Composition containing pentapeptide as an active ingredient

Abstract

To suppress body odor by suppressing excessive sweating, and to develop a new therapeutic agent for pain.SOLUTION: The present invention relates to the use of a composition comprising a pentapeptide as an active ingredient to suppress excessive sweating, to suppress body odor or to prevent or treat pain.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to use for suppressing excessive sweating, use for suppressing body odor, or use for preventing or treating pain of a composition containing a pentapeptide as an active ingredient.

Background Art

[0002] Hyperhidrosis means a skin disorder characterized by sweating and sweat secretion that are excessively and abnormally increased more than necessary for body temperature regulation. Hyperhidrosis is caused by genetic factors or physical and emotional stress, and may appear globally or locally in specific parts of the body. When excessive sweating appears locally on the palms, soles, face, armpits, scalp, etc., this is called primary or local hyperhidrosis. Generalized or secondary hyperhidrosis usually affects the whole body.

[0003] Currently, methods for treating hyperhidrosis include using antiperspirants, iontophoresis to temporarily prevent sweat glands, Botox therapy (WO2010-078242) to temporarily block the nerves that stimulate sweating, and surgical methods such as endoscopic thoracic sympathectomy and removing sweat glands. Among these, in the case of the method of temporarily blocking nerves using botulinum toxin, since its paralytic effect is reversible for an average of 6 months, repeated injections of botulinum toxin are required for such treatment. Since botulinum toxin has a size that can be recognized by the patient's immune system, an immune reaction to the drug may be induced. Therefore, there is a need to develop a molecule that has a simpler and more stable molecular structure that does not induce an immune reaction, has high sweat-reducing activity, has long persistence, and enables non-invasive treatment with few side effects.

[0004] On the other hand, body odor is caused by the decomposition of sweat secreted from sweat glands by microorganisms and bacteria. The fats, proteins, sebum, and pollutants contained in sweat secreted from eccrine sweat glands, which are distributed throughout the body, and apocrine sweat glands, which are distributed in areas with a lot of body hair such as the armpits, are decomposed by microorganisms and bacteria, resulting in unpleasant body odors such as underarm odor, foot odor, and scalp odor. Generally, methods such as deodorizing, antibacterial, and masking are used to control body odor, but using the wrong fragrance to hide body odor can have the opposite effect, making the odor worse. In addition, when deodorizing components are used on the skin, they only provide a temporary effect after treatment until the next wash, and there is the inconvenience of having to re-treat after washing.

[0005] Therefore, the inventors made diligent efforts to develop a material that can suppress body odor by suppressing the production of excessive sweat, which can be considered the root cause of body odor. As a result, they confirmed that the pentapeptide newly synthesized by the inventors has a stable molecular structure, penetrates to the skin tissue through the skin, and exhibits high sweat-reducing activity, and can be usefully used for the treatment of hyperhidrosis and the control of body odor, thus completing the present invention.

[0006] On the other hand, pain can arise from a variety of causes, and if it persists for a long time or the stimulus is too severe, it can interfere with daily life and cause anxiety and fear. Generally, pain is called headache, chest pain, abdominal pain, lower back pain, etc., depending on the location of its occurrence, and is classified into somatic pain, visceral pain, neuropathic pain, psychogenic pain, etc., depending on the mechanism of occurrence, and is divided into acute pain and chronic pain based on the concept of time. Chronic pain, which has clinical significance, is generally characterized by pain that persists for more than six months despite the absence of a clear cause for the pain. The majority of chronic pain manifests as neuropathic pain characterized by spontaneous pain, allodynia, and hyperalgesia, and is characterized by severe pain that can be triggered by slight temperature changes or light contact with clothing, causing significant interference with the patient's daily life, and not responding well to various existing treatments.

[0007] Currently, pain is treated with invasive therapies such as nerve block and many types of drug therapies (opioid drugs, drugs that modulate the activity of nerve ion pathways, anticonvulsants, etc.). However, these drugs only show their effects when administered in high doses, and even if they are effective initially, tolerance often develops with long-term use, rendering them ineffective. In addition, side effects such as gastrointestinal bleeding, gastrointestinal disorders, and liver damage may occur.

[0008] Therefore, a new treatment method using botulinum toxin A has been introduced as a way to eliminate the side effects of existing treatments. Ranoux et al. reported a reduction in pain and allodynia after intradermal injection of botulinum toxin in patients with postherpetic pain and postoperative neuropathic pain who also had mechanical allodynia (Ranoux et al., Annals of Neurology 64(3):274~283(2008)), and Yuan et al. reported a reduction in pain and improvement in sleep disturbances in patients with diabetic neuropathy (Yuan et al., Neurology 72:1473~1478(2009)). However, recently there have been increasing reports of the same effect being observed when administering stomach medication, so more research results are needed.

[0009] On the other hand, botulinum toxin has a reversible effect that lasts an average of six months, requiring repeated injections to treat pain. Furthermore, it is large enough to be recognized by the patient's immune system, which can trigger an immune response to the drug. Therefore, there is a need to develop a molecule that does not trigger an immune response, has a simpler and more stable molecular structure, possesses high pain-relieving activity, has long-lasting effects, and offers non-invasive treatment with fewer side effects.

[0010] Therefore, the inventors, through diligent efforts to develop a new pain treatment agent, confirmed that the newly synthesized pentapeptide has a stable molecular structure, penetrates to the skin tissue via the skin, and exhibits high pain-suppressing activity, thus confirming that it can be usefully used in pain treatment, and thus completed the present invention. [Overview of the Initiative] [Problems that the invention aims to solve]

[0011] One object of the present invention is to provide a pharmaceutical composition for preventing or treating hyperhidrosis.

[0012] Another object of the present invention is to provide a method for preventing or treating hyperhidrosis.

[0013] Another object of the present invention is to provide peptides for use in the prevention or treatment of hyperhidrosis.

[0014] Another object of the present invention is to provide a composition for preventing or alleviating hyperhidrosis or for suppressing body odor.

[0015] Another object of the present invention is to provide a method for suppressing body odor.

[0016] Another object of the present invention is to provide peptides for use in suppressing body odor.

[0017] Another object of the present invention is to provide a composition for relieving pain.

[0018] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating pain.

[0019] Another object of the present invention is to provide a method for preventing or treating pain.

[0020] Another object of the present invention is to provide peptides for use in the prevention or treatment of pain.

[0021] Another object of the present invention is to provide a quasi-drug composition for relieving or improving pain.

[0022] Another object of the present invention is to provide a cosmetic composition for relieving or improving pain. [Means for solving the problem]

[0023] Therefore, one aspect of the present invention is to provide a pharmaceutical composition for the prevention or treatment of hyperhidrosis, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0024] Another aspect of the present invention provides a method for preventing or treating hyperhidrosis, which includes administering to a subject a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1.

[0025] Another aspect of the present invention provides a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 for use in preventing or treating hyperhidrosis.

[0026] Another aspect of the present invention provides a composition for preventing or alleviating hyperhidrosis or suppressing body odor, which contains as an active ingredient a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1.

[0027] Another aspect of the present invention provides a method for suppressing body odor, which includes administering to a subject a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1.

[0028] Another aspect of the present invention provides a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 for use in suppressing body odor.

[0029] Another aspect of the present invention provides a composition for alleviating pain, which contains as an active ingredient a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1.

[0030] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating pain, which contains as an active ingredient a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1.

[0031] Another aspect of the present invention provides a method for preventing or treating pain, which includes administering to a subject a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1. ?[[ID=?]]

[0032] [[ID=?]] Another aspect of the present invention provides a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 for use in preventing or treating pain. <0000?11> It seems there is some issue with the tags in the original text (e.g., etc. might be incorrect in the original). But I've translated it as accurately as possible based on the provided rules.Another aspect of the present invention is to provide a quasi-drug composition for the prevention or improvement of pain, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0034] Another aspect of the present invention is to provide a pain-relieving cosmetic composition containing a pentapeptide, which is composed of the amino acid sequence of SEQ ID NO: 1, as an active ingredient. [Effects of the Invention]

[0035] The pentapeptide composed of the amino acid sequence of Sequence ID No. 1 of the present invention is absorbed transdermally (through the skin) to the muscle layer of skin tissue, and can reduce sweat production by suppressing the abnormal release of neurotransmitters from nerve endings. Therefore, it can be used to prevent, alleviate, improve, or treat the symptoms of hyperhidrosis simply by applying it topically.

[0036] Furthermore, the pentapeptide of the present invention can suppress excessive sweat production, thereby blocking the cause of body odor in advance and providing a semi-permanent body odor suppression effect.

[0037] The pentapeptide composed of the amino acid sequence of Sequence ID No. 1 of the present invention can suppress the expression of infectious cytokines in muscle cells and inhibit the release of pain transmission factors from nerve endings, thereby suppressing the onset and transmission of pain. Therefore, it can be used for the purpose of alleviating, improving, preventing, or treating pain.

[0038] Furthermore, since the pentapeptide of the present invention is absorbed transdermally (through the skin) to the muscle layer of skin tissue, it can be used as a raw material for pharmaceuticals, quasi-drugs, and cosmetics applied to the skin.

[0039] However, the effects of the present invention are not limited to those mentioned above, and other effects not mentioned can be clearly understood by those skilled in the art from the following description. [Brief explanation of the drawing]

[0040] [Figure 1a]This figure shows the results of the thermal stability evaluation of KFLIK peptide: it shows the results of confirming the high-temperature stability of the peptide during long-term storage at 45°C. [Figure 1b] This figure shows the results of the thermal stability evaluation of the KFLIK peptide: it shows the results of confirming the high-temperature stability of the peptide at the maximum heating temperature (121°C). [Figure 2a] This figure shows the inhibitory effect of KFLIK peptide treatment on SNARE complex formation in nerve cells: the results of confirming the presence or absence of SNARE complex formation by Western blotting are shown. [Figure 2b] This figure shows the inhibitory effect of KFLIK peptide treatment on SNARE complex formation in nerve cells: it shows the results of confirming the degree of degradation of syntaxin 1a, a component of the SNARE complex. [Figure 3] This figure shows the inhibitory effect of KFLIK peptide treatment on neurotransmitter release in nerve cells. [Figure 4] This figure shows the relative sweat reduction effect of the KFLIK peptide treatment of the present invention on the palm area of ​​a patient with hyperhidrosis. A is an image showing the results of an iodine-starch experiment, and B shows the sweat reduction effect as an intensity analysis result using ImageJ software. [Figure 5] This figure shows the relative sweat reduction effect of the KFLIK peptide treatment of the present invention on the soles of the feet of patients with hyperhidrosis. Figure A is an image showing the results of an iodine-starch experiment, and Figure B shows the sweat reduction effect as a result of intensity analysis using ImageJ software. [Figure 6] This figure shows the relative sweat reduction effect of the KFLIK peptide treatment of the present invention in the armpit area of ​​hyperhidrosis patients. A is an image showing the results of an iodine-starch experiment, and B shows the sweat reduction effect as a result of intensity analysis using ImageJ software. [Figure 7]These are fluorescence microscope images confirming whether the KFLIK peptide penetrates into nerve cells after being attached to the fluorescent substance rhodamine: A shows the coexistence of the blue-stained nerve cell nucleus and the red rhodamine-pentapeptide; the blue in B shows the nerve cell nucleus stained with DAPI; and the red in C shows the rhodamine-peptide. [Figure 8] These are fluorescence micrographs taken from the backs of experimental animals after applying rhodamine-KFLIK peptide, to confirm whether the KFLIK peptide penetrated into the muscle layer: A (blue) shows cell nuclei stained with DAPI, B (green) shows the TrkB nerve fiber marker, C (red) shows the rhodamine-pentapeptide, and D (yellow) shows that the pentapeptide of the present invention penetrated into the nerve cells of the muscle layer. [Figure 9] This figure illustrates the body odor-suppressing effect of the KFLIK peptide of the present invention, and the inhibitory effect on Corynebacterium proliferation is shown by absorbance measurement results using a spectrophotometer. P / C represents the positive control group, which is 1% triclosan. [Figure 10a] This figure shows the effect of KFLIK peptide treatment on suppressing the expression of infectious cytokines in muscle cells: The results show the mRNA expression levels of IL-1α, TNF-α, IL-1β, and COX-2 genes confirmed by RT-PCR. [Figure 10b] This figure shows the effect of KFLIK peptide treatment on suppressing the expression of infectious cytokines in muscle cells: it shows the results of confirming the expression levels of COX-2, IL-1β, and TNF-α proteins via Western blotting. [Figure 11] This figure shows the effect of KFLIK peptide treatment in nerve cells on suppressing the release of substance P, a pain transmission factor. [Figure 12a]This figure shows the muscle pain relief effect of KFLIK peptide treatment according to the present invention, based on human efficacy evaluation: The average pain intensity was calculated by measuring the Numeric Rating Scale (NRS). [Figure 12b] This figure shows the muscle pain relief effect of KFLIK peptide treatment according to the present invention, based on human efficacy evaluation: It shows the degree of pain reduction by KFLIK peptide treatment. [Modes for carrying out the invention]

[0041] The present invention will be described in detail below.

[0042] One aspect of the present invention is to provide a pharmaceutical composition for the prevention or treatment of hyperhidrosis, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0043] In this specification, the term "peptide" means a linear or cyclic molecule formed by the bonding of amino acid residues to each other via peptide bonds. The preparation of such peptides may be achieved by conventional biological or chemical synthesis methods known in the art, for example, by solid-phase synthesis techniques.

[0044] In this specification, the term "pentapeptide" means a linear molecule consisting of five amino acid residues, and the pentapeptide of the present invention specifically means a linear peptide molecule composed of the amino acid sequence (KFLIK) of SEQ ID NO: 1.

[0045] The aforementioned "peptides" and "pentapeptides" may be amino acid variants or fragments having different sequences due to deletions, insertions, substitutions, or combinations thereof of amino acid residues, to the extent that they do not affect the function. Amino acid exchanges that do not alter the overall activity of the peptides are known in the art. In some cases, they may be modified by phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc. Accordingly, the present invention includes peptides and variants thereof, or active fragments thereof, having an amino acid sequence substantially identical to a pentapeptide composed of the amino acid sequence of KFLIK. The substantially identical proteins mean amino acid sequences having 75% or more, preferably 80% or more, for example, 85% or more, 90% or more, 95% or more, 98% or more, or 99% or more sequence homology with the amino acid sequence of KFLIK, but are not limited thereto; any protein having 75% or more amino acid sequence homology and the same activity is included in the scope of the present invention. Furthermore, the peptides of the present invention may additionally include targeted sequences, tags, labeled residues, or amino acid sequences manufactured for specific purposes to increase half-life or peptide stability.

[0046] Furthermore, protecting groups may be attached to the N-terminus or C-terminus of the peptide of the present invention to obtain better chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., broad biological activity spectrum), and reduced antigenicity. For example, the protecting group may be an acetyl group, a fluorenyl methoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, or polyethylene glycol (PEG), but may include any component that can modify the peptide, particularly enhance its stability. The term "stability" is used to include not only in vivo stability that protects the peptide of the present invention from attack by endogenous protein-cutting enzymes, but also storage stability (e.g., room temperature storage stability).

[0047] The pentapeptide composed of the amino acid sequence of KFLIK according to the present invention can prevent or treat hyperhidrosis by inhibiting the formation of the SNARE complex and suppressing the excessive release of neurotransmitters from nerve endings.

[0048] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in the present invention exhibits an effect of suppressing sweat production in skin areas where hyperhidrosis occurs, and can prevent or alleviate hyperhidrosis.

[0049] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in the present invention penetrates to the muscle layer of skin tissue via transdermal absorption, so it can improve or alleviate the symptoms of hyperhidrosis simply by applying it to the affected area.

[0050] In this invention, "hyperhidrosis" means a disease condition characterized by excessive and uncontrolled sweating beyond what is necessary to cool the body, and refers to a disease or symptom in which excessive sweating occurs due to hyperfunction of the sweat glands and disruption of their cholinergic stimulation. In this invention, hyperhidrosis includes both localized hyperhidrosis and generalized hyperhidrosis, and may occur in any one skin area selected from the group consisting of, for example, the face, forehead, neck, hands, feet, and armpits, but is not limited to these.

[0051] In specific examples, when a composition containing the pentapeptide of the present invention was applied to one palm, sole of the foot, and armpit of a patient with hyperhidrosis, it was confirmed that sweat production was reduced by 45% to 90% compared to a control area on the opposite side where the composition was not applied.

[0052] In the present invention, the term "prevention" means all actions that suppress or delay pain by administering a composition containing the pentapeptide.

[0053] In the present invention, the term "treatment" means all actions in which pain is "relieved," "improved," or "modified" by the administration of the composition containing the pentapeptide.

[0054] The pharmaceutical compositions for the prevention or treatment of hyperhidrosis according to the present invention can be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, topical preparations, suppositories, and sterile injection solutions, respectively, by conventional methods, and may contain appropriate carriers, excipients, or diluents commonly used in the manufacture of pharmaceutical compositions for dosage formulation.

[0055] The pharmaceutically acceptable carriers may include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, and others commonly used in formulation.

[0056] The pharmaceutical composition may further contain, in addition to the above-mentioned components, lubricants, humectants, sweeteners, flavoring agents, emulsifiers, suspension agents, preservatives, and the like.

[0057] The pharmaceutical composition may be administered orally or parenterally (e.g., intramuscularly, intravenously, intraperitoneally, subcutaneously, intradermally, or topically) by the method of the purpose, and the dosage will vary depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the time, but may be appropriately selected by those skilled in the art.

[0058] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective dose may be determined by factors including the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the elimination ratio, the duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other obesity therapeutic agents, and may be administered simultaneously, separately, or sequentially with conventional therapeutic agents, and may be administered as a single or multiple dose. It is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, taking all of the above factors into consideration, and this can be easily determined by those skilled in the art.

[0059] The effective amount of the pharmaceutical composition may vary depending on the patient's age, sex, condition, weight, absorption rate of the active ingredient into the body, inactivation rate, excretion rate, type of disease, and concomitant drugs. It may also be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc. For example, the pharmaceutical composition may be administered at a dose of approximately 0.0001 μg to 500 mg, preferably 0.01 μg to 100 mg, per kg of the patient's body weight per day.

[0060] In other aspects, the present invention provides a composition for preventing or alleviating hyperhidrosis or suppressing body odor, comprising a pentapeptide composed of the amino acid sequence of KFLIK as an active ingredient.

[0061] The pentapeptide composed of the amino acid sequence of KFLIK in this invention can prevent or treat hyperhidrosis by inhibiting the formation of SNARE complexes and suppressing the excessive release of neurotransmitters from nerve endings, and can also suppress body odor caused by excessive sweating.

[0062] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in the present invention exhibits the effect of suppressing sweat production in skin areas where hyperhidrosis occurs, thereby preventing or alleviating hyperhidrosis and suppressing body odor caused by excessive sweat production.

[0063] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in the present invention can suppress the decomposition of substances contained in sweat and the generation of body odor by inhibiting the growth of body odor-inducing bacterial strains.

[0064] The specific details of the peptide, pentapeptide, and hyperhidrosis are as described above.

[0065] In this specification, "body odor" means the malodor of decomposition products generated when sweat gland secretions, i.e., fats, proteins, sebum and / or waste products excreted with sweat, are broken down by bacteria, and specifically, it may be a malodor that originates from body parts where a lot of sweat is produced, such as the armpits, feet, hands, forehead, neck, face, or scalp.

[0066] In this specification, "body odor suppression" means preventing malodors caused by products of decomposition of sweat gland secretions by microorganisms or suppressing the production of sweat that causes body odor. Specifically, "body odor suppression" may mean preventing malodors in the armpits, feet, hands, forehead, neck, face, or scalp, or suppressing sweat production in those areas, and more specifically, it may mean preventing bromhidrosis (bromhidrosis).

[0067] In another aspect, the present invention provides a cosmetic composition for preventing or alleviating hyperhidrosis or suppressing body odor, which contains a pentapeptide composed of the amino acid sequence of KFLIK as an active ingredient.

[0068] The cosmetic composition may be manufactured in any dosage form commonly produced in the art, such as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleanser, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited to these.

[0069] The aforementioned cosmetic composition may be manufactured in a variety of forms, such as softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, powder, deodorant, hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, gel, or other solutions, sol-gels, emulsions, oils, waxes, and aerosols, but is not limited to these forms.

[0070] The cosmetic composition of the present invention may contain other additives such as excipients and carriers in addition to the pentapeptide composed of the amino acid sequence of KFLIK, and it is possible to apply and incorporate ordinary ingredients commonly used in general skin cosmetics in the required amounts.

[0071] If the dosage form of the cosmetic composition is a paste, cream, or gel, animal oils, vegetable oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as the carrier component.

[0072] When the dosage form of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier component, and in particular when it is a spray, propellants such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether may be additionally included, but are not limited to these.

[0073] If the dosage form of the cosmetic composition is a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as the carrier component. For example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan may be used.

[0074] If the dosage form of the cosmetic composition is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol, a suspension agent such as ethoxylated isostearyl alcohol, polyoxyethyl sorbitol ester, or polyoxyethylene sorbitan ester, or microcrystalline cellulose, aluminum methhydroxyl, bentonite, agar, or tragacanth.

[0075] If the dosage form of the cosmetic composition is a surfactant-containing cleanser, then the carrier component may be an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinate monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0076] If the dosage form of the cosmetic composition is a hair shampoo, the trolox-peptide conjugate of the present invention may be mixed with base components for forming the shampoo, such as a thickener, surfactant, viscosity modifier, humectant, pH adjuster, preservative, and essential oil. CDE may be used as the thickener, LES, an anionic surfactant, and cocobetaine, an amphoteric surfactant, may be used as the surfactant, polyquartz may be used as the viscosity modifier, glycerin may be used as the humectant, and citric acid and sodium hydroxide may be used as the pH adjuster. Grapefruit extract may be used as the preservative, and essential oils such as cedarwood, peppermint, and rosemary, as well as silk amino acids, pentaol, or vitamin E may also be added.

[0077] The components included in the cosmetic composition may, in addition to the Trolox-peptide conjugate and carrier component of the present invention as active ingredients, further include, but are not limited to, components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0078] In another aspect, the present invention provides a method for preventing or treating hyperhidrosis, comprising the step of administering a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 to an individual (subject).

[0079] In another aspect, the present invention provides a pentapeptide comprising the amino acid sequence of Sequence ID No. 1 for use in the prevention or treatment of hyperhidrosis.

[0080] In another aspect, the present invention provides a method for suppressing body odor, which includes the step of administering a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 to an individual (subject).

[0081] In another aspect, the present invention provides a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 for use in suppressing body odor.

[0082] The term "subject" may include humans. Furthermore, the term "subject" may refer to an individual in need thereof, and such individual in need may include not only individuals diagnosed with hyperhidrosis or body odor, or individuals exhibiting symptoms related to hyperhidrosis or body odor, but also individuals who wish to receive the administration to prevent the onset of the aforementioned disease or symptoms or to improve their health.

[0083] The term "administration" means providing the specified substance to the patient by any appropriate method, and the administration route of the pentapeptide of the present invention may be orally or parenterally via any common route, as long as it can reach the target tissue. The pentapeptide may also be administered by any device that can move the active substance to target cells.

[0084] As described above, the pentapeptide of the present invention inhibits the excessive release of neurotransmitters by inhibiting the formation of SNARE complexes in nerve cells, and is highly effective in suppressing sweat production by penetrating to nerve cells in the muscle layer even when applied to areas where excessive sweating occurs. Therefore, it can be used as a raw material for pharmaceutical or cosmetic compositions aimed at preventing, improving, alleviating, or treating hyperhidrosis or suppressing body odor.

[0085] Another aspect of the present invention is to provide a pain-relieving composition comprising a pentapeptide consisting of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0086] In this specification, the term "peptide" means a linear or cyclic molecule formed by the bonding of amino acid residues to each other via peptide bonds. The preparation of such peptides may be achieved by conventional biological or chemical synthesis methods known in the art, such as solid-phase synthesis techniques.

[0087] In this specification, the term "pentapeptide" means a linear molecule consisting of five amino acid residues, and the pentapeptide of the present invention specifically means a linear peptide molecule composed of the amino acid sequence (KFLIK) of SEQ ID NO: 1.

[0088] The aforementioned "peptides" and "pentapeptides" may be amino acid variants or fragments having different sequences due to deletions, insertions, substitutions, or combinations thereof of amino acid residues, to the extent that they do not affect the function. Amino acid exchanges that do not alter the overall activity of the peptides are known in the art. In some cases, they may be modified by phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc. Accordingly, the present invention includes peptides and variants thereof, or active fragments thereof, having an amino acid sequence substantially identical to a pentapeptide composed of the amino acid sequence of KFLIK. The substantially identical proteins mean amino acid sequences having 75% or more, preferably 80% or more, for example, 85% or more, 90% or more, 95% or more, 98% or more, or 99% or more sequence homology with the amino acid sequence of KFLIK, but are not limited thereto; any protein having 75% or more amino acid sequence homology and the same activity is included in the scope of the present invention. Furthermore, the peptides of the present invention may additionally include targeted sequences, tags, labeled residues, or amino acid sequences manufactured for specific purposes to increase half-life or peptide stability.

[0089] Furthermore, protecting groups may be attached to the N-terminus or C-terminus of the peptide of the present invention to obtain better chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., broad biological activity spectrum), and reduced antigenicity. For example, the protecting group may be an acetyl group, a fluorenyl methoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, or polyethylene glycol (PEG), but may include any component that can modify the peptide, particularly enhance its stability. The term "stability" is used to include not only in vivo stability that protects the peptide of the present invention from attack by endogenous protein-cutting enzymes, but also storage stability (e.g., room temperature storage stability).

[0090] The pentapeptide composed of the amino acid sequence of KFLIK in this invention suppresses the expression of infectious cytokines in muscle cells and thereby suppresses inflammatory responses, making it possible to prevent or treat pain caused by micro-tears and the like.

[0091] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in the present invention can prevent or treat pain by suppressing the release of substance P, a pain transmission factor secreted from synaptic neurons, thereby preventing the transmission of pain.

[0092] Furthermore, the pentapeptide composed of the amino acid sequence of KFLIK in this invention penetrates to the muscle layer of skin tissue via transdermal absorption, so pain can be prevented, alleviated, or treated simply by applying it to the affected area.

[0093] In this invention, "pain" means an unpleasant sensation resulting from stimulation of specific nerves having pain receptors, and includes all symptoms in which such sensation is alleviated, prevented, or treated by the pentapeptide of the present invention. The pain according to the present invention may be acute or chronic pain.

[0094] In the present invention, the pain may be one or more selected from the group consisting of inflammatory pain, neuropathic pain, analgesic pain, and psychogenic pain, for example, cancer pain and postoperative pain in which analgesic pain, inflammatory pain, etc. act in combination; trigeminal neuralgia pain, idiopathic pain, and diabetic neuropathic pain which fall under neuropathic pain; and one or more selected from the group consisting of migraine and menstrual pain which fall under visceral pain.

[0095] Furthermore, compositions containing the pentapeptide of the present invention can be applied to pain occurring in the skin or musculoskeletal system, for example, diabetic neuropathy, diabetic peripheral neuropathy, pain due to viral infection, nociceptive pain, facial neuralgia, postherpetic neuralgia, lower back pain, disc pain, radiculopathy, neuropathy, hyperalgesia, and central nervous system sensitization. Sensitization-related pain, allodynia, cancer pain, myofascial pain syndrome, carpal tunnel syndrome, cubital tunnel syndrome (ulnar nerve disease), De Quervain's syndrome, rotator cuff syndrome, osteoarthritis, tendinitis, tenosynovitis, lateral humeral osteitis, medial humeral osteitis, tendon injury, bursitis, ligament inflammation, tennis elbow, golfer's elbow, knee pain, patellar tendinitis, meniscal cartilage injury, finger pain, wrist tendinitis, wrist sprain, ankle sprain, sprain, fibromyalgia, plantar fasciitis, inflammatory arthritis, infectious arthritis, rheumatoid arthritis, degenerative joint pain, frozen shoulder, gout, postpartum joint pain, osteomyelitis, ankylosing spondylitis Pain associated with one or more of the following conditions: spondylitis, degenerative vertebral disorders, lumbar spinal stenosis, cruralgia, sprains, contusions, joint pain, and sciatica.

[0096] Specifically, the pentapeptide according to the present invention can reach the muscle layer even when simply applied to the skin, and is absorbed through the skin, exhibiting excellent analgesic effects. Therefore, it can be particularly effective for pain in the skin or tissues located close to the skin, especially pain around areas with thin subcutaneous fat. For example, it can effectively prevent or treat sprains, bruises, joint pain, tendinitis, ligament inflammation, frozen shoulder, rotator cuff syndrome, and pain associated with tennis elbow, golfer's elbow, and De Quervain's syndrome in areas such as the knees, elbows, fingers, shoulders, ankles, and feet.

[0097] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of pain, comprising a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0098] In the present invention, the term "prevention" means all actions that suppress or delay pain by administering a composition containing the pentapeptide.

[0099] In the present invention, the term "treatment" means all actions in which pain is "relieved," "improved," or "modified" by the administration of the composition containing the pentapeptide.

[0100] The pharmaceutical compositions for the prevention or treatment of pain according to the present invention can be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, topical preparations, suppositories, and sterile injection solutions, respectively, by conventional methods, and may contain appropriate carriers, excipients, or diluents commonly used in the manufacture of pharmaceutical compositions for dosage formation.

[0101] The pharmaceutically acceptable carriers may include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, and others commonly used in formulation.

[0102] The pharmaceutical composition may further contain, in addition to the above-mentioned components, lubricants, humectants, sweeteners, flavoring agents, emulsifiers, suspension agents, preservatives, and the like.

[0103] The pharmaceutical composition may be administered orally or parenterally (e.g., intramuscularly, intravenously, intraperitoneally, subcutaneously, intradermally, or topically) by the method of intent, and the dosage will vary depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the time, but may be appropriately selected by those skilled in the art.

[0104] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective dose may be determined by factors including the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the elimination ratio, the duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other pain relievers, and may be administered simultaneously, separately, or sequentially with conventional therapeutic agents, and may be administered alone or in multiple doses. It is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, taking all of the above factors into consideration, and this can be easily determined by those skilled in the art.

[0105] The effective amount of the pharmaceutical composition may vary depending on the patient's age, sex, condition, weight, absorption rate of the active ingredient into the body, inactivation rate, excretion rate, type of disease, and concomitant drugs. It may also be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc. For example, the pharmaceutical composition may be administered at a dose of approximately 0.0001 μg to 500 mg, preferably 0.01 μg to 100 mg, per kg of the patient's body weight per day.

[0106] Another aspect of the present invention is the provision of a quasi-drug composition for the prevention or improvement of pain, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0107] The specific details of the peptide, pentapeptide, and pain are as described above.

[0108] In this invention, the term "quasi-drug" refers to articles used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, which have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Law, quasi-drugs are articles excluding those used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans / animals, and products that have a mild or no direct effect on the human body.

[0109] The quasi-drug composition of the present invention can be manufactured in one or more forms selected from the group consisting of body cleansers, foams, soaps, masks, ointments, creams, lotions, essences, and sprays, but is not limited thereto. Furthermore, the composition may be manufactured in the form of bands, sanitary napkins, etc., but is not limited thereto.

[0110] When the pentapeptide of the present invention is used as an additive for quasi-drugs, the composition can be added as is or used together with other quasi-drugs or quasi-drug components, and can be used appropriately by conventional methods.

[0111] In another aspect, the present invention provides a pain-relieving cosmetic composition comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0112] The specific details of the peptide, pentapeptide, and pain are as described above.

[0113] The cosmetic composition may be manufactured in any dosage form commonly produced in the art, such as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleanser, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited to these.

[0114] The aforementioned cosmetic composition may be manufactured in a variety of forms, such as softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, powder, hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, gel, or other solutions, sol-gels, emulsions, oils, waxes, and aerosols, but is not limited to these forms.

[0115] The cosmetic composition of the present invention may contain other additives such as excipients and carriers in addition to the pentapeptide composed of the amino acid sequence of KFLIK, and it is possible to apply and incorporate ordinary ingredients commonly used in general skin cosmetics in the required amounts.

[0116] If the dosage form of the cosmetic composition is a paste, cream, or gel, animal oils, vegetable oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as the carrier component.

[0117] When the dosage form of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier component, and in particular when it is a spray, propellants such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether may be additionally included, but are not limited to these.

[0118] If the dosage form of the cosmetic composition is a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as the carrier component. For example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan may be used.

[0119] If the dosage form of the cosmetic composition is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol, a suspension agent such as ethoxylated isostearyl alcohol, polyoxyethyl sorbitol ester, or polyoxyethylene sorbitan ester, or microcrystalline cellulose, aluminum methhydroxyl, bentonite, agar, or tragacanth.

[0120] If the dosage form of the cosmetic composition is a surfactant-containing cleanser, the carrier component may be an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinate monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0121] If the dosage form of the cosmetic composition is a hair shampoo, the pentapeptide conjugate of the present invention may be mixed with base components for forming the shampoo, such as a thickener, surfactant, viscosity modifier, humectant, pH adjuster, preservative, and essential oil. CDE may be used as the thickener, LES, an anionic surfactant, and cocobetaine, an amphoteric surfactant, may be used as the surfactant, polyquartz may be used as the viscosity modifier, glycerin may be used as the humectant, and citric acid and sodium hydroxide may be used as the pH adjuster. Grapefruit extract may be used as the preservative, and other essential oils such as cedarwood, peppermint, and rosemary, as well as silk amino acids, pentaol, or vitamin E may be added.

[0122] The components included in the cosmetic composition may, in addition to the pentapeptide and carrier component of the present invention as active ingredients, further include, but are not limited to, components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0123] In another aspect, the present invention provides a method for preventing or treating pain, comprising the step of administering a pentapeptide composed of the amino acid sequence of SEQ ID NO: 1 to an individual (subject).

[0124] In another aspect, the present invention provides a pentapeptide comprising the amino acid sequence of SEQ ID NO: 1 for use in the prevention or treatment of pain.

[0125] In another aspect, the present invention provides a quasi-drug composition for the prevention or improvement of pain, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0126] In another aspect, the present invention provides a pain-relieving cosmetic composition containing a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

[0127] The term "subject" may include humans. Furthermore, the term "subject" may refer to an individual in need of the pentapeptide of this application, and such individual in need of administration may include not only individuals diagnosed with pain or individuals experiencing pain, but also individuals who wish to receive the administration to prevent the onset of the disease or symptoms or to improve their health.

[0128] The term "administration" means providing a given substance to a patient by any appropriate method, and the administration route of the composition of the present invention may be orally or parenterally via any common route, as long as it can reach the target tissue. The composition may also be administered by any device that can move the active substance to target cells.

[0129] As described above, the pentapeptide of the present invention suppresses pain caused by inflammatory responses by inhibiting the expression of infectious cytokines in muscle cells, prevents pain transmission by inhibiting the release of substance P from nerve endings, and has excellent pain-suppressing effects even when applied to the skin, penetrating to nerve cells in the muscle layer. Therefore, it can be usefully used as a raw material for pharmaceuticals, quasi-drugs, or cosmetic compositions aimed at alleviating, improving, preventing, or treating pain.

[0130] The present invention will be described in detail below with reference to examples and experimental examples.

[0131] However, the following examples and experimental examples are for illustrative purposes only, and the content of the present invention is not limited to the following examples and experimental examples.

[0132] [Example 1] Synthesis and property verification of novel pentapeptides 1-1. Synthesis of pentapeptides A novel peptide sequence "KFLIK," consisting of the amino acid sequence described in Sequence ID No. 1, was prepared using a known method. The molecular weight of the peptide containing the amino acid sequence of Sequence ID No. 1 was measured using a molecular weight analyzer, and it was confirmed to be 647.4 Da.

[0133] 1-2. Evaluation of high-temperature stability during long-term storage The peptide of the present invention, composed of the amino acid sequence of Sequence ID No. 1, was dissolved in sterile distilled water at a concentration of 1000 ppm and stored at 45°C for 7, 14, 28, 60, and 75 days, after which HPLC analysis was performed.

[0134] As a result, as shown in Figure 1a, we were able to confirm that the peptide of the present invention maintains stability for 75 days, the maximum observation period, under conditions of 45°C.

[0135] 1-3. Evaluation of high-temperature stability The peptide of the present invention was dissolved in sterile distilled water at a concentration of 1000 ppm, heated at 121°C for 15 minutes and then 30 minutes, and then analyzed by HPLC.

[0136] As a result, as shown in Figure 1b, we were able to confirm that the peptide of the present invention maintained its stability for 30 minutes, the maximum heating time at 121°C.

[0137] [Experimental Example 1] Confirmation of the effect of inhibiting SNARE complex formation in nerve cells. 1-1. Confirmation of the effect of inhibiting SNARE complex formation To confirm the effect of the pentapeptide of the present invention on inhibiting neurotransmitter release in nerve cells, we investigated whether SNARE complex formation could be suppressed using a cell line derived from neuroblastoma cells (SH-SY5Y).

[0138] Specifically, 3 × 10⁶ SH-SY5Y cells 5Cells were inoculated into 6-well plates at a cell / well density and cultured for 24 hours in a 37°C, 5% CO2 incubator, after which the medium was replaced with serum-free medium. Subsequently, 6-well plates were treated with 10 μM, 50 μM, and 100 μM pentapeptides, and a positive control group was treated with 0.2 μM botulinum toxin type C light chain (BoNT / CLC) and cultured. After 24 hours, cells were collected, lysates were obtained, and Western blotting was performed against syntaxin 1a antibody (synapticsystems, Germany).

[0139] As a result, as shown in [Figure 2a], treatment with the pentapeptide of the present invention suppressed SNARE complex formation in a manner similar to that of the positive control group treated with botulinum toxin.

[0140] Through this process, it can be seen that the pentapeptide of the present invention inhibits the release of neurotransmitters by suppressing the formation of SNARE complexes and preventing synaptic vesicles from fusing with the presynaptic membrane.

[0141] 1-2. Confirmation of the effect of syntaxin 1α protein degradation To confirm the effect of the pentapeptide of the present invention on inhibiting neurotransmitter release in nerve cells, we investigated whether the pentapeptide of the present invention has the ability to degrade the syntaxin 1a protein that constitutes t-SNARE.

[0142] Specifically, the experimental group was treated with 1 μg of recombinant syntaxin 1a protein (Novus Biologicals, USA) and the pentapeptide of the present invention at concentrations of 20 μM, 100 μM, and 200 μM in reaction buffer (50 mM HEPES, 40 mM 2-ME, 20 μM ZnCl2, pH 7.4), while the negative control group (Control) was treated with recombinant syntaxin 1a protein only. After reacting at 37°C for 4 hours, Western blotting was performed using the syntaxin 1a antibody (synapticsystems, Germany).

[0143] As a result, as shown in [Figure 2b], it was confirmed that treatment with the pentapeptide of the present invention resulted in a concentration-dependent decrease in the recombinant syntaxin 1a protein band.

[0144] Through this process, it can be seen that the pentapeptide of the present invention has the effect of inhibiting neurotransmitter release by degrading the syntaxin 1a protein and preventing the formation of the SNARE complex.

[0145] [Experimental Example 2] Confirmation of the effect of inhibiting neurotransmitter release in nerve cells To confirm the effect of the pentapeptide of the present invention on inhibiting neurotransmitter release in nerve cells, the amount of acetylcholine, a neurotransmitter, secreted was measured using a cell line derived from neuroblastoma cells (SH-SY5Y).

[0146] Specifically, SH-SY5Y cells were injected into dishes and cultured for 24 hours in a 37°C, 5% CO2 incubator. The culture medium was then replaced with serum-free medium and cultured for another 48 hours. Subsequently, dishes were treated with 1 μM, 10 μM, and 50 μM pentapeptides, and a positive control group was cultured in dishes treated with 50 nM tetanus. After a certain period, nicotine (NIC) and potassium chloride (KCl) were added to induce acetylcholine release. The amount of acetylcholine secreted in the culture medium was then measured using an acetylcholine analysis kit.

[0147] As a result, as shown in [Figure 3], the release of acetylcholine increased in the groups treated only with nicotine (NIC) and potassium chloride (KCl) compared to the control group that received no treatment, but the release of acetylcholine was significantly suppressed in all groups treated with the pentapeptide of the present invention.

[0148] [Experimental Example 3] Confirmation of the effect of pentapeptide on suppressing hyperhidrosis through human efficacy evaluation. To confirm whether the pentapeptide according to the present invention actually has the effect of suppressing sweat secretion in the palms of the hands, soles of the feet, and armpits, we used potato starch and iodine to capture images of the amount of sweat secreted, and then performed intensity analysis using ImageJ software.

[0149] Specifically, the efficacy of the present invention on the degree of sweat secretion was evaluated in 10 subjects with localized hyperhidrosis (palms, soles of feet, armpits). A solution containing 2000 ppm of the present invention was applied to one side of the subject's palm, sole, or armpit using a mesolola. The opposite side of the palm, sole, or armpit was designated as a control group, and in the control group, PBS was applied with a mesolola instead of the pentapeptide solution. After sufficient absorption and natural drying, an iodine solution was applied to the same area using a sterile gauze pad and allowed to air dry. When the iodine solution was completely dry, corn starch was evenly applied to the iodine-coated area with cotton to form a starch layer. When sweat was secreted, the iodine and starch dissolved, causing a celadon color change, so the color change over time was observed on the subjects' palms, soles, or armpits. After image acquisition, intensity analysis was performed using ImageJ software.

[0150] As a result, as shown in [Figure 4], it was confirmed that the amount of sweat secreted from the hands of subjects who applied the lotion containing the pentapeptide of the present invention decreased by approximately 74%. Furthermore, as shown in [Figure 5], it was confirmed that the amount of sweat secreted from the soles of the feet of subjects who applied the lotion containing the pentapeptide of the present invention decreased by approximately 58%, and as shown in [Figure 6], it was confirmed that the amount of sweat secreted from the armpits of subjects who applied the lotion containing the pentapeptide of the present invention decreased by approximately 88%.

[0151] Through this, it can be seen that the pentapeptide of the present invention can provide therapeutic effects in patients with hyperhidrosis by reducing sweating and preventing excessive sweat production simply by applying it to the skin, thereby providing an effect that fundamentally blocks the generation of body odor.

[0152] [Experimental Example 4] Confirmation of intracellular or skin tissue penetration patterns 4-1. Preparation of Rhodamine-Pentapeptide Conjugate To confirm the intracellular penetration or tissue penetration pattern of the peptide of the present invention, a rhodamine-pentapeptide conjugate with a fluorescent substance was prepared. First, a 10 mg / ml pentapeptide solution was prepared using 100 mM sodium bicarbonate (pH 9.0), and a 1 mg / ml NHS-rhodamine (Thermo Scientific, 46406) solution was prepared using dimethylformamide. The prepared solutions were mixed so that the pentapeptide:NHS-rhodamine ratio was 1:10 (molar ratio). After shielding from light, the mixture was reacted at room temperature for 1 hour with inverting, and after dialyzing of the reaction product, the conjugation status was confirmed by LC / MS.

[0153] 4-2. Confirmation of intracellular osmosis To confirm whether the pentapeptide of the present invention penetrates into nerve cells, 3 × 10⁶ SH-SY5Y cells were used. 5 Cells were inoculated into 6-well plates at a cell / well density. After incubation for 24 hours in a 37°C, 5% CO2 incubator, the medium was replaced with serum-free medium. After treating the cells with rhodamine-peptide conjugates at different concentrations for 4 hours, the cells were fixed by treatment with 4% paraformaldehyde, and the nuclei were stained using a DAPI staining kit (Invitrogen, USA). Intracellular peptide penetration was observed via fluorescence microscopy. Blue indicates the nucleus of cells stained with DAPI, and red indicates the "rhodamine-pentapeptide" conjugate.

[0154] As a result, as shown in [Figure 7], the coexistence of blue and red colors confirmed that the pentapeptide of the present invention had penetrated into nerve cells.

[0155] 4-3. Confirmation of tissue penetration Seven-week-old SD rats were dehaired from the dorsal region, rhodamine-pentapeptide was applied, and the rats were sacrificed for analysis one hour later. Skin tissue from the application site was collected and fixed with formalin for one day. Paraffin blocks were prepared from the fixed tissue and sectioned, and then immunohistochemical staining was performed using TrkB Ab (Cell signaling, USA), a neuronal cell marker. Subsequently, nuclear staining was performed using a DAPI staining kit (Invitrogen, USA). The peptide penetration patterns within the tissue were observed via fluorescence microscopy. Blue indicates cell nuclei stained with DAPI, red indicates rhodamine-pentapeptide conjugates, and green indicates TrkB (nerve fiber marker).

[0156] As a result, as shown in [Figure 8], we confirmed that the pentapeptide of the present invention penetrated to the muscle layer of the skin tissue and that it co-localized with nerve cell markers.

[0157] Through this, the pentapeptide of the present invention can penetrate into nerve cells, penetrate to the muscle layer of skin tissue via transdermal absorption, and coexist with nerve cell markers in the muscle layer. Therefore, it can be shown that it is involved in the formation of the SNARE complex and can exert an effect of suppressing sweat secretion caused by excessive release of neurotransmitters, as well as an effect of blocking the generation of body odor caused by sweat secretion from the source.

[0158] Through this, the pentapeptide of the present invention can penetrate into nerve cells and penetrate to the muscle layer of skin tissue via transdermal absorption. Since it coexists with nerve cell markers in the muscle layer, it can be shown to be involved in the suppression of infectious cytokine expression in muscle cells and the release of pain transmission factors at nerve endings, thereby exhibiting an effect of suppressing the occurrence and transmission of pain.

[0159] [Experimental Example 5] Confirmation of the body odor-suppressing effect of pentapeptides using body odor-inducing bacterial strains. The antibacterial effect of the pentapeptide of the present invention was confirmed using Corynebacterium, a representative odor-causing bacterium known to break down substances in sweat and induce unpleasant odors.

[0160] Specifically, Corynebacterium xerosis (C. xerosis, KCTC9105), a strain that induces body odor, was purchased from the Bioresource Center of the Korea Institute of Biotechnology and Biotechnology and used. The strain was inoculated into Brain Heart Infusion (Difco, USA) medium and cultured in a 37°C incubator for 2 days. The same medium was added to the cultured strain, and 10 6 A dilution prepared to a concentration of CFU / ml was used as the test bacterial suspension. 100 μL of the test bacterial suspension was separated into 96-well plates, and then treated with pentapeptides at different concentrations. For the positive control group, 1% triclosan was added instead of pentapeptide. After 48 hours of incubation, absorbance was measured at 600 nm using a spectrophotometer to confirm the effect of inhibiting the growth of odor-causing bacteria.

[0161] As a result, as shown in [Figure 9], it was confirmed that treatment with the pentapeptide of the present invention clearly suppressed the growth of body odor-inducing bacterial strains. In particular, the groups treated with 1 mg and 2 mg of pentapeptide showed excellent antibacterial effects at a level comparable to the positive control group.

[0162] Through this, it can be seen that the pentapeptide of the present invention can effectively suppress the characteristic body odor that occurs when sweating by inhibiting the growth of body odor-inducing bacterial strains.

[0163] [Experimental Example 6] Confirmation of the effect of pentapeptide treatment on suppressing infectious cytokine release in muscle cells. 6-1. Confirmation of the expression levels of infectious cytokine genes To confirm the pain-improving effect of the pentapeptide of the present invention, the expression of inflammation-related cytokines was examined using mouse myoblasts. Infectious cytokines are involved in pain in neurological disorders, and when infectious cytokines such as TNF-α are expressed after nerve injury, an inflammatory response is triggered, causing pain.

[0164] Specifically, 2 × 10⁶ mouse myoblast cells, C2C12 cells 5 Cells were inoculated into 6-well plates at a density of cells / well and cultured in DMEM (10% PBS) medium for 3 days. When the cells had grown to about 70-80% of the plate's capacity, they were switched to differentiation medium (DMEM, 5% horse serum) and cultured for 3 days. Subsequently, they were switched to differentiation medium (DMEM, 2% horse serum) and cultured for 4 days to induce differentiation from myoblasts to myotubes. Then, they were cultured in serum-free medium for 2 hours and pretreated with pentapeptides at concentrations of 1 μM, 10 μM, and 100 μM for 1 hour. After that, they were treated with 20 nM TNF-α and cultured for 24 hours to induce an inflammatory response.

[0165] Subsequently, cells were collected and RNA was separated, and cDNA was synthesized using a cDNA synthesis kit (Intron, Korea). Polymerase chain reaction was performed using the synthesized cDNA, PCR premix (Intron, Korea), and IL-1α, TNF-α, IL-1β, and COX-2 primers. The amplified PCR products were subjected to electrophoresis on an agarose gel, and the degree of mRNA expression of the aforementioned genes was compared by confirming the DNA bands. The primer sequences used are shown in [Table 1].

[0166] [Table 1]

[0167] As a result, as shown in [Figure 10a], when inflammation was induced in myotubes by treating with TNF-α, the mRNA expression levels of IL-1α, IL-1β, COX-2, and TNF-α, i.e., infectious cytokines, were significantly increased compared to the untreated group (control). However, in the 1 μM and 10 μM pentapeptide treatment groups of the present invention, the increased expression of infectious cytokines was reduced.

[0168] 6-2. Confirmation of the expression level of infectious cytokine proteins On the other hand, to confirm the expression levels of inflammatory cytokine proteins, some of the cells recovered in 2-1 were subjected to (WB) cell lysate separation, and the proteins were quantified and mixed with sample buffer to prepare the samples. These samples were then spread on SDS-PAGE and nitrocellulose membranes and blocked with 5% scheme milk. COX-2 antibody (Cell Signaling Technology, USA), IL-1β antibody (Cell Signaling Technology, USA), and TNF-α antibody (Cell Signaling Technology, USA) were primarily bound, followed by secondary antibody binding. After processing with ECL solution, the expression levels of the target proteins were observed.

[0169] As a result, as shown in [Figure 10b], when inflammation was induced in myotubes by treating with TNF-α, the levels of protein expression of COX-2, IL-1β, and TNF-α, i.e., the amount of infectious cytokine proteins, were significantly increased compared to the untreated group (control). However, in the pentapeptide-treated group of the present invention, the increased expression of infectious cytokines was found to decrease in a concentration-dependent manner.

[0170] Through this process, the pentapeptide of the present invention exhibits the effect of reducing the expression of IL-1α, IL-1β, COX-2, and TNF-α in myotubes, i.e., the expression of infectious cytokines, at both the mRNA and protein levels. Therefore, it can be usefully used as a substance for preventing or treating pain.

[0171] [Experimental Example 7] Confirmation of the inhibitory effect on neurotransmitter release in nerve cells To confirm the pain-relieving effect of the pentapeptide of the present invention, the degree of release of substance P at synapses was examined using mouse nerve cells. Since substance P transmits pain from the periphery to the central nervous system, suppressing the release of substance P may result in pain relief.

[0172] Specifically, the dorsal root ganglion (DRG) was isolated from the spinal cord of 6-week-old rats, washed, and then the nerve cells were isolated. The isolated nerve cells were divided into 24-well plates and inoculated, then cultured for 1 day. After that, a similar mitotic inhibitor was mixed into the culture medium, and only pure nerve cells were cultured for 6 days. Then, the medium was changed to B-27 medium (serum-free, 1% penicillin) to add nutrients and cultured for 24 hours, followed by pretreatment with 1 μM and 10 μM pentapeptides for 1 hour. Subsequently, 100 μM capsaicin was treated for 30 minutes to induce the release of substance P, and the amount of substance P released was analyzed by ELISA.

[0173] As a result, as shown in [Figure 11], treatment with capsaicin increased the release of substance P compared to the untreated control group, but in the case of the pentapeptide-treated group of the present invention, it was confirmed that the increased release of substance P was significantly reduced.

[0174] Through this, it can be seen that the pentapeptide of the present invention can be usefully used as a pain preventive or therapeutic substance by suppressing the release of substance P in nerve cells and preventing the transmission of pain signals.

[0175] [Experimental Example 8] Confirmation of the pain-relieving effect of pentapeptides through human efficacy evaluation. To confirm whether the pentapeptide according to the present invention actually exhibits a pain-suppressing effect, delayed-onset muscle pain was induced in 20 subjects, and then a lotion containing the pentapeptide according to the present invention was applied to the painful area to evaluate the degree of pain reduction in the human body.

[0176] Specifically, to induce delayed-onset muscle soreness (DMS), subjects were instructed to stand with their feet shoulder-width apart and their backs straight. They then performed a series of 2-minute intervals of lifting and lowering their heels, followed by 3-minute rests. This exercise was repeated for a total of 3 sets. After 24 hours, a solution containing 2000 ppm of the pentapeptide of the present invention was applied to the painful area. Subsequently, at 24, 48, and 72 hours, subjects self-measured their pain levels using a Numeric Rating Scale (NRS) while walking (0 representing no pain, 10 representing the worst pain imaginable). The mean PID scores were calculated using the following formula, and after the 72-hour trial, the degree of pain reduction was confirmed through a subject questionnaire.

[0177] [Mathematical formula] Pain Intensity Difference (PID) = Pain intensity after application of the pentapeptide of the present invention - Pain intensity before application of the pentapeptide of the present invention

[0178] As a result, as shown in [Figure 12a] and [Figure 12b], the degree of pain induced 24 hours after exercise was significantly improved 72 hours after exercise (48 hours after lotion application) in the experimental group treated with the pentapeptide of the present invention.

[0179] Through this, it can be seen that the pentapeptide of the present invention can provide a therapeutic effect in reducing pain even when simply applied to the skin of a patient experiencing muscle pain.

Claims

1. A pain-relieving composition comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient, wherein the pain is one or more selected from the group consisting of inflammatory pain, postoperative pain, cancer pain, trigeminal neuralgia pain, idiopathic pain, diabetic neuropathic pain, myalgia, and menstrual pain.

2. The composition according to claim 1, wherein the pentapeptide i) suppresses the expression of one or more infectious cytokines selected from the group consisting of IL-1α, IL-1β, COX-2, and TNF-α, and ii) suppresses the secretion of substance P (substance P), which is a pain transmission factor.

3. The composition according to claim 1, wherein the pain is pain associated with any one selected from the group consisting of sprains, contusions, joint pain, tendinitis, ligament inflammation, frozen shoulder, rotator cuff syndrome, tennis elbow, golfer's elbow, and De Quervain's syndrome of the knee, elbow, fingers, shoulder, ankle, or foot.

4. A pharmaceutical composition for the prevention or treatment of pain, comprising a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient, wherein the pain is one or more selected from the group consisting of inflammatory pain, postoperative pain, cancer pain, trigeminal neuralgia pain, idiopathic pain, diabetic neuropathic pain, myalgia, and menstrual pain.

5. The pharmaceutical composition according to claim 4, wherein the pain is pain associated with any one selected from the group consisting of sprains, contusions, joint pain, tendinitis, ligament inflammation, frozen shoulder, rotator cuff syndrome, tennis elbow, golfer's elbow, and De Quervain's syndrome of the knee, elbow, fingers, shoulder, ankle, or foot.

6. A cosmetic composition for pain relief containing a pentapeptide composed of the amino acid sequence of Sequence ID No. 1 as an active ingredient.

7. The cosmetic composition for pain relief according to claim 6, wherein the cosmetic composition is in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, or spray.

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