KR20210136579A - Composition for Suppressing Pain Containing BigLEN - Google Patents

Abstract

The present invention relates to a composition for inhibiting pain, and more particularly, to a pharmaceutical composition for inhibiting pain or preventing pain comprising a peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 as an active ingredient. According to the present invention, BigLEN exerts an inhibitory effect on formalin-induced pain models and CFA-induced inflammatory hyperalgesia, mechanical hyperalgesia, thermal hyperalgesia, and neuropathic pain, thereby being able to be used for pain suppression and pain prevention.

Description

Composition for suppressing pain containing BigLEN {Composition for Suppressing Pain Containing BigLEN}

The present invention relates to a composition for inhibiting pain, and more particularly, to a pharmaceutical composition for inhibiting pain or preventing pain comprising the peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

Pain is a serious disease that reduces both the quality of life of an individual and social productivity. Pain is defined as an unpleasant sensory and emotional experience associated with actual or potential tissue damage. Although pain appears as a major symptom in various diseases, its perception is very subjective, so it is one of the most difficult pathologies to effectively diagnose and treat. Pain induces severe impairment of functional abilities and jeopardizes the patient's work, social and family life.

These pains vary widely, including inflammatory pain, neuropathic pain, osteoarthritis pain, postoperative pain, cancer pain, pain associated with metastatic cancer, pain induced by cancer chemotherapy agents, trigeminal neuralgia, burning pain, acute herpes and herpes. These include posterior neuralgia, laryngeal neuralgia, sympathetic dystrophy, fibromyalgia, gout pain, burn pain, phantom pain, complex regional pain syndrome, idiopathic pain syndrome, migraine, and chronic pain of unknown cause.

There are two main types of pain relievers currently in use. The first is a group consisting of nonsteroidal anti-inflammatory drugs (NSAIDs) and related COX-2 inhibitors, and the second is opioids such as morphine (Romsing J1 and Moiniche S., Acta Anaesthesiol Scand., 48(5):525). , 2004). These analgesics are effective in suppressing normal reactions. However, they have little effect on some types, such as neuropathic pain. However, some analgesic effect can be obtained by administering a high dose of opioids, but there is a possibility of serious side effects or poisoning due to an increase in dose. NSAIDs, which have a lower analgesic effect than opioids, require a higher dose to be effective, but also have the problem of causing gastrointestinal side effects unique to NSAIDs.

Accordingly, the present inventors made diligent efforts to develop a pain inhibitor that exhibits alleviating effects on both inflammatory hyperalgesia and neuropathic hyperalgesia. It was confirmed that it shows, and the present invention was completed.

It is an object of the present invention to provide a pharmaceutical composition for pain suppression or pain prevention.

Another object of the present invention is to provide a health functional food for pain suppression or pain prevention.

In order to achieve the above object, the present invention provides a peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, diastereomers, enantiomers, and amino acid substitutions selected from the group consisting of orthologs. Provided is a pharmaceutical composition for pain suppression or pain prevention comprising a peptide as an active ingredient:

<SEQ ID NO: 1>

LENPSPEAPARRRLLPP.

The present invention also provides a peptide selected from the group consisting of a peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, a diastereomer, an enantiomer, and an amino acid substitution ortholog thereof as an active ingredient. Provided is a functional health food for pain suppression or pain prevention comprising:

<SEQ ID NO: 1>

LENPSPEAPARRRLLPP.

According to the present invention, BigLEN exerts an inhibitory effect on formalin-induced pain models and CFA-induced inflammatory hyperalgesia, mechanical hyperalgesia, thermal hyperalgesia and neuropathic pain, and thus can be used for pain suppression and pain prevention.

1 shows the effect of BigLEN in a formalin-induced pain model. The test compound was administered intrathecally (a) and subcutaneously in the plantar (b) 1 hour before 20 μl of 5% formalin was injected into the sole of the foot, respectively. The positive control compound was intrathecally administered using gabapentin and pregabalin (1a). The duration of licking/biting of the injected paw during the early and late phases was recorded. The results are presented as the mean of 5 mice per group ± SEM. ***P<0.001 was significantly different from vehicle treated control.
Figure 2 shows the results of confirming the effect of BigLEN on inflammatory mechanical hyperalgesia in the CFA inflammatory pain model, and the foot lift threshold for mechanical stimulation was measured in mice. Base line (BL) measurements were obtained prior to intraplantar injection of CFA into mice and results are presented as the average of 5 mice per group. After CFA injection, BigLEN was administered intrathecally (a) subcutaneously on the sole of the foot (b), respectively. *P<0.05, **P<0.01, and ***P<0.001 were treated with excipients at the same time point. significantly different from the control group.
3 shows the effect of BigLEN on inflammatory thermal hyperalgesia in the CFA inflammatory pain model. Pain responses to thermal stimuli were measured in mice using a Hargreaves device. Base line (BL) measurements were obtained prior to intraplantar injection of CFA into the mice, and the results are presented as the average of 5 mice per group. After CFA injection, the results of administration of BigLEN intrathecally (a) subcutaneously on the soles of the feet (b) are shown. **P<0.01, and ***P<0.001 were significantly different from vehicle-treated controls at the same time point.
4 shows the effect of BigLEN on mechanical hyperalgesia in a neuropathic pain model. In a mouse model in which neuropathy was induced by surgical treatment, the threshold for lifting the foot to mechanical stimulation was measured in mice, and baseline (base line or BL) measurements were obtained before the surgical operation. After measurement on the 14th day of surgery, BigLEN was administered intrathecally (a) subcutaneously on the sole of the foot (b). **P<0.01, and ***P<0.001 were significantly different from vehicle-treated controls at the same time point.
5 shows the effect of BigLEN on thermal hyperalgesia in a neuropathic pain model, and the pain response to thermal stimulation was measured in mice using a Hargreaves device. Thresholds for thermal stimulation were measured in a mouse model inducing neuropathy by surgical treatment, and baseline (or BL) measurements were obtained before surgery. It was administered intrathecally (a) subcutaneously on the sole of the foot (b). **P<0.01, and ***P<0.001 were significantly different from vehicle-treated controls at the same time point.

In the present invention, as a result of trying to find a new pain-suppressing substance for which no pain-suppressing effect was known, it was confirmed that BigLEN derived from ProSAAS, a neuropeptide abundant in the brain, exhibited excellent pain-suppressing effect in a mouse pain model.

Accordingly, in one aspect, the present invention provides a peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, a diastereomer, an enantiomer, and an amino acid substitution peptide selected from the group consisting of orthologs. It relates to a pharmaceutical composition for pain suppression or pain prevention comprising as an active ingredient:

<SEQ ID NO: 1>

LENPSPEAPARRRLLPP

or Leu-Glu-Asn-Pro-Ser-Pro-Glu-Ala-Pro-Ala-Arg-Arg-Leu-Leu-Pro-Pro.

In the present invention, the amino acid constituting the amino acid sequence of SEQ ID NO: 1 may be characterized in that it is an L- amino acid.

In the present invention, the centromere may be characterized in that it has one or more amino acid substitutions selected from the group consisting of N3T, S5A, E7Q and A8V in the amino acid sequence of SEQ ID NO: 1.

In the present invention, BigLEN is a peptide represented by the amino acid sequence of Formula 1 or SEQ ID NO: 1.

<Formula 1>

L-leucyl-L-alpha-glutamyl-L-asparagyl-L-prolyl-L-seryl-L-prolyl-L-glutaminyl-L-alanyl-L-prolyl-L-alanyl-L-arginyl-L-arginyl- L-leucyl-L-leucyl-L-prolyl-L-proline

In the present invention, the amino acid sequence of SEQ ID NO: 1 is that of a mouse-derived BigLEN, and in the case of a human, it has the BigLEN of the following sequence as a centromere.

LETPAPQVPARRLLPP-human (SEQ ID NO: 2)

(Leu-Glu-Thr-Pro-Ala-Pro-Gln-Val -Pro-Ala-Arg-Arg-Leu-Leu-Pro-Pro)

BigLEN used in the present invention includes centromeres other than the peptide having the amino acid sequence of SEQ ID NO: 1.

In the present invention, "ortholog" refers to a gene (or protein) having the same function that exists between different species. These are genes that evolved from a common ancestral gene during speciation (the process of creating a species) and generally retain the same function during the evolutionary process.

In the present invention, the BigLEN is L-asparagine 3, L-serine 5, 7-glutamic acid, and L-alanine 8 from the amino acid sequence (SEQ ID NO: 1), respectively, L-threonine, L-alanine, L It can be replaced with a BigLEN centromere having an amino acid sequence substituted with -glutamine and L-valine.

In addition, the BigLEN of the present invention can be formed with an acceptable salt in preparation with both organic and inorganic acids or bases. For example, acid addition salts of base compounds are prepared by dissolving the organic base in either an aqueous solution or an aqueous alcoholic solution, or other suitable solvent comprising a suitable acid, and evaporating the solution to isolate the salt. Examples of preparations acceptable salts are chloride, bromide, sulfate, hydrochloride, hydrobromide, hydrosulfate and the like, as well as salts of sodium, potassium and magnesium and the like.

The pain targeted by the present invention includes inflammatory pain, neuropathic pain, osteoarthritis pain, postoperative pain, cancer pain, pain associated with metastatic cancer, pain caused by cancer treatment chemotherapeutic agents, trigeminal neuralgia, burning pain, acute herpes and Postherpetic neuralgia, laryngeal neuralgia, sympathetic dystrophy, fibromyalgia, gout pain, burn pain, phantom pain, complex regional pain syndrome, idiopathic pain syndrome, migraine, chronic pain of unknown cause, and the like. Neuropathic pain is caused by injury or infection of peripheral sensory nerves. Examples of specific causes of neuropathic pain include peripheral nerve trauma, herpes virus infection, diabetes, brachial plexus enucleation, neuroma, amputation and vasculitis, chronic alcoholism, human immunodeficiency virus infection, hypothyroidism, uremia, or vitamin deficiency. Including, but not limited to, nerve injuries caused by

When an effective amount of the compound of Formula 1 is used together with a carrier that is acceptable for administration to humans and mammals, the compound can be used as a pharmaceutically acceptable composition for the treatment of pain. It can be used in a method of alleviating a disorder resulting from pain in a mammal, including a human, by administering it to humans and mammals suffering from the pain listed above.

The compounds of the present invention may be prepared in various dosage forms and may be administered by oral routes or parenteral routes such as intravenous, intramuscular, subcutaneous, or intrathecal injection. For example, compositions thereof can be prepared in an inert, pharmaceutically acceptable carrier, either solid or liquid. That is, according to conventional methods, powders, powders, tablets, dispersible granules, capsules, suspensions, emulsions, syrups, etc. can be formulated and used in the form of oral dosage forms, external preparations, suppositories, and sterile injection solutions. In addition, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents or cataplasmas can be prepared and used as external compositions, but the present invention is not limited thereto. In addition, as the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used, but the present invention is not limited thereto.

The composition according to the present invention may further include a commonly used carrier, excipient, disintegrant, sweetener, lubricant, flavoring agent and diluent. The carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The disintegrant includes sodium starch glycolate, crospovidone, croscarmellose sodium, alginic acid, carboxymethyl cellulose calcium, carboxymethyl cellulose sodium, chitosan, guar gum, low-substituted hydroxypropyl cellulose, magnesium aluminum silicate, polacrylline potassium, etc.

In addition, the pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, Calcium hydrogen phosphate, lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, Aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol and the like may be used.

The amount of active compound in the dosage form of a unit dose may vary, or may be adjusted to about 0.001 mg to about 1 g per dose, based on an average human weight of 70 kg, preferably 0.01 to 100 mg, more preferably can be adjusted to 0.1 to 100 mg.

However, the dosage may vary depending on the requirements of humans and mammals, the severity of the disease to be treated and the compound used. It is within the skill of the artisan to determine the appropriate dosage for a particular situation. In addition to BigLEN, the composition of the present invention may contain at least one active ingredient that exhibits a similar pain-suppressing function.

In addition, the present invention provides a method for inhibiting pain comprising administering the composition for inhibiting pain to an individual. The subject is a vertebrate, preferably a mammal, more preferably an animal such as a rat, rabbit, guinea pig, hamster, dog, or cat, and most preferably a simian animal such as a chimpanzee or a gorilla. In addition, the administration method may be oral or parenteral administration, and when administered parenterally, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intrauterine dural injection, intracerebroventricular injection or chest It may be administered by intravenous injection. In addition, the composition for inhibiting pain may be used alone or in combination with methods using surgery, hormone therapy, drug therapy, and biological response modifiers to suppress pain.

The preferred dosage of the composition of the present invention varies depending on the condition and weight of the patient, the severity of the disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. Administration may be administered once a day, or may be administered in several divided doses.

In another aspect, the present invention provides an effective peptide selected from the group consisting of the peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, diastereomers, enantiomers and amino acid substitution orthologs thereof. It relates to a health functional food for pain suppression or pain prevention, including as an ingredient:

<SEQ ID NO: 1>

LENPSPEAPARRRLLPP.

In the present invention, the amino acid constituting the amino acid sequence of SEQ ID NO: 1 may be characterized in that it is an L- amino acid.

In the present invention, the centromere may be characterized in that it has one or more amino acid substitutions selected from the group consisting of N3T, S5A, E7Q and A8V in the amino acid sequence of SEQ ID NO: 1.

The BigLEN of the present invention may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of its use (for prevention or improvement). In general, in the production of food or beverage, the BigLEN of the present invention is added in an amount of 0.2 to 20% by weight, preferably 0.24 to 10% by weight, based on the raw material. However, in the case of long-term ingestion for health and hygiene or health control, the amount may be less than or equal to the above range.

The health food of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is preferably selected in the range of 0.01 to 0.04 parts by weight, preferably about 0.02 to 0.03 parts by weight, per 100 parts by weight of the health food of the present invention.

There is no particular limitation on the type of the food. Examples of foods to which BigLEN can be added include drinks, sausages, chocolates, candies, snacks, sweets, gums, dairy products including ice cream, beverages, alcoholic beverages, and vitamin complexes. include all

In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not very important, but is generally selected in the range of 0.01 - 0.1 parts by weight per 100 parts by weight of the health food of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1: Effect of BigLEN in the formalin-induced pain model

The effect of BigLEN was derived by conducting a pain test using the formalin-induced pain model, which is a kind of standardized pain model.

Male C57BL/6 mice (20 - 25 g) were tamed for at least 30 minutes prior to testing in an observation windshield chamber. A 5% formalin solution prepared in 20 μl isotonic saline was subcutaneously injected into the plantar surface of the left hind paw of the mouse, confirming that the mouse started the formalin-induced pain response, hind paw licking and biting. Immediately after formalin injection, licking and biting of the injected hind paws were recorded at 5-minute intervals for 60 minutes. Results were expressed as the mean of licking and biting times during the early phase of general pain response (0-10 minutes) and late phase of diseased pain response (10-45 minutes).

One hour before formalin injection, BigLEN (Tocris Bioscience, 125 μg/kg) was administered subcutaneously or intrathecally to the corresponding plantar plantar. The positive control compound was intrathecally administered using 10 mg/kg of gabapentin (gabapentin, Sigma-Aldrich) and 100 mg/kg of pregabalin (Sigma-Aldrich).

As a result, as shown in FIG. 1 , when BigLEN was administered intrathecally ( FIG. 1A ) and subcutaneously on the sole of the foot ( FIG. 1B ), it was confirmed that the licking/biting behavior of the late formalin reaction was inhibited.

In particular, there was no effect in the early stage and there was a statistically significant inhibitory effect in the later stage, suggesting that BigLEN is effective in alleviating disease-related pain, not pain as an essential sensation required in daily life.

Example 2: Effect of BigLEN on complete Freund's adjuvant (CFA)-induced inflammatory hyperalgesia

The effect of BigLEN was derived by conducting a pain test using the CFA-induced inflammatory pain model, which is a kind of standardized pain model.

Prior to intraplantar administration of 10 μl CFA (Thermofisher) to one hind paw of male C57BL/6 mice (20 - 25 g), the threshold of lifting to mechanical stimulation was measured, and baseline measurements were obtained. Footpad inflammation was induced by administration of 10 μl CFA, and thus BigLEN (1.25 μg/kg, 12.5 μg/kg and 125 μg/kg) was administered to male C57BL/6 mice 24 and 48 hours after the onset of apical hyperalgesia. administered. Using these mice, the effect of BigLEN on inflammatory hyperalgesia was measured.

First, the effect of BigLEN on mechanical hyperalgesia was evaluated using von Frey filaments (stoelting, Illinois, USA). Mice were placed in iron net floor cages to allow access to the filaments on the paws. Before starting the experiment, the mice were first acclimatized to the environment. The plantar surface of the mouse hindpaw was stimulated with Von Frey filaments while increasing the intensity of the force until a foot-lifting response, which is a pain response, was induced. Each Von Frey filament was applied to the paw for about 5 seconds. When a foot-lifting response occurred by a filament stimulation of a certain strength, the strength of the force of the Von Frey filament was lowered in the following order, and the foot stimulation was retested until the response did not occur. In the absence of a foot-lifting response, the strength of the Von Frey filament was increased and the foot stimulation was retested until a response occurred. The strength of the force required to trigger the foot-lift response is the nociceptive threshold for detecting mechanical pain. When this threshold was statistically significantly low, it was judged that mechanical hyperalgesia had occurred, and when this threshold was statistically significantly increased compared to the hyperalgesia threshold, it was judged that hyperalgesia was alleviated.

After administration of BigLEN (1.25μg/kg, 12.5μg/kg and 125μg/kg) subcutaneously or intrathecally, the mechanical hyperalgesia threshold as above was measured at 1 hour and 2 hours after administration. As a result, As shown in FIG. 2 , when BigLEN was administered subcutaneously to the sole of the foot ( FIG. 2B ) or intrathecally ( FIG. 2A ), it was confirmed that mechanical hyperalgesia was suppressed.

In addition to the effects of the previous mechanical hyperalgesia, the effects of BigLEN on thermal hyperalgesia were evaluated. Paw Withdrawal Latency (PWL) latency baseline, which is a pain response according to heat stimulation, was obtained for each mouse using the Hargreaves model, which is a heat stimulation pain induction model. In order to induce hyperalgesia by standard inflammation, CFA was injected as described above. 24 and 48 hours after CFA administration, BigLEN (1.25 μg/kg, 12.5 μg/kg, and 125 μg/kg) was administered subcutaneously or intrathecally on the plantar surface to reduce the above-mentioned delay time of lifting the foot at 2 hours and 2 hours. measured.

As a result, as shown in FIG. 3 , a statistically significant decrease in PWL caused by CFA-induced inflammation, that is, thermal hyperalgesia, was statistically significantly inhibited by BigLEN treatment.

Example 3: Effect of BigLEN on Neuropathic Pain

The standard neuropathic pain induction model was carried out on mice by adopting the method disclosed in the literature by Bennett et al . in 1988 (Bennett et al., Pain , 33: 87-107, 1988). In short, it is a model that induces hyperalgesia from 3 days after inducing neuropathy by damaging the sciatic nerve of the hind leg, and causing chronic pain after about 2 weeks. In this example, the effect of BigLEN was evaluated in the corresponding neuropathic pain model.

Male C57BL/6 (20-25 g) was used for all experiments, and prior to surgical intervention to induce neuropathy, mice were housed in a 12-hour day/night cycle environment with food and water ad libitum. Surgical treatment was performed on the mouse, and anesthesia was maintained using a _{2% isofluorane and 1.4 O 2} /NO _{2 mixture during the treatment.} The hairs on the thigh area of the hind legs were shaved and the surface was disinfected with iodine. Expose the sciatic nerve between the muscles to the outside by making an incision in the skin and fascia of the affected area. After minimizing drying in the air by providing isotonic saline, loosen the entire sciatic nerve 3 times with a silk thread. tied up The sciatic nerve was then placed in place and the fascia and skin were sutured. After this surgical procedure, mice showing no signs of infection at the wound site were used for evaluation, and the effect of BigLEN on hyperalgesia after 14 days of surgical treatment was evaluated. All mice used for evaluation during the experiment were in good health.

First, the effect of BigLEN on mechanical hyperalgesia was evaluated using von Frey filaments (stoelting, Illinois, USA). Mice were placed in iron net floor cages to allow access to the filaments on the paws. Before starting the experiment, the mice were first acclimatized to the environment. The plantar surface of the mouse hindpaw was stimulated with Von Frey filaments while increasing the intensity of the force until a foot-lifting response, which is a pain response, was induced. Each Von Frey filament was applied to the paw for about 5 seconds. When a foot-lifting response occurred by a filament stimulation of a certain strength, the strength of the force of the Von Frey filament was lowered in the following order, and the foot stimulation was retested until the response did not occur. In the absence of a foot-lifting response, the strength of the Von Frey filament was increased and the foot stimulation was retested until a response occurred. The strength of the force required to trigger the foot-lift response is the nociceptive threshold for detecting mechanical pain. When this threshold was statistically significantly low, it was judged that mechanical hyperalgesia had occurred, and when this threshold was statistically significantly increased compared to the hyperalgesia threshold, it was judged that hyperalgesia was alleviated. After administration of BigLEN (1.25 μg/kg, 12.5 μg/kg and 125 μg/kg) subcutaneously or intrathecally, the mechanical hyperalgesia threshold as above was measured at 1 hour and 2 hours after administration.

As a result, as shown in FIG. 4 , it was confirmed that mechanical hyperalgesia was suppressed by the administration of BigLEN (1.25 μg/kg, 12.5 μg/kg and 125 μg/kg).

In addition to the effects of the previous mechanical hyperalgesia, the effects of BigLEN on thermal hyperalgesia were evaluated. Paw Withdrawal Latency (PWL) latency baseline, which is a pain response according to heat stimulation, was obtained for each mouse using the Hargreaves model, which is a heat stimulation pain induction model. In order to induce hyperalgesia by standard inflammation, CFA was injected as described above. 24 hours and 48 hours after CFA administration, BigLEN 1.25 μg/kg, 12.5 μg/kg, and 125 μg/kg) were administered subcutaneously or intrathecally on the soles of the feet to measure the above-mentioned delay time of lifting the feet at the time and 2 hours. As a result, as shown in FIG. 5 , a statistically significant decrease in PWL caused by CFA-induced inflammation, that is, thermal hyperalgesia, was statistically significantly inhibited by BigLEN treatment.

statistics

In the present invention, the data of the formalin-induced pain model of Example 1 (FIG. 1) was tested for statistical significance using Student's T-test, and mechanical hyperalgesia and thermal hyperalgesia data of other pain models (FIG. 4~ 5) was tested for statistical significance by applying one-way ANOVA.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. will be. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

<110> Korea University Research & Business Foundation <120> Composition for Suppressing Pain Containing BigLEN <130> P19-B125 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> mouse BigLEN <400> 1 Leu Glu Asn Pro Ser Pro Glu Ala Pro Ala Arg Arg Leu Leu Pro Pro 1 5 10 15 <210> 2 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> human BigLEN <400> 2 Leu Glu Thr Pro Ala Pro Gln Val Pro Ala Arg Arg Leu Leu Pro Pro 1 5 10 15

Claims (8)

The peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, a diastereomer, an enantiomer, and a peptide selected from the group consisting of amino acid substitution orthologs as an active ingredient for pain suppression or a pharmaceutical composition for preventing pain:
<SEQ ID NO: 1>
LENPSPEAPARRRLLPP.
The composition according to claim 1, wherein the amino acids constituting the amino acid sequence of SEQ ID NO: 1 are L-amino acids.
The composition according to claim 1, wherein the centromere has one or more amino acid substitutions selected from the group consisting of N3T, S5A, E7Q and A8V in the amino acid sequence of SEQ ID NO: 1.
According to claim 1, wherein the pain is inflammatory pain, neuropathic pain, osteoarthritis pain, postoperative pain, cancer pain, pain associated with metastatic cancer, pain caused by cancer treatment chemotherapeutic agent, trigeminal neuralgia, burning pain, acute herpes and postherpetic neuralgia, laryngeal neuralgia, sympathetic dystrophy, fibromyalgia, gout pain, burn pain, phantom limb pain, complex regional pain syndrome, idiopathic pain syndrome, migraine, and chronic pain of unknown cause. A composition comprising
The peptide BigLEN having the amino acid sequence of SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof, a diastereomer, an enantiomer, and a peptide selected from the group consisting of amino acid substitution orthologs as an active ingredient for pain suppression Or a functional health food for pain prevention:
<SEQ ID NO: 1>
LENPSPEAPARRRLLPP.
The health functional food according to claim 1, wherein the amino acid constituting the amino acid sequence of SEQ ID NO: 1 is an L-amino acid.
The health functional food according to claim 6, wherein the amino acid constituting the amino acid sequence of SEQ ID NO: 1 is an L-amino acid.
The health functional food according to claim 6, wherein the centromere has one or more amino acid substitutions selected from the group consisting of N3T, S5A, E7Q and A8V in the amino acid sequence of SEQ ID NO: 1.

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