KR102425548B1 - Cacb1-derived peptide, Cacb1-derived peptide variants and uses thereof - Google Patents

Abstract

The present invention relates to a Cacb1-derived peptide and a variant of the Cacb1-derived peptide, and a composition for preventing or treating hypertension and a composition for vasodilation comprising the same. The Cacb1-derived peptide or variant of the Cacb1-derived peptide of the present invention has excellent blood pressure lowering effect, has fewer side effects, and is easy to manufacture and industrialize, so it can be usefully used in the development of new antihypertensive drugs. In addition, due to its potential vasodilatory effect, it is expected to be used as a therapeutic candidate for diseases that can be treated or improved by vasodilation, such as angina pectoris, prostatic hypertrophy, and erectile dysfunction.

Description

Cacb1-derived peptide, Cacb1-derived peptide variant and uses thereof {Cacb1-derived peptide, Cacb1-derived peptide variants and uses thereof}

The present invention relates to a Cacb1-derived peptide, a Cacb1-derived peptide variant, and a composition for preventing or treating hypertension and a composition for vasodilation comprising the same.

Hypertension is a disease with a high prevalence rate, affecting one in three adults in most countries. According to the 2016 National Health and Nutrition Survey, the prevalence of hypertension in Korea is over 11 million. High blood pressure, if left untreated, is often called the “silent killer” because it injures the heart and blood vessels without any symptoms and leads to stroke, acute myocardial infarction, and death through a process called atherosclerosis.

For the treatment of hypertension, drug development has been carried out since the 1950s, and the antihypertensive effect is effective, safety has been proven, and side effects are relatively mild. and alphabeta blockers, calcium antagonists, ACE inhibitors (Angiotensin converting enzyme inhibitors), ARB blockers (Angiotensin receptor blockers). Although these first-choice antidepressants are similar to each other in the strength of the coercive effect, when applied individually to patients, they can show significant differences in morbidity, mortality, and side effects.

On the other hand, the conventionally used compounds are easily decomposed in the pharmaceutical dosage form and not only have poor stability, but also act on other cells to cause body weakness, vomiting, cough, headache, anorexia and taste disorders, etc. It is reported that there is a problem. For example, in the case of thiazide diuretics, it is the oldest antihypertensive agent with well-proven efficacy and safety, and is used as the first-line drug for gout, hyperuricemia, hypokalemia, hyponatremia, hypercalcemia, dyslipidemia, impaired glucose tolerance, It is known to cause erectile dysfunction. In addition, the launch of a new concept of antihypertensive drug for the past 10 years is minimal, and the pharmaceutical market is preparing to launch a combination drug combining two or more drug series among existing antihypertensive drugs.

In the case of high blood pressure, the drug must be taken for a long time due to the nature of the disease, and if the drug is abruptly stopped, greater side effects may appear. Accordingly, there is an urgent need to develop a new antihypertensive agent that can replace the existing antihypertensive drug and can be applied to patients who have not been treated with the existing antihypertensive drug.

1. US Patent Application Publication US2010/0196355 (2010.08.05.) 2. US Patent Application Publication US2015/0250854 (2015.09.10.) 3. Patent Publication No. 10-2015-0145132 (2015.12.29.)

Accordingly, the present inventors made efforts to develop a novel peptide-based antihypertensive agent, confirmed the blood pressure lowering effect of a Cacb1-derived peptide and a variant thereof, and completed the present invention.

Therefore, an object of the present invention is a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) To provide a pharmaceutical composition for preventing or treating hypertension, a composition for vasodilation, a food composition, and a method for preventing or treating hypertension, comprising a fusion peptide fused with a Cacb1-derived peptide; and a cell-permeable peptide or half-life extension peptide.

In order to achieve the above object,

The present invention is a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a pharmaceutical composition for preventing or treating hypertension comprising a fusion peptide.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a composition for vasodilation comprising a fusion peptide.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extension peptide; provides a food composition for preventing or improving hypertension comprising a fusion peptide.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a method for preventing or treating hypertension, comprising administering to a subject a fusion peptide.

The Cacb1-derived peptide and variants of the Cacb1-derived peptide of the present invention have excellent blood pressure lowering effects, have fewer side effects, and are easy to manufacture and industrialize, so they can be usefully used in the development of new antihypertensive drugs. In addition, due to its potential vasodilatory effect, it is expected to be used as a therapeutic candidate for diseases that can be treated or improved by vasodilation, such as angina pectoris, prostatic hypertrophy, and erectile dysfunction.

1 is an observation of changes in blood pressure during intravenous injection of Cacb1 (344-359) peptide (IKITSPKVLQRLIKSR) into a rat animal model, and the control group and Cacb1 (344-359) (0.15 mg/kg; 0.2ml, IV) treatment group Blood pressure comparison (A), Cacb1 (344-359) (1 mg/kg; 0.2ml, IV) treatment group (B, C) is a diagram showing changes in blood pressure.
2 is a diagram showing changes in blood pressure during intravenous injection of Cacb1 (389-397) peptide (LDENQLEDA) into an animal model.
3 is a diagram showing changes in blood pressure upon injection of Cacb1(344-359)I344V peptide (VKITSPKVLQRLIKSR) (A) or Cacb1(344-359)I344V/K357R peptide (VKITSPKVLQRLIRSR) (B) into an animal model.
4 is a diagram showing blood pressure change (A) and blood pressure change when intramuscular injection (B) of Cacb1(344-359)K357R peptide (IKITSPKVLQRLIRSR) intravenously injected into an animal model.
5 is a Cacb1(344-359) peptide, Cacb1(389-397) peptide, Cacb1(344-359)I344V peptide, Cacb1(344-359)I344V/K357R peptide or Cacb1(344-359)K357R peptide in an animal model. Changes in systolic blood pressure during intravenous injection (A), blood pressure (B) and hemodynamic parameters (systolic, diastolic, and mean blood pressure) of each animal model before and after administration of Cacb1 (344-359) peptide (C) ; Numbers in parentheses are the number of animal models used for average calculation).
6 is a diagram showing changes in blood pressure upon injection of Cacb1 (344-359) I344V/T347S peptide (VKISSPKVLQRLIKSR) (A) or Cacb1 (344-357) I344V/T347S/K357R peptide (VKISSPKVLQRLIRSR) (B) into an animal model.
Figure 7 shows the change in blood pressure upon injection of Cacb1(344-357)I344V/I346V/T347S peptide (VKVSSPKVLQRLIKSR) (A) or Cacb1(344-357)I344V/I346V/T347S/K357R peptide (VKVSSPKVLQRLIRSR) (B) into an animal model. is the diagram shown.
Figure 8 shows Cacb1 (344-359), Cacb1 (389-397), Cacb1 (344-359) I344V/T347S (Cacb2 (392-407)), Cacb1 (344-357) I344V/T347S/K357R (Cacb2 (392) -407)K405R), Cacb1(344-357)I344V/I346V/T347S(Cacb4(333-348)) or Cacb1(344-357)I344V/I346V/T347S/K357R(Cacb3(292-307)) systolic blood pressure Changes (A) and hemodynamic parameters (systolic, diastolic, and mean blood pressure) were measured (B; the number in parentheses is the number of animal models used to calculate the average).
9 is a diagram showing the blood pressure change (A) and hemodynamic indices (systolic, diastolic, and mean blood pressure) measured for each concentration when TAT-Cacb1 (389-397) fusion peptide is injected into an animal model (B).
Figure 10 shows blood pressure changes (A) and Cacb1(344-359)K357R, peptide K2-palm peptide (0.5, 1, 2 mg/kg) or Results of hemodynamic parameters (systolic, diastolic, mean blood pressure and heart rate) measured during intravenous infusion of K7-palm peptide (2 mg/kg) (B; the number in parentheses is the number of animal models used for averaging) is a diagram showing
11 shows blood pressure changes (A) and Cacb1(344-359)K357R peptide, Cacb1(344-359)K357R-EYEKEYE peptide (0.5, 1, 2 mg/kg) or K20-palm peptide (2 mg/kg) intravenous infusion of hemodynamic parameters (systolic, diastolic, mean blood pressure and heart rate) measurement results (B; numbers in parentheses are used for average calculation) It is a diagram showing the number of animal models).

Hereinafter, the present invention will be described in detail.

The present invention is a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a pharmaceutical composition for preventing or treating hypertension comprising a fusion peptide.

In the present invention, the “Cacb1” gene is a protein constituting a calcium channel, among others, a gene encoding a calcium channel β1 subunit, and refers to a voltage-dependent L-type calcium channel subunit beta-1 gene, to which the present invention belongs. In the art, the gene is also referred to as CAB1, Cacnb, Cacnlb1, Cchb1, Cchlb1, calcium channel voltage-dependent subunit beta 1 or voltage-dependent calcium channel beta1 subunit.

In an embodiment of the present invention, a peptide derived from Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1, calcium channel β1 subunit) (protein ID P54283.1(rat)) was used in the experiment.

In the present invention, the "Cacb1-derived peptide" is used to include all of the Cacb1-derived peptide, a Cacb1-derived peptide variant, and a fatty acid-peptide complex in which a fatty acid is additionally bound to a Cacb1-derived peptide variant.

In the present invention, the term “fusion” refers to the integration of two molecules with different or the same function or structure, which is any physical, chemical, or biological method capable of binding the cell-permeable peptide or the half-life extension peptide to the Cacb1-derived peptide according to the present invention. fusion by methods is included without limitation.

In the present invention, the term “fusion peptide” refers to a peptide in which the derived peptide and a functional peptide are fused. It may be any one selected from the group consisting of 22, and 70% or more, preferably 80% or more, more preferably 90% of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 11 to 19 and SEQ ID NO: 22 or more, and most preferably, an amino acid sequence having sequence homology of 95% or more.

In the present invention, the term “variant” refers to a peptide in which one or more amino acid residues in the Cacb1-derived peptide are deleted, added, inserted or substituted within the range capable of exerting the blood pressure lowering effect according to the present invention. In the present invention, the Cacb1-derived peptide may consist of the amino acid sequence shown in SEQ ID NO: 1, and the variant of the Cacb1-derived peptide may be one in which one or more amino acid residues are substituted in the amino acid of SEQ ID NO: 1.

The variant of the Cacb1 peptide according to the present invention is the substitution of valine (V) for isoleucine (I) at amino acid 344 in the amino acid of SEQ ID NO: 1, and arginine for lysine (K) at amino acid 357 , R), Threonine (T) at amino acid position 347 with Serine (S), and Isoleucine (I) at amino acid position 346 with Valine (V). It may include one or more substitutions selected from the group, and may preferably be represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs: 2 to 8, and any one selected from the group consisting of SEQ ID NOs: 2 to 8 It may comprise an amino acid sequence having at least 70%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95% sequence homology to the amino acid sequence.

Meanwhile, those skilled in the art may modify some sequence of the peptide according to the present invention or reduce the size thereof, or use a known peptide modification method (terminal modification, cyclic peptide, fatty acid fusion, peptide fusion, etc.) to lower blood pressure It will be obvious that the size, duration, and bioavailability can be improved.

In an experimental example of the present invention, it was confirmed that a significant blood pressure lowering effect was observed when the peptide represented by the amino acid sequence of SEQ ID NOs: 1 to 8 was injected into an animal model.

On the other hand, as a result of sequencing, the amino acid sequence of SEQ ID NO: 1 according to the present invention is identical to the amino acid sequence (SEQ ID NO: 25) of Human β1b (389-404) corresponding to the rat Cacb1 (344-359) amino acid sequence, and SEQ ID NO: 5 The amino acid sequence of is identical to the amino acid sequence (SEQ ID NO: 26) of Human β2a (341-356) or Human β2b (342-357) corresponding to the rat Cacb2 (392-407) amino acid sequence, and the amino acid sequence of SEQ ID NO: 8 is It is identical to the amino acid sequence (SEQ ID NO: 27) of Human β3 (292-307) corresponding to Rat Cacb3 (292-307), and the amino acid sequence of SEQ ID NO: 7 is Human β4 (287) corresponding to Rat Cacb4 (333-348) -302) was confirmed to be identical to the amino acid sequence (SEQ ID NO: 28). Therefore, it is obvious that those skilled in the art can use, without limitation, a peptide derived from the protein Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1, calcium channel β1 subunit) constituting the rat or human calcium channel as the peptide according to the present invention. something to do.

The "fatty acid-peptide complex" of the present invention may be a form in which a fatty acid is additionally bound to a Cacb1-derived peptide variant, and in the present invention, the Cacb1-derived peptide variant is represented by any one selected from the group consisting of SEQ ID NOs: 1 to 8 It may be an amino acid sequence, more preferably the amino acid sequence represented by SEQ ID NO: 4, and 70% or more, preferably 80% or more, more preferably 90% or more, most preferably the amino acid sequence represented by SEQ ID NO: 4 Preferably, it may include an amino acid sequence having at least 95% sequence homology. In addition, in the present invention, the fatty acid is myristic acid, stearic acid, linoleic acid, palmitic acid, oleic acid, and lauric acid. (Lauric acid) may be at least one selected from the group consisting of, more preferably palmitic acid (Palmitic acid). The fatty acid may be bound to any position of the Cacb1-derived peptide mutant peptide, preferably to lysine among amino acids. In one embodiment, the fatty acid-peptide complex according to the present invention may be any one selected from the group consisting of SEQ ID NOs: 20 and 21, and 70% or more with any one amino acid sequence selected from the group consisting of SEQ ID NOs: 20 and 21 , preferably 80% or more, more preferably 90% or more, and most preferably 95% or more of an amino acid sequence having sequence homology.

In the present invention, the term “Cell Penetrating Peptides (CPP)” refers to about 10-30 short cell membrane-penetrating peptides, mostly protein-transduction domains (PTDs) or membrane-transfer sequences (Membrane). -Translocating Sequence (MTS) is also called PTD or MTS. The cell-penetrating peptide can efficiently transport the fusion protein into a cell even when it is linked to another peptide or protein. In the present invention, the cell-permeable peptide is a known cell-penetrating peptide capable of penetrating a cell membrane, for example, pep-1 peptide (KETWWETWWTEWSQPKKKRKV), low molecular weight protamine (LMWP, VSRRRRRRGGRRRR), TAT Peptide (TAT peptide, YGRKKRRQRRR), Penetratin (RQIKIWFQNRRMKWKK), Antennapedia (ANTP), D-arginine oligopeptide (R8 or higher) or L-arginine oligopeptide (L-arginine oligopeptide) , R8 or more), but is not limited thereto, and may preferably be a TAT peptide represented by the amino acid sequence of SEQ ID NO: 10.

In the present invention, the term 'half-life extending peptide' is used to include both half-life extending peptides and half-life extending peptides and peptides in which fatty acids are bound, and more preferably, half-life extending peptides may be peptides in which fatty acids are bound. The half-life extending peptide is a peptide having a characteristic that can extend the half-life when linked to other peptides and proteins, and may be a peptide consisting of about 2 to 12 amino acids, preferably a peptide consisting of 5 to 10 amino acids. More preferably, it may be a peptide consisting of 7 peptides, and more preferably it may be a peptide consisting of the amino acid sequence shown in SEQ ID NO: 24, and 70% or more, preferably 80%, of the amino acid sequence shown in SEQ ID NO: 24 It may include an amino acid sequence having at least 90% sequence homology, more preferably at least 90% sequence homology, and most preferably at least 95% sequence homology.

In addition, a fatty acid may be additionally bound to the half-life extending peptide, and the fatty acid is myristic acid, stearic acid, linoleic acid, palmitic acid, oleic acid ( Oleic acid) and lauric acid may be at least one selected from the group consisting of, more preferably palmitic acid. The peptide to which a fatty acid is bound to the half-life extension peptide may be a peptide consisting of the amino acid sequence represented by SEQ ID NO: 23, and 70% or more, preferably 80% or more, more preferably 90% or more of the amino acid sequence represented by SEQ ID NO: 23 , most preferably an amino acid sequence having at least 95% sequence homology. The fatty acid may be bound to any position among amino acids of the half-life extending peptide, and preferably may be bound to lysine among amino acids of the half-life extending peptide, but is not limited thereto.

In the present invention, the Cacb1-derived peptide may be a peptide represented by the amino acid sequence of SEQ ID NO: 1, a variant thereof, or a peptide represented by the amino acid sequence of SEQ ID NO: 9, wherein the variant is amino acid 344 from the amino acid of SEQ ID NO: 1 Substitution of phosphorus isoleucine (I) with valine (V), lysine (K) at amino acid 357 with arginine (R), substitution of amino acid at 347 with Threonine (T) It may include one or more substitutions selected from the group consisting of substitution with serine (S) and substitution of valine (V) for isoleucine (I), which is amino acid 346, and preferably SEQ ID NO: 2 to any one amino acid sequence selected from the group consisting of 8, and 70% or more, preferably 80% or more, with any one amino acid sequence selected from the group consisting of SEQ ID NOs: 2 to 8, more preferably It may comprise an amino acid sequence having at least 90%, most preferably at least 95% sequence homology.

In the present invention, the fusion peptide is a fusion of a Cacb1-derived peptide according to the present invention and a cell-permeable peptide or a half-life extension peptide, preferably represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs: 11 to 23 it could be

On the other hand, in an experimental example of the present invention, it was confirmed that a significant blood pressure lowering effect was exhibited when the cell-permeable peptide was fused to a Cacb1-derived peptide, which had no blood pressure lowering effect when administered alone. It is possible to increase the blood pressure lowering effect of the derived peptide.

In addition, in an experimental example of the present invention, it was confirmed that the duration of the blood pressure lowering effect significantly increased when a fatty acid was combined with a Cacb1-derived peptide or a half-life extension peptide was fused to a Cacb1-derived peptide, which exhibited a blood pressure lowering effect of about 5 minutes when administered alone. Fatty acids or half-life extending peptides can increase the blood pressure drop duration of Cacb1-derived peptides.

The pharmaceutical composition comprising the peptide according to the present invention as an active ingredient is not limited thereto, but may include one or more of a pharmaceutical diluent selected from saline, buffered saline, dextrose, water, glycerol, and ethanol.

The composition according to the present invention may be applied differently depending on the purpose of administration and disease. The amount of the active ingredient actually administered depends on various related factors, i.e., the disease to be treated, the degree of the patient's condition, whether or not co-administered with other drugs, the patient's age and sex, weight, food, administration time, administration route, and administration ratio of the composition (ratio) should be taken into consideration. The dosage and route of administration of the composition may be adjusted according to the type and severity of the disease, and may be administered once a day or divided into 1 to 3 times, but is not limited thereto.

The composition according to the present invention may be administered orally or parenterally. Parenteral administration refers to administration of drugs via routes other than oral administration, ie, rectal, intravenous, peritoneal and muscle, arterial, transdermal, nasal, inhalational, ocular, and subcutaneous.

The composition may be formulated in any form, such as an oral dosage form, an injectable solution, or a topical preparation. The formulation is preferably prepared for oral and injectable administration (true solution, suspension or emulsion), and is most preferably prepared in oral form such as tablets, capsules, soft capsules, aqueous pharmaceuticals, pills, granules, etc. desirable.

In the above formulation, the peptide of the present invention may be filled into a soft capsule without an excipient, and may be mixed or diluted with a carrier to prepare a suitable formulation. Examples of suitable carriers include starch, water, saline, Ringer's solution, dextrose and the like.

The composition can be directly applied to animals, including humans. The animal is a biological group corresponding to the plant, mainly ingesting organic matter as nutrients, and is said to have differentiated digestion, excretion and respiratory organs, preferably a vertebrate, and more preferably a mammal. The mammal may be a human, a dog, a cat, a mouse, a pig, a cow, or a goat, and preferably a human.

The pharmaceutical composition according to the present invention may include, as an active ingredient, a peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23, and other formulations, methods of use, and purposes of use Additional ingredients, that is, pharmaceutically acceptable or nutritionally acceptable carriers, excipients, diluents or auxiliary ingredients may be further included.

In more detail, the pharmaceutical composition is in addition to the active ingredients, nutrients, vitamins, electrolytes, flavoring agents, coloring agents, thickeners, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, It may further contain stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. In addition, the carrier, excipient or diluent is lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose Rose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, physiological saline It may be one or more, but is not limited thereto, and all of the usual carriers, excipients or diluents may be used. The components may be added independently or in combination to the active ingredient, the peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23.

The pharmaceutical composition contains 0.001% to 99.9% by weight, preferably 0.1% to 99% by weight of a peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23, based on the total weight of the composition , more preferably 1% to 50% by weight.

In addition, the pharmaceutical composition may further include conventional fillers, extenders, binders, disintegrants, surfactants, anti-aggregants, lubricants, wetting agents, fragrances, emulsifiers or preservatives, etc., when medicated, oral or parenteral all can be used

Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, water It may be prepared by mixing sucrose or lactose, gelatin, or the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.

In addition, the formulation of the pharmaceutical composition according to the present invention may be in a desired form depending on the method of use, and in particular, methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal It can be adopted and formulated. Examples of specific formulations include warning agents, granules, lotions, liniment agents, limonade agents, powders, syrups, eye ointments, liquids, aerosols, EXTRACTS, elixirs, ointments, fluid extracts, emulsions, suspensions, preparations, infusions, eye drops, tablets, suppositories, injections, spirits, capsules, creams, pills, soft or hard gelatin capsules, and the like.

The dosage of the pharmaceutical composition according to the present invention may be appropriately selected by those skilled in the art in consideration of the administration method, the age, sex and weight of the user, and the severity of the disease. For example, the pharmaceutical composition of the present invention is based on a peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23, 0.000001 mg/kg/day to 1000 mg/kg/ It can be administered daily. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.

In addition, the pharmaceutical composition according to the present invention is a compound having a known antihypertensive effect in addition to the peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23, which are active ingredients, particularly used as food. It may further include a material derived from a natural product, and may be included in an amount of 5 to 100 parts by weight, respectively, based on 100 parts by weight of the active ingredient.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a composition for vasodilation comprising a fusion peptide.

The composition may be provided in the form of a pharmaceutical composition.

As the composition according to the present invention exerts a vasodilatory or dilating effect, it may ultimately be used for the prevention or treatment of diseases caused by vasodilatory disorders or diseases that can be treated or ameliorated by vasodilation, and the diseases include Hypertensive complications, heart failure, atherosclerosis, angina pectoris, myocardial infarction, cerebral infarction, stroke, cerebral hemorrhage, cardiovascular diseases including peripheral blood circulation disorders, enlarged prostate or erectile dysfunction, and the like, but are not limited thereto. According to a recent study, it has been reported that erectile dysfunction and enlarged prostate are not a single disease but are closely related to other vascular diseases, especially cardiovascular diseases, and vasodilation disorders or structural abnormalities of blood vessels are found in these diseases, Many erectile dysfunction patients also have various vascular diseases such as high blood pressure and peripheral arterial disease. According to one embodiment of the present invention, the Cacb1-derived peptide or the cell-permeable peptide or the fusion peptide in which the half-life extension peptide and the Cacb1-derived peptide are fused according to the present invention shows a remarkable blood pressure lowering effect, and its potential vasodilatory or dilating effect It will be apparent to those skilled in the art that it can ultimately be used for the prevention or treatment of cardiovascular diseases, enlarged prostate, or erectile dysfunction.

In the present invention, the term "prevention" refers to any action of inhibiting or delaying the progression of a disease that can be treated or ameliorated by vasodilation, including hypertension, by administration of the pharmaceutical composition according to the present invention.

In the present invention, the term "treatment" refers to any action in which a disease that can be treated or ameliorated by vasodilation, including hypertension, is ameliorated or changed advantageously by administration of the pharmaceutical composition according to the present invention.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extension peptide; provides a food composition for preventing or improving hypertension comprising a fusion peptide.

The type of the food is not particularly limited, but is preferably a health functional food. Examples of foods to which the above substances can be added include various foods, beverages, gum, candy, tea, vitamin complexes, functional foods, etc., which include special nutritional foods (eg, formula milk, infant food, etc.), processed meat products, Fish meat products, tofu, jelly, noodles (eg, ramen, noodles, etc.), health supplements, seasonings (eg, soy sauce, soybean paste, red pepper paste, mixed soy sauce, etc.), sauces, confectionery (eg snacks), dairy products (eg, Fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi, pickles, etc.), beverages (eg fruit, vegetable beverages, soy milk, fermented beverages, ice cream, etc.), natural seasonings (eg ramen soup, etc.) ), vitamin complexes, alcoholic beverages, alcoholic beverages and other health supplements, but are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.

The peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23 of the present invention can be added to food as it is or used with other foods or food ingredients, and can be used appropriately according to a conventional method. have. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of its use (for prevention or improvement). In general, the amount of the peptide in the health food may be added in an amount of 0.01 to 15% by weight, preferably 0.1 to 5% by weight of the total food weight, and 0.01 to 5.0 g, preferably 0.01 to 5.0 g based on 100 in the health beverage composition. It can be added in a proportion of 1.0 g. However, in the case of long-term ingestion for the purpose of health control, the above amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

The health functional beverage composition according to the present invention is not particularly limited in other ingredients except for containing a peptide as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; conventional sugars such as disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, erythritol and the like. As flavoring agents other than those described above, natural flavoring agents (taumakin, stevia extract, etc.), and synthetic flavoring agents (saccharin, aspartan, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 0.1 to 2.0 g, preferably about 0.1 to 1.0 g per 100 compositions of the present invention.

The health functional food of the present invention can be prepared by adding the antibiotic peptide consisting of the amino acid sequence of the present invention described above during the manufacturing process of the base food or after preparing the base food, the amino acid sequence of the present invention. It can be easily obtained by adding a step of adding the constituting peptide. At this time, if necessary, a taste and odor correcting agent may be added.

In a food composition for the purpose of preventing or improving hypertension, the amount of the peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23 is 0.00001% to 50% by weight of the total food weight. may be included, but is not limited thereto.

In addition, the present invention a) Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide; or

b) Cacb1-derived peptide; and cell-permeable peptide or half-life extending peptide; provides a method for preventing or treating hypertension, comprising administering to a subject a fusion peptide.

The subject may be a mammal including a human. The subject may be suffering from, or likely to suffer from, hypertension.

The peptide consisting of any one amino acid sequence selected from SEQ ID NOs: 1 to 8 and SEQ ID NOs: 11 to 23 of the present invention may be administered to the subject, and the administration method may be oral or parenteral administration. The method of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, topical, intranasal, intratracheal, or intradermal routes, but includes It is not limited.

In the present specification, terms and definitions used in the composition for vasodilation, the food composition for preventing or improving hypertension, and the method for preventing or treating hypertension are the same as the pharmaceutical composition for preventing or treating hypertension, unless otherwise specified, in consideration of overlap and complexity. Description is omitted.

Terms not otherwise defined herein have the meanings commonly used in the art to which the present invention pertains.

Hereinafter, the present invention will be described in detail by way of Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Example 1. Preparation of Cacb1 variants according to the present invention

A peptide derived from the protein Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1, calcium channel β1 subunit) constituting the calcium channel (protein ID P54283.1(rat)), specifically, amino acid sequence 344 of the Cacb1 protein Peptide IKITSPKVLQRLIKSR (hereinafter, referred to as “Cacb1 (344-359)”) consisting of Nos. to 359 (rat basis) and variants thereof were used in the experiment. All peptides were purchased from Peptron Co., Ltd. (Daejeon, Korea) and the synthesis was made through solid-phase peptide synthesis by Fmoc chemistry. Specifically, each amino acid was synthesized using an automatic synthesizer by synthesizing each amino acid one by one in resin according to a predetermined sequence, and the synthesized crude peptide was purified using reverse-phase HPLC and dried with a freeze dryer. was used. The molecular weight of Cacb1 (344-359) is 1879 Da, and it has relatively good solubility in water (solubility in water 10 mg/ml), so it was dissolved in 0.9% NaCl (saline, Saline) and used in the experiment.

The amino acid sequences of all the peptides used in the present invention are shown in Table 1 below.

name order SEQ ID NO: Cacb1 (344-359) IKITSPKVLQRLIKSR One Cacb1(344-359) I344V V KITSPKVLQRLIKSR 2 Cacb1(344-359) I344V/K357R V KITSPKVLQRLI R SR 3 Cacb1 (344-359) K357R IKITSPKVLQRLI R SR 4 ^* Cacb1(344-359) I344V/T347S V KI S SPKVLQRLIKSR 5 Cacb1(344-359) I344V/T347S/K357R V KI S SPKVLQRLI R SR 6 ^# Cacb1(344-359) I344V/I346V/T347S V K VS SPKVLQRLIKSR 7 ^$ Cacb1(344-359) I344V/I346V/T347S/K357R V K VS SPKVLQRLI R SR 8 Cacb1 (389-397) LDENQLEDA 9 TAT YGRKKRRQRRR 10 TAT-Cacb1 (344-359) YGRKKRRQRRR-IKITSPKVLQRLIKSR 11 TAT-Cacb1(344-359) I344V YGRKKRRQRRR- V KITSPKVLQRLIKSR 12 TAT-Cacb1(344-359) I344V/K357R YGRKKRRQRRR- V KITSPKVLQRLI R SR 13 TAT-Cacb1 (344-359) K357R YGRKKRRQRRR-IKITSPKVLQRLI R SR 14 TAT- ^* Cacb1 (344-359) I344V/T347S YGRKKRRQRRR- V KI S SPKVLQRLIKSR 15 TAT-Cacb1(344-359) I344V/T347S/K357R YGRKKRRQRRR- V KI S SPKVLQRLI R SR 16 TAT- ^# Cacb1(344-359) I344V/I346V/T347S YGRKKRRQRRR- V K VS SPKVLQRLIKSR 17 TAT- ^$ Cacb1(344-359) I344V/I346V/T347S/K357R YGRKKRRQRRR- V K VS SPKVLQRLI R SR 18 TAT-Cacb1 (389-397) YGRKKRRQRRR-LDENQLEDA 19 K2-palm IK (palmitoyl )ITSPKVLQRLIRSR 20 K7-palm IKITSPK (palmitoyl )VLQRLIRSR 21 Cacb1(344-359)K357R-EYEKEYE IKITSPKVLQRLIRSR- EYEKEYE 22 K20-palm IKITSPKVLQRLIRSR- EYEK(palmitoyl)EYE 23 EYEKEYE EYEKEYE 24

*Cacb1 (344-359) I344V/T347S (SEQ ID NO: 5) is identical to the amino acid sequence of the existing Cacb2 (392-407) [protein ID Q8VGC3.2 (rat)]. #Cacb1(344-359) I344V/I346V/T347S (SEQ ID NO: 7) is identical to the amino acid sequence of the existing Cacb4(333-348) [protein ID D4A055.2(rat)]. $Cacb1 (344-359) I344V / I346V / T347S / K357R (SEQ ID NO: 8) is identical to the amino acid sequence of the existing Cacb3 (292-307) [protein ID P54287.1 (rat)].

In addition, the peptide sequence derived from human Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1, calcium channel β1 subunit) (protein ID: AAB58781.1 (human)) corresponding to the rat reference amino acid sequence of Table 1 above. It is shown in Table 2 below.

name ^* Sequence SEQ ID NO: Human β1b (389-404) (corresponding to Rat Cacb1 (344-359)) IKITSPKVLQRLIKSR 25 Human β2a (341-356) or Human β2b (342-357)
(corresponds to Rat Cacb2 (392-407)) VKISSPKVLQRLIKSR 26 Human β3 (292-307) (corresponding to Rat Cacb3 (292-307)) VKVSSPKVLQRLIRSR 27 Human β4 (287-302) (corresponding to Rat Cacb4 (333-348)) VKVSSPKVLQRLIKSR 28

*SEQ ID NO: 25 is identical to the amino acid sequence of Cacb1 (344-359) (SEQ ID NO: 1); SEQ ID NO: 26 is identical to the amino acid sequence of Cacb1 (344-359) I344V/T347S (SEQ ID NO: 5); SEQ ID NO: 27 is identical to the amino acid sequence of Cacb1 (344-359) I344V/I346V/T347S/K357R (SEQ ID NO: 8); SEQ ID NO: 28 is identical to the amino acid sequence of Cacb1 (344-359) I344V/I346V/T347S (SEQ ID NO: 7).

Example 2. Femoral artery and vein catheterization in a rat animal model

All experiments were conducted with the approval of the Experimental Animal Ethics Committee of Kyungpook National University. Male and female rats (Sprague-Dawley rats) weighing about 330 to 430 g were used as experimental animals, and a catheter was inserted into the femoral artery and vein to measure arterial blood pressure and inject the peptide solution according to the present invention. .

First, the rat is anesthetized by supplying isoflurane (2%) along with oxygen (1-1.5%), intraperitoneal urethane (1g/kg) injection and isoflurane (0.8- 1%) by injection to maintain anesthesia. The femoral skin was incised to expose the femoral nervous system bundle, and the femoral sheath was removed by making an incision along the long axis between the femoral artery and vein. Then, the femoral artery is pulled laterally and the femoral vein is pulled inward, and two silk sutures (4-0 silk) are inserted under the artery located just below the arteria profunda femoris, and one is loosely ligated. Then, it was pulled toward the body, and the other was tightly ligated at the point farthest possible toward the leg. Similarly, two silk sutures (4-0 silk) were inserted under the vein located below the deep femoral vein (vena profunda femoris), and the silk thread was loosely ligated for subsequent proximal and distal ligation. Then, a micro incision was made about 2 mm at the distal ligation site of each femoral artery or femoral vein, and a 1 mL syringe pre-filled with saline solution containing 300 U/L heparin (catheter, PE50 tube) was inserted. When the catheter was initially inserted, the proximal ligation was loosened and the catheter was fully inserted (>1 cm), and then the proximal ligation was finished with a triple knot around the artery and catheter or around the vein and the catheter to fix the catheter to the blood vessel. Blood inside the inserted catheter was checked, and catheter patency was confirmed by flushing heparinized saline solution.

Example 3. Measurement of Arterial Blood Pressure in Experimental Animals

Example 3. Measurement of Arterial Blood Pressure in Experimental Animals

The catheter inserted through the femoral artery was connected to a PowerLab Data acquisition system (ADInstruments, Dunedin, Zew Zealand) and connected to a pressure transducer placed flush with the heart of the rat. Arterial blood pressure was analyzed by LabChart (version 7, AD Instruments). The systolic pressure was identified as the peak, and the diastolic pressure was identified as the valley between the maximum value and the maximum value, and the mean arterial pressure was the diastolic blood pressure and 1/ the difference between the systolic and diastolic blood pressures. 3 was added to calculate. Heart rate was determined as the number of cycles per minute. In the figure, the change in blood pressure (Δ) was calculated by subtracting the blood pressure after injection from the blood pressure before the injection of the peptide solution. Statistical analysis was performed by Student T-test (*p < 0.05, **p < 0.01), and analysis values were expressed as mean ± standard error (SEM). The results are shown in Table 3 below.

Experimental Example 1. Confirmation of blood pressure lowering effect of Cacb1-derived peptide

In order to select a peptide candidate that induces a decrease in blood pressure when administered to an animal model, the blood pressure lowering effect of two peptides having a length of 9 or 16 amino acids derived from the Cacb1 protein was confirmed.

1-1. Confirmation of blood pressure lowering effect of Cacb1(344-359) peptide

Fig. 1 shows the results of dissolving the Cacb1 (344-359) peptide (IKITSPKVLQRLIKSR, SEQ ID NO: 1) prepared in Example 1 in 0.9% NaCl and injecting it into the femoral vein of an adult rat. As shown in FIG. 1A , it was confirmed that the systolic and diastolic blood pressure recorded from the femoral artery was significantly reduced at a dose of 0.15 mg/kg or more. In addition, as shown in FIG. 1B , it was confirmed that a larger and more persistent blood pressure lowering effect appeared when the dose was treated at 1 mg/kg, and as shown in FIG. 1C , the blood pressure change was analyzed in seconds. As a result, it was confirmed that the systolic and diastolic blood pressure decreased (2, about 60/30 mmHg) after administration (2, about 60/30 mmHg) than before (1, about 130/90 mmHg) administration of the Cacb (344-359) peptide, and then recovered (3, about 130). /90 mmHg).

1-2. Confirmation of blood pressure lowering effect of Cacb(389-397) peptide

The result of dissolving the peptide LDENQLEDA (hereinafter referred to as “Cacb1 (389-397)”) (SEQ ID NO: 9) consisting of amino acid sequences 389 to 397 (rat basis) derived from the same Cacb1 protein in physiological saline and administering it to the femoral vein is shown in FIG. 2 . As shown in FIG. 2 , it was confirmed that the Cacb (389-397) peptide did not affect blood pressure at a dose of 0.5 mg/kg.

Through the above results, it was confirmed that the blood pressure lowered in a concentration-dependent manner according to the administration of the Cacb1(344-359) peptide among the Cacb1-derived peptides. wanted to

Experimental Example 2. Confirmation of the blood pressure lowering effect of the Cacb1 peptide mutant according to the present invention 2-1. Effect analysis of Cacb1(344-359)I344V, Cacb1(344-359)I344V/K357R, Cacb1(344-359)K357R

Peptide VKITSPKVLQRLIKSR (hereinafter, “Cacb1(344-359)I344V”) in which isoleucine (I), amino acid 344, was substituted with valine (V) in Cacb1 (344-359) peptide, which was confirmed to have a blood pressure lowering effect in Experimental Example 1 ”) (SEQ ID NO: 2), the peptide VKITSPKVLQRLIRSR (hereinafter “Cacb1 (344-359) I344V/K357R”) in which lysine (lysine, K), which is amino acid 357, is substituted with arginine (R) ( SEQ ID NO: 3), the blood pressure lowering effect of the peptide IKITSPKVLQRLIRSR (“Cacb1(344-359)K357R”) (SEQ ID NO: 4) in which only the 357 amino acid lysine (K) is substituted with arginine (R), and the results are shown 3 and 4 are shown. In particular, an experiment of injecting intramuscularly other than intravenous injection was additionally performed and the results were confirmed.

As shown in A and B of FIG. 3, Cacb1(344-359)I344V, Cacb1(344-359)I344V/K357R was dissolved in physiological saline and administered by femoral vein at a dose of 0.3 to 0.5 mg/kg or more. It was confirmed that it exhibits a blood pressure lowering effect.

As shown in FIG. 4A , it was confirmed that Cacb1(344-359)K357R also exhibited a significant blood pressure lowering effect. In particular, it was confirmed that Cacb1(344-359)K357R exhibited a drop in arterial blood pressure at a dose of 1.8 mg/kg even when injected into the femoral muscle of a rat (Intramuscular injection) (FIG. 4B).

Experimental results up to Experimental Example 1, Experimental Example 2, and 2-1 are summarized in FIG. 5 . As shown in A and B of FIG. 5 , it was confirmed that Cacb1 (344-359) and its mutant peptides except for the Cacb (389-397) peptide showed a significant blood pressure lowering effect, and as shown in FIG. 5C, 0.5 As a result of measuring hemodynamic parameters when administered at a dose of mg/k, significant differences between the Cacb1 (389-397) peptide and Cacb1 (344-359) and its mutant peptides were found in systolic blood pressure, diastolic blood pressure, and mean arterial pressure. Confirmed.

2-2. Effect analysis of Cacb1(344-359)I344V/T347S, Cacb1(344-357)I344V/T347S/K357R

Peptide VKISSPKVLQRLIKSR in which isoleucine (I) at No. 344 was substituted with valine (V) and threonine at No. 347 with serine (S) in Cacb1 (344-359) peptide whose blood pressure lowering effect was confirmed in Experimental Example 1 (“Cacb1(344-359)I344V/T347S” or “Cacb2(392-407)”) (SEQ ID NO: 5) and the peptide VKISSPKVLQRLIRSR (“Cacb1( The blood pressure lowering effect of 344-357) I344V/T347S/K357R” or “Cacb2(392-407)K405R) (SEQ ID NO: 6) was confirmed, and the results are shown in FIG. 6 .

As shown in FIG. 6A, the Cacb1(344-359)I344V/T347S (or Cacb2(392-407)) peptide showed changes in arterial blood pressure at a dose of 0.5 mg/kg when administered into the femoral vein in an animal model. However, it was confirmed that a significant drop in blood pressure was induced when administered at 0.77 mg/kg. In the case of the Cacb1(344-357)I344V/T347S/K357R peptide, as shown in FIG. 6B , it was confirmed that a significant blood pressure lowering effect was exhibited even at a dose of 0.5 mg/kg.

2-3. Effect analysis of Cacb1(344-357)I344V/I346V/T347S, Cacb1(344-357)I344V/I346V/T347S/K357R

In the Cacb1 (344-359) peptide, which confirmed the blood pressure lowering effect in Experimental Example 1, isoleucines (I) at Nos. 344 and 346 were replaced with valine (V), and Threonine (T) at No. 347 was replaced with Serine (S). peptide VKVSSPKVLQRLIKSR (“Cacb1(344-357)I344V/I346V/T347S” or “cacb4(333-348)”) (SEQ ID NO: 7) and a peptide in which lysine (K) at position 357 is substituted with arginine (R) The blood pressure lowering effect of VKVSSPKVLQRLIRSR (“Cacb1(344-357)I344V/I346V/T347S/K357R” or “Cacb3(292-307)”) (SEQ ID NO: 8) was confirmed, and the results are shown in FIG. 7 .

7, it was confirmed that both Cacb1(344-359)I344V/T347S and Cacb1(344-357)I344V/I346V/T347S/K357R exhibit a significant blood pressure lowering effect when administered into the femoral vein of an animal model.

From the above results, it was confirmed that all of the Cacb1 peptides and variants thereof according to the present invention prepared in Example 1, except for Cacb1 (389-397), could exhibit a clear blood pressure lowering effect. This is summarized in FIG. 8 .

Experimental Example 3. Confirmation of the blood pressure lowering effect of the peptide linking the TAT sequence to Cacb1 (389-397)

The cell-permeable peptide TAT sequence (YGRKKRRQRRR) (SEQ ID NO: 10) was added to the amino acid sequence of the Cacb1 (389-397) peptide, which was confirmed to have no blood pressure lowering effect in Experimental Example 1-2 (hereinafter, “TAT-Cacb ( 389-397)) (SEQ ID NO: 19), the blood pressure lowering effect of the YGRKKRRQRRR-LDENQLEDA peptide was confirmed. The results are shown in FIG. 9 .

As shown in FIG. 9, as a result of administering TAT-Cacb (389-397) into the femoral vein of an animal model, unlike the case of administering Cacb (389-397) alone, it showed a significant concentration-dependent blood pressure lowering effect. Confirmed.

Experimental Example 4. Confirmation of blood pressure lowering effect and duration of peptides added with palmitic acid to Cacb1(344-359)K357R

Coupling palmitic acid to lysine located at the second position among the amino acid sequence of the Cacb1(344-359)K357R peptide, which confirmed the blood pressure lowering effect in Experimental Example 2-1 (hereinafter referred to as “K2-palm”) (SEQ ID NO: 20) Alternatively, palmitic acid was bound (hereinafter, referred to as “K7-palm”) to lysine located at the seventh position (SEQ ID NO: 21), and the blood pressure lowering effect and duration of the peptides were confirmed. The effect was compared with Cacb1 (344-359) K357R (0.5 mg/kg) and the results are shown in FIG. 10 .

As shown in FIG. 10 , when K2-palm was administered into the femoral vein of an animal model, a significant blood pressure lowering effect was induced at a dose of 1 mg/kg, and it was confirmed that the effect lasted more than 2 hours. In addition, when K7-palm was administered at a dose of 2 mg/kg, a blood pressure lowering effect similar to that of K357R was obtained, and it was confirmed that the effect lasted for more than 1 hour. These results show that adding palmitic acid can increase the duration of the blood pressure lowering effect.

Experimental Example 5. Confirmation of blood pressure lowering effect and duration of peptide fused with EYEKEYE peptide to Cacb1(344-359)K357R

Addition of the half-life extension peptide EYEKEYE sequence (SEQ ID NO: 24) to the amino acid sequence of the Cacb1(344-359)K357R peptide, which confirmed the blood pressure lowering effect in Experimental Example 2-1 (hereinafter, “Cacb1(244-259)K357R-EYEKEYE” (SEQ ID NO: 22) or EYEK (pamitoyl) EYE sequence, which is a half-life extension peptide bound to fatty acids (hereinafter referred to as “K20-palm”) (SEQ ID NO: 23), and the blood pressure lowering effect and duration of the peptides was confirmed.

As shown in FIG. 11 , when Cacb1(344-359)K357R-EYEKEYE was administered into the femoral vein of an animal model, a blood pressure lowering effect similar to that of K357R was shown at a dose of 2 mg/kg. In addition, when K20-palm was administered into the femoral vein in an animal model, a significant blood pressure lowering effect was induced at a dose of 2 mg/kg, and it was confirmed that the effect lasted for 1 to 2 hours. These results show that adding a fatty acid-bound half-life extending peptide can increase the duration of the blood pressure lowering effect.

Through the above results, the Cacb1 peptide and the variant of the Cacb1 peptide according to the present invention have excellent blood pressure lowering effect, have biocompatibility and high specificity due to the nature of the peptide, and thus exhibit excellent pharmacological effects while having fewer side effects compared to other antihypertensive drugs. It was confirmed that it can be usefully used in the development of new antihypertensive drugs. In addition, it is expected to be used as a therapeutic candidate for diseases that can be treated or improved by vasodilation, such as angina pectoris, prostatic hypertrophy, and erectile dysfunction due to its potential vasodilatory or dilating effect.

Although the present invention has been described as the above-mentioned preferred embodiment, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also intended that the appended claims cover such modifications and variations as fall within the scope of the present invention.

SEQUENCE LISTING <110> Kyungpook National University Industry-Academic Cooperation Foundation <120> Cacb1-derived peptide, Cacb1-derived peptide variants and uses it <130> KNU1.52PCT <150> KR1020190059969 <151> 2019-05-22 <160> 28 <170> PatentIn version 3.2 <210> 1 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1 (344-359) <400> 1 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 2 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V <400> 2 Val Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 3 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V/K357R <400> 3 Val Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 4 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)K357R <400> 4 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 5 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V/T347S <400> 5 Val Lys Ile Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 6 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V/T347S/K357R <400> 6 Val Lys Ile Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 7 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V/I346V/T347S <400> 7 Val Lys Val Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 8 <211> 16 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)I344V/I346V/T347S/K357R <400> 8 Val Lys Val Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 9 <211> 9 <212> PRT <213> artificial sequence <220> <223> Cacb1 (389-397) <400> 9 Leu Asp Glu Asn Gln Leu Glu Asp Ala 1 5 <210> 10 <211> 11 <212> PRT <213> artificial sequence <220> <223> TAT <400> 10 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg 1 5 10 <210> 11 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1 (344-359) <400> 11 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Ile Lys Ile Thr Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 20 25 <210> 12 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V <400> 12 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Ile Thr Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 20 25 <210> 13 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V/K357R <400> 13 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Ile Thr Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 20 25 <210> 14 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)K357R <400> 14 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Ile Lys Ile Thr Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 20 25 <210> 15 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V/T347S <400> 15 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Ile Ser Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 20 25 <210> 16 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V/T347S/K357R <400> 16 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Ile Ser Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 20 25 <210> 17 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V/I346V/T347S <400> 17 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Val Ser Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 20 25 <210> 18 <211> 27 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1(344-359)I344V/I346V/T347S/K357R <400> 18 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Val Lys Val Ser Ser 1 5 10 15 Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 20 25 <210> 19 <211> 20 <212> PRT <213> artificial sequence <220> <223> TAT-Cacb1 (389-397) <400> 19 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Leu Asp Glu Asn Gln 1 5 10 15 Leu Glu Asp Ala 20 <210> 20 <211> 16 <212> PRT <213> artificial sequence <220> <223> K2-palm <220> <221> MOD_RES <222> (2)..(2) <223> lysine amino acid bound a -palmitoyl group <400> 20 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 21 <211> 16 <212> PRT <213> artificial sequence <220> <223> K7-palm <220> <221> MOD_RES <222> (7)..(7) <223> lysine amino acid bound a -palmitoyl group <400> 21 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 22 <211> 23 <212> PRT <213> artificial sequence <220> <223> Cacb1(344-359)K357R-EYEKEYE <400> 22 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 Glu Tyr Glu Lys Glu Tyr Glu 20 <210> 23 <211> 23 <212> PRT <213> artificial sequence <220> <223> K20-palm <220> <221> MOD_RES <222> (20)..(20) <223> lysine amino acid bound a -palmitoyl group <400> 23 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 Glu Tyr Glu Lys Glu Tyr Glu 20 <210> 24 <211> 7 <212> PRT <213> artificial sequence <220> <223> EYEKEYE <400> 24 Glu Tyr Glu Lys Glu Tyr Glu 1 5 <210> 25 <211> 16 <212> PRT <213> artificial sequence <220> <223> Human β1b (389-404) <400> 25 Ile Lys Ile Thr Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 26 <211> 16 <212> PRT <213> artificial sequence <220> <223> Human β2a (341-356) or Human β2b (342-357) <400> 26 Val Lys Ile Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15 <210> 27 <211> 16 <212> PRT <213> artificial sequence <220> <223> Human β3 (292-307) <400> 27 Val Lys Val Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Arg Ser Arg 1 5 10 15 <210> 28 <211> 16 <212> PRT <213> artificial sequence <220> <223> Human β4 (287-302) <400> 28 Val Lys Val Ser Ser Pro Lys Val Leu Gln Arg Leu Ile Lys Ser Arg 1 5 10 15

Claims (14)

Hypertension comprising a fusion peptide consisting of a Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide or the amino acid sequence of SEQ ID NO: 19 consisting of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 8 A pharmaceutical composition for prophylaxis or treatment. The pharmaceutical composition for preventing or treating hypertension according to claim 1, wherein the fusion peptide is a Cacb1-derived peptide consisting of the amino acid sequence of SEQ ID NO: 9 and a cell-permeable peptide consisting of the amino acid sequence of SEQ ID NO: 10 fused. The pharmaceutical composition for preventing or treating hypertension according to claim 1, comprising a fusion peptide in which a cell-penetrating peptide consisting of the amino acid sequence of SEQ ID NO: 10 is fused to the Cacb1-derived peptide. The pharmaceutical composition for preventing or treating hypertension according to claim 3, wherein the fusion peptide consists of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 11 to 18. The pharmaceutical composition for preventing or treating hypertension according to claim 1, comprising a fusion peptide in which a half-life extension peptide consisting of the amino acid sequence of SEQ ID NO: 24 is fused to a Cacb1-derived peptide consisting of the amino acid sequence of SEQ ID NO: 4. The pharmaceutical composition for preventing or treating hypertension according to claim 5, wherein the fusion peptide consists of the amino acid sequence of SEQ ID NO: 22. According to claim 1, wherein palmitic acid (Palmitic acid) to the 2nd or 7th lysine (K) of the Cacb1-derived peptide consisting of the amino acid sequence of SEQ ID NO: 4 contains a fatty acid-peptide complex, preventing hypertension or A therapeutic pharmaceutical composition. The pharmaceutical composition for preventing or treating hypertension according to claim 7, wherein the fatty acid-peptide complex comprises any one amino acid sequence selected from the group consisting of SEQ ID NO: 20 or 21. The pharmaceutical for preventing or treating hypertension according to claim 6, comprising a fatty acid-peptide complex in which palmitic acid is bound to the 20th lysine (K) of the fusion peptide consisting of the amino acid sequence of SEQ ID NO: 22. composition. The pharmaceutical composition for preventing or treating hypertension according to claim 9, wherein the fatty acid-peptide complex comprises the amino acid sequence represented by SEQ ID NO: 23. A blood vessel comprising a Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide or a fusion peptide consisting of the amino acid sequence of SEQ ID NO: 19 consisting of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 8 A pharmaceutical composition for relaxation. Hypertension comprising a fusion peptide consisting of a Cacb1 (Voltage-dependent L-type calcium channel subunit beta-1)-derived peptide or the amino acid sequence of SEQ ID NO: 19 consisting of any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 8 A food composition for prevention or improvement.
delete delete

Images