functions, including memory, is impaired.

The causes of dementia are degenerative brain diseases such as Alzheimer's Disease or Parkinson's Disease, brain hemorrhage, metabolic diseases of hepatic encephalopathy, Wilson's Disease, infectious diseases of neurosyphilis, AIDS and alcohol There are various etiologies such as drug addiction and brain trauma, and in some forms, diseases that can cause structural and functional abnormalities in the central nervous system can cause dementia. Among them, dementia caused by Alzheimer's disease is the most prevalent with 50 to 60%, followed by dementia caused by cerebrovascular disease.

Memory cognitive impairment is the first symptom experienced by people with Alzheimer's and is also the most common. In the early stages of Alzheimer's disease, patients suffer from a recent memory impairment that is unable to memorize recent conversations or details of work, due to damage to the hippocampal neurons and reduced ability to store recent memories. . However, during this period, remote long-term memory of events in the distant past is relatively well maintained. However, as the disease progresses, the cerebral cortex associated with the storage of long-term memory is damaged, and these memories of the past gradually become impaired.

On the other hand, if the region where the brain artery is blocked and the tissue dies due to a blood clot following arteriosclerosis occurs in an important area such as the hippocampus, it develops as dementia. Or subcortical ischemic vascular dementia (SIVD) (which accounts for 35 to 67% of total vascular dementia) due to poor blood supply (Roman et al., Subcortical ischaemic vascular dementia.Lancet Neurol 2002; 1) : 426-436). Damage in subcortical ischemic vascular dementia (SIVD) mainly occurs in white matter and various methods are used to make animal models resembling these diseases (Hainsworth & Markus, Do in vivo experimental models reflect human cerebral small vessel disease? A systematic review J Cerebral Blood Flow & Metabolism 2008; 28: 1877-1891). For vascular dementia, it has been found that oligodendrocytes, which form white matter, die by apoptosis in conditions where vascular dementia occurs, and studies are underway to suppress and treat them.

Currently, in the case of Alzheimer's disease treatment in Korea, foreign companies rely heavily on imports, accounting for 98% of the market share. Moreover, one company occupies 80% of the market, so it can be said that it is actually monopolized (Korea Institute of Science and Technology Information, dementia treatment. P.116, 2002). This shows that the growth of dementia treatment field is urgent even in terms of national competitiveness. Therefore, the development of an effective anti-antagonist and memory-improving agent with fewer side effects can significantly improve the quality of life of Alzheimer's disease and improve and treat early symptoms, and also help patients with forgetfulness and dementia who suffer from memory cognitive impairment. Can give.

KR 10-2019-0018897 A KR 10-2018-0094556 A

An object of the present invention is to provide a silk-derived peptide composition for improving cognitive function and memory, which is superior in cognitive function and memory improvement effect compared to the prior art using a hydrolase.

An object of the present invention is to provide a silk-derived peptide composition for improving cognitive function and memory, which can have a high improvement effect in the initial stage of administration with excellent activity in improving cognitive function and memory.

In addition, a combination of silk-derived peptides for improving cognitive function and memory with different mechanisms is provided to provide a silk-derived peptide composition having improved cognitive function and memory improvement effect and its initial reaction activity according to a complex effect.

In order to achieve the above object, the silk-derived peptide composition for improving cognitive function and memory according to an embodiment of the present invention includes bromelain, flavozyme, protamex, A group consisting of Neutrase, Veron-Double Oil, Sumyzyme, Zingibain, Calpain, Protease NP, Validase and mixtures thereof A first silk peptide composition in which silk fibroin is hydrolyzed with a mixed enzyme containing any one selected from, and delvorase, chymotrypsin, subtilisin, actinidin, lactoferrin (Lactoferrin), Kokasease P (Kokulase P), keratinase (Keratinase), and any one selected from the group consisting of mixtures of silk fibroin may be to include a hydrolyzed second silk peptide composition.

The silk-derived peptide composition for improving cognitive function and memory may be that the first silk peptide further comprises Umamizyme as a hydrolase.

In the silk-derived peptide composition for improving cognitive function and memory, the second silk peptide may further include cathepsin K as a hydrolase.

Food according to another embodiment of the present invention is to include a peptide composition derived from silk for improving the cognitive function and memory.

The drug for preventing or improving cognitive dysfunction according to another embodiment of the present invention is to include the peptide composition derived from silk for improving the cognitive function and memory.

The cognitive dysfunction is dementia, learning disability, real certification, forgetfulness, aphasia, agonism, delirium, AIDS-induced dementia, Vincewaner's disease, Lewy body dementia, frontotemporal dementia, mild cognitive impairment, multiple infarct dementia, pick disease, meaning It may be selected from the group consisting of dementia, Alzheimer's dementia or vascular dementia.

Hereinafter, the present invention will be described in more detail.

Silk-derived peptide composition for improving cognitive function and memory according to an embodiment of the present invention includes bromelain, flavozyme, protamex, Neutrase, and beron -Double oil (Veron), sumyzyme (Sumizyme), jingjingbain (zingibain), calpine (Calpain), protease NP (Protease NP), validase (Validase), and any one selected from the group consisting of mixture Silk silk fibroin hydrolyzed with a mixed enzyme, a first silk peptide composition and delvorase, chymotrypsin, subtilisin, actinidin, lactoferrin, and cocrase P (Kokulase P), keratinase (Keratinase), and any one selected from the group consisting of mixtures of silk fibroin may be to include a hydrolyzed second silk peptide composition.

Illustrating a method of preparing the first silk peptide composition, a dissolving step of dissolving the silk fibroin; A first proteolytic enzyme containing bromelain in the silk fibroin lysate and flavozyme, Protamex, Neutrase, Veron-Double Oil (Veron), Sumi A second proteolytic enzyme comprising any one selected from the group consisting of Sumzyme, Zingibain, Calpain, Protease NP, Validase and mixtures thereof. Hydrolysis by treatment with a containing enzyme; And an enzyme removal step of removing the proteolytic enzyme.

Silk (silk) refers to the fibers obtained from cocoons made by silkworms, and collectively refers to products made from silk yarns made from these. In a broad sense, it also refers to Yajamsa, such as Yakjamsa, and silk products from raw silk. The true silk, commonly called the dog, is also called the cultivated or domestic silk. Meanwhile, the cocoon in the present invention is defined as a concept including silk.

Silk fibers are composed of fibroin and sericin, and contain a small amount of brain, fat and inorganic salts. Silk consists of fibroin and sericin, which are fibrous proteins. It is composed of biopolymers in its natural state. Compounds with a molecular weight of more than 10,000 are usually fibers, membranes or tubers. Silk is a high-molecular protein, protein is made of amino acids, and amino acids are composed of simple alpha amino acids by combining amino groups (-NH ₂ ) and carboxyl groups (-COOH) on carbon.

In addition, a polymer having many chains of amino acids linked together in a rosary shape is called a polypeptide. Eventually, -NH ₂ and -COOH of the two amino acids react, and water (H ₂ O) escapes, and fiberization is achieved by arranging them in parallel.

The amino acids that make up fibroin have many glycine (side chain -H) and alanine (-CH ₃ ). These amino acids are characterized by a short side chain of the molecule. And the sum of serine (side chain-CH ₂ OH) and tyrosine (side chain-CH (OH) CH ₂ ) is more than 90%. The difference between the amino acid composition of sericin is that the content of acidic amino acids (aspartic acid, glutamic acid) and excitatory amino acids (lysine, alanine, histitin) is low. When these amino acids are combined in different linkage combinations, a polypeptide is formed. When hydrolyzed fibroin, it becomes an amino acid, and an alpha amino acid having a molecular weight of from 300 to 500,000 to 3,000 to 3,500 forms a single molecule.

The peptide referred to in the present invention refers to a peptide derived from silk, and has been processed by hydrolyzing silk fibroin derived from the silk to have a certain amino acid sequence in a size that can be absorbed into the body.

In addition, the peptide derived from silk referred to in the present invention can also be expressed as a silk protein and can be used in the same sense.

Preferably, the method for preparing a silk-derived peptide may further include a sericin removal step of removing the sericin by mixing and heat-treating the cocoon and purified water and dissolving the silk fibroin before the dissolving step of dissolving the silk fibroin. .

The sericin removal step refers to a step of removing the sericin surrounding the silk fibroin. Since sericin contains polar amino acids, it is common to remove it while performing heat treatment on purified water. Particularly, as the sericin removal step proceeds at a high temperature, the process efficiency of sericin removal increases, but there is a problem that the temperature cannot be increased indefinitely because the loss of fibroin occurs at a temperature exceeding 97 ° C.

In the method for producing a silk fibroin-derived peptide, the sericin removal step includes a thickener and an additive, and may be heat-treated at 30 to 70 ° C.

The sericin removal step may include a thickener and an additive, and heat treatment is performed at 30 to 70 ° C while heat treatment, and at the same temperature, the sericin removal process may be much faster than when the viscosity is not increased. In particular, when the viscosity is increased, there is an advantage that fibroin is not lost.

In the method for preparing the silk fibroin-derived peptide, the mixture of cocoon and purified water in the sericin removal step may have a viscosity of 1.2 to 120 cP.

According to the viscosity range, in the sericin removal process, not only can the sericin be quickly removed, but also has the advantage that there is little loss of fibroin.

Preferably, in the method for producing the silk fibroin-derived peptide, the sericin removal step is a mixture of cocoon and purified water having a viscosity of 1.3 to 40 cP, a first heat treatment step of heat treatment at 98 to 100 ° C for 2 to 10 minutes, and It may be to include a second heat treatment step of heat treatment to 30 to 40 ℃.

The first heat treatment step is the temperature at which fibroin is dissolved, but when the heat treatment is started by adjusting the viscosity of the cocoon and purified water to 1.3 to 40 cP, loss of fibroin may not occur.

Subsequently, if a relatively low secondary heat treatment step is performed, sericin may be rapidly removed.

In the first heat treatment step, when the viscosity of the mixture of cocoon and purified water is less than 1.3 cP, there is a problem that rapid loss of fibroin occurs. On the other hand, if the viscosity exceeds 40 cP, the process improvement effect for sericin removal may not appear despite the first heat treatment step.

Therefore, in the case of the primary heat treatment and the secondary heat treatment process while maintaining the viscosity range, the sericin can be removed more effectively, but loss of fibroin does not occur, thereby improving the efficiency of the sericin removal process.

When the additive is included, it is possible to minimize the loss of skin loin, as well as to increase the yield of fibroin hydrolysates, that is, silk fibroin-derived peptides, which exhibit the effect of improving memory and cognitive function in a later step.

Preferably, the additive contains xylase, and based on 100 parts by weight of xylase, sorbitol 10 to 100 parts by weight; Mannitol 1 to 10 parts by weight; Calactose may be included in 0.1 to 5 parts by weight.

In the case of the above range, the loss of fibroin in the sericin removal step is reduced, and the yield of the peptide derived from silk fibroin having a cognitive function improvement effect in the hydrolysis step is increased, and the efficiency of improving cognitive function and memory can be improved.

The dissolving step refers to preparing silk fibroin to be mixed with purified water, an organic solvent, and other liquids.

The first silk peptide contains bromelain in the silk fibroin lysate in the hydrolysis step, Flavozyme, Protamex, Neutrase, Veron-Double Oil (Veron) ), Sumyzyme, Zingibain, Calpain, Protease NP, Validase and any one selected from the group consisting of mixtures thereof. Refers to a hydrolyzate obtained by hydrolysis.

The second silk peptide is delvorase, chymotrypsin, subtilisin, actinidin, lactoferrin and lactferrin in the silk fibroin lysate in the hydrolysis step. (Kokulase P) refers to a hydrolyzate obtained by treating and hydrolyzing any one selected from the group consisting of keratinase and mixtures thereof.

Preferably, in the first silk peptide, the hydrolase may be any one selected from the group consisting of bromelain, jingzine, calpine, and mixtures thereof. According to the above range, it is possible to produce a silk fibroin peptide having a high effect of improving cognitive function and memory.

More preferably, in the first silk peptide, the hydrolytic enzyme contains bromelain, a mixed enzyme in which 10 to 50 parts by weight of Jingjibine and 5 to 30 parts by weight of calpine are mixed with respect to 100 parts by weight of bromelain. It may be used. According to the above range, the effect of improving cognitive function and memory can be excellent, and the effect of improving cognitive function and memory can be rapidly increased within a short period. Therefore, it is possible to produce a silk fibroin peptide capable of improving cognitive function and memory in a relatively short period of time.

Preferably in the second silk peptide, the hydrolase may be any one selected from the group consisting of Actinidin, Cocrase P, Keratinase and mixtures thereof. According to the above range, it is possible to produce a silk fibroin peptide having a high effect of improving cognitive function and memory.

More preferably, in the second silk peptide, the hydrolase contains actinidin, and 5 to 20 parts by weight of cocrase P and 5 to 20 parts by weight of keratinase are mixed with respect to 100 parts by weight of actinidine. It may be using a mixed enzyme. According to the above range, the effect of improving cognitive function and memory can be excellent, and the effect of improving cognitive function and memory can be rapidly increased within a short period. Therefore, it is possible to produce a silk fibroin peptide capable of improving cognitive function and memory in a relatively short period of time.

The enzyme removal step of removing the proteolytic enzyme refers to removing the enzyme or removing the activity of the enzyme after the hydrolysis step, and is defined to include all forms that can be used by those skilled in the art. .

In addition, the enzyme removal step may be to remove the activity of the enzyme by heat treatment.

In the first silk peptide, the proteolytic enzyme may further include Umamizyme. When the umizyme is included, the absorption rate of the hydrolyzate and the activity of the active ingredient for improving the cognitive function are increased, so that the cognitive function and memory improvement effect can be exhibited with a small amount in a shorter time.

Preferably, in the first silk peptide, the hydrolase contains bromelain, 10 to 50 parts by weight of Zynjibine, 5 to 30 parts by weight of calpine, 1 to 20 of umamizyme It may be using a mixed enzyme mixed with parts by weight. In the case of the above range, a silk-derived peptide having a significantly increased activity of an active ingredient for improving the absorption rate and cognitive function of a hydrolyzate can be prepared.

In the second silk peptide, the proteolytic enzyme may further include cathepsin K. When the umizyme is included, the absorption rate of the hydrolyzate and the activity of the active ingredient for improving the cognitive function are increased, so that the cognitive function and memory improvement effect can be exhibited with a small amount in a shorter time.

Preferably in the second silk peptide, the hydrolase contains actinin, 5 to 20 parts by weight of cocrase P, 5 to 20 parts by weight of keratinase, cathepsin with respect to 100 parts by weight of actinine K 1 to 5 parts by weight of the mixed enzyme may be used. In the case of the above range, a silk-derived peptide having a significantly increased activity of an active ingredient for improving the absorption rate and cognitive function of a hydrolyzate can be prepared.

The enzyme removal step may be to remove the activity of the enzyme by heat treatment.

Food according to another embodiment of the present invention is to include a peptide composition derived from silk for improving the cognitive function and memory.

The drug for preventing or improving cognitive dysfunction according to another embodiment of the present invention is to include the peptide composition derived from silk for improving the cognitive function and memory.

The cognitive dysfunction is dementia, learning disability, real certification, forgetfulness, aphasia, agonism, delirium, AIDS-induced dementia, Vincewaner's disease, Lewy body dementia, frontotemporal dementia, mild cognitive impairment, multiple infarct dementia, pick disease, meaning It may be selected from the group consisting of dementia, Alzheimer's dementia or vascular dementia.

The cognitive function and memory improvement referred to in the present invention are not only the meaning that the cognitive function and the memory slowdown are recovered or improved. It is defined as including the phenomenon of cognitive function and memory decline caused by brain neurological disease being prevented or improved.

A drug for preventing and improving brain neurological diseases according to another embodiment of the present invention may include silk-derived peptides prepared according to the method for preparing the silk-derived peptides.

The brain neurological disease referred to in the present invention is dementia, Huntington's disease, Parkinson's disease and neurodegenerative diseases such as amyotrophic lateral sclerosis, ischemic or reperfusion diseases such as ischemic stroke, depression, schizophrenia and post-traumatic stress disorder It is defined as including all mental disorders such as.

The silk-derived peptide composition for improving cognitive function and memory according to the present invention is significantly superior in cognitive function and memory improvement effect compared to the prior art using a hydrolytic enzyme.

In addition, the silk-derived peptide composition for improving cognitive function and memory according to the present invention has excellent activity in improving cognitive function and memory and can have a high improvement effect in the initial stage of administration.

In addition, the silk-derived peptide composition for improving cognitive function and memory according to the present invention is a combination of silk-derived peptide for improving cognitive function and memory having different mechanisms, and according to the complex action effect, the effect of improving cognitive function and memory and its initial reaction activity This can be greatly improved.

Figure 1 relates to the results of a cognitive memory evaluation test for silk peptides prepared according to an embodiment of the present invention.
Figure 2 is for the results of the effect experiment on the cerebral infarction site by ischemia of the silk peptide prepared according to an embodiment of the present invention.
Figure 3 relates to the results of the cognitive memory evaluation test for silk peptides prepared according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art to which the present invention pertains can easily practice. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein.

[Production Example 1: Preparation of peptide derived from first silk]

The dissolved silk fibroin was mixed with purified water, and the proteolytic enzymes T1 to T11 according to the composition as shown in [Table 1] were treated at 1w%, and hydrolysis was performed for 5 hours. After that, heat treatment was performed for 10 minutes to remove the activity of the enzyme, followed by freeze-drying, thereby preparing a powder as in Table 2.

T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 Flavozyme 100 - - - - - - - - - - Protamax - 100 - - - - - - - - - Bromelain - - 100 100 100 100 100 100 100 100 100 Jingji Vine - - - 5 10 30 50 60 30 30 30 Calpine - - - 5 10 20 30 40 20 20 20 Umamizyme - - - - - - - - 0.5 10 25

(Unit: parts by weight)

NF1 NF2 TF1 TF2 TF3 TF4 TF5 TF6 TF7 TF8 TF9 Processing enzyme T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

[ Experimental example  1: Article 1 silk  origin Peptide  Experiments on the effect of improving memory and cognitive function]

Experimental Example 1: Animal model experiment for improving cognitive function

A. Preparation of experimental animals

The experimental animals are male white paper (Sprague Dawley, 140-180 g, Korea Laboratory Animal Center), and they are put in 4 animals per breeding box for a week in a constant environment (room temperature 25 ± 1 ℃, relative humidity 60 ± 10%). Water was provided without restriction, and at this time, limited feeding was performed to maintain 80% of the weight. A total of 50 animals were used in this experiment. From 3 days before training, the test was conducted by adapting to the maze for 5 minutes.

B. delayed alternation test

There are a total of three passages (left, right, and center), and each passage entrance is prepared with a “ㅠ” shaped maze with partitions to allow the experimenter to control the entry and exit of animals.

In the first trial, the starting box was adapted for 1 minute. The first trial was a 'forced selection trial', where one passage was blocked with a partition and the back was blocked with a partition when an animal entered the other passage. Animals were considered to have entered when the limbs of the animals had passed through the entrance to the passage. Animals were fed for 60 seconds in the passage.

From the next trial, the 'free selection trial' allowed the animals to freely select the direction of the passage (left or right) starting from the starting box. Animals were compensated for prey only when they entered the passage from the previous trial and into the other.

The interval between trials was 5 seconds and 20 seconds with a delay, and the cut-off time for each trial was 60 seconds. The number of times the animal entered the passage through which the reinforcement was given and the time taken to enter the passage were measured. The same procedure was performed for 9 days of 10 trials a day to evaluate.

C. Method of administration

Silk peptides NF 1, 2 and TF 1 to TF 9 prepared according to the preparation method of the present invention were prepared as a solution of 100 mg / ml and administered orally. All samples were dissolved in distilled water and administered orally.

D. Results of cognitive memory evaluation

The results of the T-maze experiment for the groups administered with silk peptides NF 1, 2 and TF 1 to TF 9 are shown in [Fig. 1] below. [Fig. 1] shows that the experiment was conducted by configuring the trial interval as 5 seconds, and the performance gradually improved during the training period of 9 days.

The present inventors were able to derive results having a similar tendency as in the case of using a mixed enzyme centered on papain in other experiments conducted at the same time. In this case, it can be confirmed that the improvement effect on cognitive function is apparent. In addition, it was found that in the case of the above production example, the early improvement effect was also excellent.

In particular, it can be seen that TF8 exhibits an excellent effect of improving initial cognitive function compared to other preparation examples. It can be understood that the peptide having a specific sequence in the silk fibroin hydrolyzed by the mixed hydrolase is mixed in a certain range and accelerates the effect of improving cognitive function, thereby exhibiting a rapid effect at the initial administration.

As a result of this analysis, it is considered that the above acceleration effect may occur when peptides containing sequences represented by the following [General Formula 1] and [General Formula 2] are simultaneously included.

[Formula 1]

Tyr-Gly-X ₁ -X ₂

(In the general formula, X ₁ is Ala, Tyr, Val or Gln, and X ₂ is Ala, Tyr, Ile, Val or Gly.)

[Formula 2]

Ile-Gly-Z ₁ -Z ₂

(In the above general formula, Z ₁ is Ala, Tyr, Val or Gln, and Z ₂ is Ala, Tyr, Val or Gly.)

Therefore, in the case of the above range, it is possible to provide a peptide derived from silk having an effect of improving cognitive function and memory, and in particular, it is possible to accelerate an initial synergistic effect on improvement of cognitive function and memory, and the effect of memory and cognitive function is further improved. Silk peptides can be prepared.

Experimental Example 2: Local ischemic induction animal model

Evaluation of the effect on the area of cerebral infarction due to ischemia using a local ischemia-induced animal model to confirm the degree of improvement in memory and cognitive function for silk peptides prepared using the hydrolytic enzymes of NF1, NF2, TF5 and TF8 The experiment was conducted.

A. Drug treatment and construction of a local ischemic induction animal model

A local ischemic induction animal model was constructed using a male white paper (Sprague-Dawley rat) weighing 200-250 g. The drug was administered orally with 1 g / kg of NF1, NF2, TF5 and TF8 1 hour before induction of ischemia or 1 hour after induction of ischemia (oral administration group, n = 6), and ischemia control group (n = 6). BG101 was administered in the same dose as the silver drug treatment group. Experimental animals were anesthetized by intramuscular injection of ketamine (30 to 40 mg / kg), followed by skin incision in the neck in the supine position, common carotid artery, external carotid artery, and internal carotid. artery) and separated from surrounding tissue. First, electrocauterize the upper thyroid gland and larynx and arteries, which are branches of the external carotid artery, electrocauterize the pterygopalatine artery, which is the branch of the internal carotid artery, and cut the external carotid artery to remove the 4-0 nylon thread (ETHICON, INC), etc. After inserting into the internal carotid artery through the carotid artery, the nylon cerebral artery was occluded by placing the nylon thread 16 to 18 mm in the carotid artery branch.

B. Experiment of reduction effect of cerebral infarction due to ischemia

After 6 hours of induction of ischemia, the animals were isolated and the brain was isolated. The extracted brain was sectioned at 2 mm intervals from the anterior pole, reacted with 2% triphenyl tetrazolium chloride (TTC, Sigma) solution at 37 ° C for 30 minutes, and then 4% paraformaldehyde ( Sigma) solution and fixed. After taking the stained tissue sections, the area of the area that turned white due to the occurrence of cerebral infarction was measured using the MCID image processing system (Imaging Research Inc.) and dyed red, and the area of the entire brain section was measured. After calculating the ratio of the area of contrast infarction, the average value was calculated. In addition, the ratio of the volume occupied by the cerebral infarction area in the total volume in each brain section was measured. The results are shown in [Figure 2].

Referring to FIG. 2, it can be confirmed that, in the case of the silk glass fibroin provided by the present invention, the cerebral infarction can be reduced.

[Production Example 2: Preparation of peptide derived from second silk]

In the same manner as in Preparation Example 1, the hydrolysis of silk fibroin was performed by treating the cocktail proteolytic enzymes S1 to S11 of Table 3 alone. After that, heat treatment was performed for 10 minutes to remove the activity of the enzyme, followed by freeze-drying to prepare a powdery form as shown in [Table 4].

S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 Flavozyme 100 - - - - - - - - - - Protamax - 100 - - - - - - - - - Actinine - - 100 100 100 100 100 100 100 100 100 Cokrase P - - - One 5 10 20 30 10 10 10 Keratinase - - - One 5 10 20 30 10 10 10 Card Depsin K - - - - - - - - 0.5 3 6

(Unit: parts by weight)

NF1 NF2 TP1 TP2 TP3 TP4 TP5 TP6 TP7 TP8 TP9 Processing enzyme S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11

[ Experimental example  2: Article 2 silk  origin Peptide  Experiments on the effect of improving memory and cognitive function]

In the same manner as in Experimental Example 1, the results of the T-maze experiment for the group administered with the silk peptides NF 1, 2 and TP 1 to TP 9 are shown in [Fig. 3] below. The present inventors were able to derive similar tendency results when using a mixed enzyme centered on papain in other experiments conducted simultaneously. In this case, it can be confirmed that the improvement effect on cognitive function is apparent. In addition, it was found that in the case of the above production example, the early improvement effect was also excellent.

In particular, it can be seen that TP8 exhibits an excellent effect of improving initial cognitive function compared to other preparation examples. It can be understood that the peptide having a specific sequence in the silk fibroin hydrolyzed by the mixed hydrolase is mixed in a certain range and accelerates the effect of improving cognitive function, thereby exhibiting a rapid effect at the initial administration.

As a result of this analysis, a peptide comprising an amino acid sequence represented by the following [General Formula 1a] or [General Formula 1b]; It is believed that the above-described acceleration effect may occur when peptides containing an amino acid sequence represented by the following [General Formula 2] are simultaneously included.

[Formula 1a]

a ₁ -Gly-Gly-a ₁

(In the above general formula, a ₁ is Ala, Tyr, or Val)

[General Formula 1b]

a ₁ -Gly-Gly-a ₁ -Gly-a ₂

(In the above general formula, a ₁ is Ala, Tyr, or Val, and a ₂ is Ala, Tyr, Ile or Val.)

 [Formula 2]

Ile-Gly-Z ₁ -Z ₂

(In the above general formula, Z ₁ is Ala, Tyr, Val or Gln, and Z ₂ is Ala, Tyr, Val or Gly.)

Therefore, in the case of the above range, it is possible to provide a peptide derived from silk having an effect of improving cognitive function and memory, and in particular, it is possible to accelerate an initial synergistic effect on improvement of cognitive function and memory, and the effect of memory and cognitive function is further improved. Silk peptides can be prepared.

Accordingly, the first silk peptide composition and the second silk peptide composition may have cognitive function and memory improvement effect, respectively, and have characteristics of excellent early improvement activity.

Furthermore, it is considered that there is a difference in sequence composition in the analyzed peptide composition, and a mechanism in which each activity is derived. Therefore, when the mixture of the first silk peptide composition and the second silk peptide composition is used, it is expected that the effect of improving cognitive function and memory and improving the initial activity may be higher by different mechanisms.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also provided. It belongs to the scope of rights.

Claims (6)

Silk fibroin is the first mixed enzyme containing bromelain, mixed with 10 to 50 parts by weight of zingibain and 5 to 30 parts by weight of calpain, based on 100 parts by weight of bromelain. Disassembled first silk peptide composition and
As a second mixed enzyme containing Actinidin, 5 to 20 parts by weight of Kokase P (Kokulase P), and 5 to 20 parts by weight of Keratinase (Keratinase) with respect to 100 parts by weight of Actinidin A silk fibroin hydrolyzed second silk peptide composition comprising
A peptide composition derived from silk for improving cognitive function and memory. According to claim 1,
The first mixed enzyme further comprises Umamizyme
A peptide composition derived from silk for improving cognitive function and memory. According to claim 2,
The second mixed enzyme further comprises cathepsin K (Cathepsin K)
A peptide composition derived from silk for improving cognitive function and memory. A food product comprising the peptide composition according to claim 1 or 2. A drug for preventing or improving cognitive dysfunction comprising the peptide according to claim 1 or 2. The method of claim 5,
The cognitive dysfunction is dementia, learning disability, real authentication, forgetfulness, aphasia, agonism, delirium, AIDS-induced dementia, Vincewaner's disease, Lewy body dementia, frontotemporal dementia, mild cognitive impairment, multiple infarct dementia, pick disease, meaning Is selected from the group consisting of dementia, Alzheimer's dementia or vascular dementia
medicine.

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