KR20190009139A - Composition for preventing or treating erectile dysfunction comprising proNGF inhibitor - Google Patents

Abstract

The present invention relates to a composition for preventing or treating erectile dysfunction comprising a proNGF (nerve growth factor) inhibitor. According to the present invention, the proNGF inhibitor has an excellent erectile dysfunction improvement effect to increase an activation level of a vascular endothelial cell, a perivascular cell, and nerve cell specific protein to induce regeneration of corpus cavernosum penis endothelial cell and increase penis erectility. Therefore, the proNGF inhibitor can be helpfully used for prevention or treatment of erectile dysfunction.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating erectile dysfunction, comprising proNGF inhibitor,

The present invention relates to a composition for preventing or treating erectile dysfunction comprising a proNGF (nerve growth factor) inhibitor.

Erectile dysfunction is a type of male sexual dysfunction that can not be done because male penis is not erect or erect state is not maintained. The causes of erectile dysfunction are largely classified into psychogenic causes and substrate causes. Cardiogenic erectile dysfunction is caused by excessive secretion of noradrenaline due to excessive action of sympathetic nerves due to psychological and psychological influences, increased smooth muscle tension of the penis cavernosum, and suppression of neurotransmitter secretion.

Substantial erectile dysfunction is classified as neurogenic, vascular, and endocrine erectile dysfunction, depending on its cause. The vascular erectile dysfunction is caused by damage to the corpus cavernosum, vascular endothelial cells, pericytes, and nerve cells due to hyperlipidemia, diabetes, hypertension, smoking, systemic cardiovascular disease, NO), which is caused by a lack of secretion of a relaxant substance.

Recently, studies on erectile dysfunction have focused more on the underlying causes, and oral PDE-5 (phosphodiesterase-5) inhibitors, including Viagra (sildenafil), have been used worldwide for its treatment. These oral drugs are known to increase the concentration of cGMP due to the inhibition of PDE-5, which is specifically distributed in the corpus cavernosum, thereby enhancing the blood flow in the corpus cavernosum, thereby inducing erection and thus treating erectile dysfunction. However, PDE-5 inhibitors such as Viagra have been reported to have several side effects such as headache, flushing, dyspepsia, and cardiac arrest, as well as to modulate transient protein expression and related factors at the molecular level It is not a fundamental treatment. Furthermore, in the case of erectile dysfunction caused by diabetes, such a therapeutic effect is not shown well, and even if it is effective, its efficacy can not be sustained for a long time.

Therefore, there is a need for a remedy for erectile dysfunction which fundamentally treats the normalized corpus cavernosum structure in the erectile dysfunction penis, and the efficacy thereof is prolonged for a long time.

On the other hand, NGF (nerve growth factor) is a nerve growth factor and is mainly involved in the regulation of cell growth, maintenance, proliferation and survival through binding with its Trk receptor. In contrast, proNGF, a precursor of NGF, binds to the p75NTR receptor of the cell membrane and induces and regulates apoptosis through lower signaling (Barcelona et al., Neurobiology of Disease 36: 8826, 2016; Siao et al., Journal of Experimental Medicine 209; 2291, 2012). To date, however, there have been no studies on the proNGF-p75NTR signaling pathway, erectile function, and erectile dysfunction.

Accordingly, the inventors of the present invention conducted a study to develop a composition for preventing or treating erectile dysfunction. As a result, the present inventors confirmed that proNGF inhibitor improves erectile dysfunction by inducing regeneration of penile corpus cavernosum endothelial cells to increase penile erection power , Thereby completing the present invention.

It is an object of the present invention to provide a composition for preventing or treating erectile dysfunction which comprises proNGF (nerve growth factor) inhibitor.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating erectile dysfunction which comprises proNGF expression or activity inhibitor.

The present invention also provides a quasi-drug composition for the prevention or treatment of erectile dysfunction which comprises proNGF expression or activity inhibitor.

The present invention also provides a food composition for preventing or ameliorating erectile dysfunction comprising an inhibitor of proNGF expression or activity.

The proNGF inhibitor according to the present invention has an excellent erectile dysfunction improvement effect by increasing the activity level of vascular endothelial cells, pericytes and nerve cell specific proteins, inducing regeneration of the corpus cavernosum endothelial cells and increasing the erectile power of the penis , And to prevent or treat erectile dysfunction.

FIG. 1 is a graph showing the results of intracoronary cavernosal pressure (erectile force) measurement according to penile nerve electrical stimulation by administration of an anti-proNGF antibody in an erectile dysfunction mouse model.
FIG. 2 shows platelet-endothelial cell adhesion molecule-1 (PECAM-1), which is a vascular endothelial cell-specific protein by administration of anti-proNGF antibody in penile corpus cavernosum tissue of an erectile dysfunction mouse model, Fig. 2 is a graph showing the results of analysis of the activity of NG2 (neuron glial antigen 2), a peripheral cell-specific protein, by a confocal microscope.
FIG. 3 is a graph showing the results of analysis of the activity of beta-III tubulin, a neuron-specific protein by anti-proNGF antibody administration, in a penile corpus callosum tissue of an erectile dysfunction mouse model using a confocal microscope.
FIG. 4 is a graph showing the results of confocal microscopic analysis of the degree of inhibition of vascular endothelial cell death by administration of anti-proNGF antibody in corpus cavernosal tissues of an erectile dysfunction mouse model.

The present invention provides a composition for preventing or treating erectile dysfunction comprising an inhibitor of proNGF expression or activity.

The composition comprises a pharmaceutical composition, a quasi-drug composition or a food composition.

The term " proNGF " in the present invention is known to bind to the p75NTR receptor of the cell membrane as a precursor of the nerve growth factor " nerve growth factor " (NGF) and induce and regulate apoptosis through lower signal transduction. More specifically, " proNGF " may consist of the amino acid sequence shown in SEQ ID NO: 1 or 2 and includes, but is not limited to, functional equivalents of the protein.

The term " proNGF expression or activity inhibitor " in the present invention refers to a substance that has the same meaning as " proNGF inhibitor ", and reduces the mRNA expression or protein activity of proNGF. More specifically, all substances that reduce the expression or activity of proNGF by reducing the expression of proNGF at the transcription level or by interfering with its activity, such as by acting directly on proNGF or acting indirectly on its ligand . The proNGF expression-inhibiting substance can be used in any form such as a compound, a nucleic acid, a peptide, a virus, or a vector containing the nucleic acid, which can inhibit proNGF expression or activity by targeting proNGF. Examples of the substance that inhibits proNGF expression include siRNA, shRNA, miRNA, antisense oligonucleotide, ribozyme, DNAzyme, PNA (peptide nucleic acids), etc. which specifically bind to proNGF mRNA and inhibit the expression of proNGF mRNA , But is not limited thereto. Examples of the substance that inhibits proNGF activity include an antibody or an antigen-binding fragment thereof, an aptamer, a compound, etc. that specifically binds to the protein of proNGF and inhibits the activity of proNGF protein, It does not.

As used herein, the term " small interference RNA " (siRNA) is a nucleic acid molecule capable of mediating RNA interference or gene silencing and refers to a small RNA fragment of 21-25 nucleotide size. The siRNA of the present invention may have a structure in which a sense strand (a corresponding sequence corresponding to an mRNA sequence) and an antisense strand (a sequence complementary to an mRNA sequence) are located on opposite sides to form a double strand, single-stranded structure with sense and antisense strands. The siRNA of the present invention is not limited to the complete pairing of the double-stranded RNA portions constituting the pair of RNAs, but may be a mismatch (the corresponding base is not complementary), a bulge (no base corresponding to one of the chains) Non-paired portions may be included.

The term " shRNA (short hairpin RNA) " in the present invention is intended to overcome the disadvantages of high cost biosynthesis cost of siRNA, short term maintenance of RNA interference effect due to low cell transfection efficiency, Viruses, lentiviruses, and plasmid expression vector systems to introduce them into cells and express them. It is well known that such shRNAs are converted into siRNAs with correct structure by the siRNA processing enzyme (Dicer or Rnase Ⅲ) present in the cell to induce silencing of the target gene.

As used herein, the term " miRNA (microRNA) " is a single strand RNA molecule of 21-25 nucleotides and is a regulatory substance that controls gene expression of eukaryotic organisms through inhibition in the step of disruption or detoxification of target mRNA. These miRNAs are made up of two stages of processing. The first miRNA transcript is made in the nucleus by the RNase III type enzyme called Drosha, which is made into a stem-loop structure of about 70-90 bases, that is, a pre-miRNA, which is then transferred to the cytoplasm and transferred to an enzyme called Dicer To produce a mature miRNA of 21-25 bases. These miRNAs complementarily bind to the target mRNA and act as post-transcriptional gene suppressors, leading to translation inhibition and mRNA destabilization. miRNAs are involved in a variety of physiological phenomena and diseases.

The term " antisense oligonucleotide " in the present invention means a DNA or RNA or a derivative thereof containing a nucleic acid sequence complementary to the sequence of a specific mRNA, which binds to a complementary sequence in the mRNA, .

As used herein, the term " antibody " refers to a specific protein molecule, as is known in the art, directed against an antigenic site. For purposes of the present invention, an antibody refers to an antibody that specifically binds to proNGF, which is obtained by cloning each gene into an expression vector according to a conventional method to obtain a protein encoded by the gene, ≪ / RTI > Also included are partial peptides that can be made from the protein, and the partial peptides of the invention include at least 7 amino acids, preferably 9 amino acids, more preferably 12 or more amino acids. The form of the antibody of the present invention is not particularly limited, and any polyclonal antibody, monoclonal antibody or antigen-binding antibody thereof may be included in the antibody of the present invention and include all immunoglobulin antibodies. Furthermore, the antibodies of the present invention include special antibodies such as humanized antibodies. Antibodies used in the present invention include functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having at least an antigen-binding function, and includes Fab, F (ab ') 2, F (ab') 2 and Fv.

In the present invention, the term " aptamer " means a nucleic acid molecule having a binding activity to a predetermined target molecule. The aptamer may be RNA, DNA, modified nucleic acid or a mixture thereof, and may be in the form of a linear or cyclic form. The oligonucleotide library called SELEX (systematic evolution of ligands by exponential enrichment) It is a substance obtained by separating oligomers that bind with high affinity and selectivity to specific chemical molecules or biological molecules by an evolutionary method. The aptamer can specifically bind to the target and modulate the activity of the target, for example, by blocking the ability of the target to function through binding.

The term " erectile dysfunction " in the present invention means a state in which erectile function is not sufficiently maintained or maintained in the sex life, and erectile dysfunction is generally defined as a state in which such state lasts for 3 months or more. The erectile dysfunction can be broadly divided into psychogenic erectile dysfunction caused by psychological factors and organic erectile dysfunction caused by physical disability. The proNGF inhibitor of the present invention ultimately improves the function of the penis, Can be treated. The erectile dysfunction may be due to a disease such as diabetes, penile tissue damage, coronary artery disease, nephropathy, neuropathy, hypertension, arteriosclerosis or hyperlipemia, but is not limited to the above example, Any disease is included without limitation.

In the present invention, the term " corpuscle " is an erectile tissue constituting the main body of the mammalian penis or clitoris. The corpuscle is piled up with a membrane of a thick and durable connective tissue containing elastic fibers. . Within the male penis, there are two penile cavernosomes on the left and right, and one urethral cavernosum on the lower side. There are clitoral cavernosomes and urethral cavernosomes with similar structures to the penis cavernosum in women. When the cavernosum is full of venous blood, the penis becomes erect. The penile cavernosal smooth muscle and penile blood vessels are usually contracted by adrenergic sympathetic nerves, and after erectile response due to relaxation, they return to contracted state by stimulation of the adrenergic sympathetic nervous system. Although erectile tissue maintains normal contraction state, erectile tissue is relaxed by stimulation of the nervous system, various neurotransmitters and hormones in the body and erectile tissues, erection is started, erection is started, maintain. After this erection reaction is completed, the erectile tissue is maintained in a contracted state.

The term " prophylactic " in the present invention refers to any action that inhibits erectile dysfunction or inhibits or delays erectile dysfunction using a composition comprising a proNGF inhibitor of the present invention.

The term " treatment " in the present invention refers to any action that improves or benefits erectile dysfunction using a composition comprising a proNGF inhibitor of the present invention.

The proNGF inhibitor according to the present invention has an effect of improving erectile dysfunction by increasing the activity level of vascular endothelial cells, pericytes and nerve cell specific proteins, inducing regeneration of the corpus cavernosum endothelial cells and increasing the erectile function of the penis.

As described above, the proNGF inhibitor according to the present invention is excellent in prevention, improvement and therapeutic effect of erectile dysfunction, and thus can be used as medicines, quasi-drugs or health functional foods useful for prevention or treatment of erectile dysfunction.

When the composition of the present invention is formulated into a pharmaceutical composition for the purpose of preventing or treating erectile dysfunction, it may further comprise suitable carriers, excipients and diluents conventionally used in the preparation of pharmaceutical compositions. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA.

In addition, the pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method . When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term " administering " as used herein means providing the subject invention with a composition of the invention in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. The composition may be administered once a day or divided into several doses.

The pharmaceutical composition of the present invention may be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intradiscal injection. However, at the time of oral administration, since the protein is digested, it is preferable to formulate the oral composition so as to coat the active agent or protect it from decomposition at the top. The composition of the present invention can preferably be administered in the form of an injection.

The composition of the present invention may contain one or more known active ingredients having proinfection or prophylactic effects of erectile dysfunction together with a proNGF inhibitor.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for prevention and treatment of erectile dysfunction.

In addition, the composition of the present invention may be added to a quasi-drug composition for the purpose of preventing or treating erectile dysfunction.

When the composition of the present invention is used as a quasi-drug composition, the active ingredient may be added as it is or may be used together with other quasi-drug components, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).

In addition, the composition of the present invention may be added to a health functional food for the purpose of preventing or improving erectile dysfunction.

The term " health functional food " in the present invention refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease and aging inhibition.

The health functional food may include food-acceptable food supplementary additives, and may further include suitable carriers, excipients and diluents conventionally used in the production of health functional foods.

When the composition of the present invention is used as a food additive, the above-mentioned active ingredient can be directly added or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the composition of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, it 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 exceeding the above range.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The redundant contents are taken into consideration in the complexity of the present specification, and terms not otherwise defined herein have the meanings commonly used in the art to which the present invention belongs.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example  1. Design of erectile dysfunctional animal models proNGF  Administration of inhibitors

The mice were divided into three groups (N = 5 / group; group 1, normal mice; group 2), streptomycin Mice injected with PBS (phosphate-buffered solution) [20 [mu] L) to the corpus cavernosum + group 3, streptozotocin Diabetic induction with gonadotropin (8 weeks after intraperitoneal injection at 50 mg / kg for 5 consecutive days) + penile corpus callosum twice daily with anti-proNGF antibody [day -3,0; 20 μL]).

As an inhibitor for inhibiting the activity of proNGF, an anti-proNGF antibody was used (Biosensis, S-080-100) in which a prodomain fragment of human NGF was used as an antigen and a cross reaction against mouse and rat proNGF was confirmed.

Example  2. In an erectile dysfunctional animal model, anti- proNGF  Efficacy of erectile dysfunction treatment of antibodies

2-1. Measurement of erectile force by electric stimulation

In order to confirm the effect of the proNGF inhibitor according to the present invention on improving the erectile potency, the erectile force was confirmed by stimulating the nerve of the corpus cavernosum. More specifically, as in Example 1, after 2 weeks of administration of the anti-proNGF antibody to the corpus cavernosum, the left epidermis was covered in the lower abdomen of each mouse, and the penile cavernosal nerve located at the posterior lateral side of the prostate ) Were prepared to look good. After the catheter was connected to the penis nerve with a pressure transducer (BioSpec System, USA) connected, a platinum electrode was placed on the penis nerve to give electrical stimulation to the penis nerve. Was added with electrical stimulation. Thus, the intracavital pressure (ICP) measured through the catheter inserted into the penis was measured. The curves for the corpus cavernosal pressure are shown in Fig. 1A, the maximal ICP (maximal ICP) is shown in Fig. 1B, and the area (total ICP [area under the curve] As shown in Fig. 1C. The ICP is an index indicating the erectile power, and the electric stimulation period for 1 minute is indicated by black bars on the horizontal axis in FIG.

As shown in FIG. 1, the anti-proNGF antibody-treated group (pro-NGF), which is a proNGF inhibitor, showed an erectile potency recovery to a control level and remarkably improved erectile function compared to the PBS-treated group. In particular, we found that two parameters related to erectile function of the penis, namely the maxillary intima pressure and the area of the penile pressure curve, were significantly increased by the administration of anti-proNGF antibody.

2-2. In vascular endothelial tissue, Pericyte  And neuron-specific protein expression

The penis consists of cavernosal tissues that function as a penis erection and small blood vessels that supply nutrition. However, under pathological conditions such as diabetes, hypertension, nerve damage, etc., lesions are formed in the corpus cavernosum including these blood vessels and the structure of the corpuscle becomes abnormal, resulting in erectile dysfunction. In order to confirm the effect of proNGF inhibitor according to the present invention on the erectile dysfunction treatment under the above background, it is known that the platelet-endothelial cell adhesion molecule-1 (PECAM-1), which is a specific protein for vascular endothelial cells, NG2, a specific protein in cells, and beta III tubulin, a specific protein in neurons.

More specifically, the penile corpus cavernosum was separated from each mouse after 2 weeks of administration of the anti-proNGF antibody to the corpus cavernosum as in Example 1, and fixed in 4% paraformaldehyde for 24 hours (at 4 ° C ). The corpus cavernosum tissues were fixed using a freezing foraging agent, and then cut into a thickness of 7 mu m in a frozen section to prepare a penile tissue section. Next, prepared penile tissue sections were placed on slides and fixed in 4% paraformaldehyde for about 5 minutes. Fixed penis tissue sections were washed three times with wash buffer (2% FBS + 0.1% Sodium Azide in PBS) and blocked with non-specific protein blocking buffer (5% BSA in PBS) for 1 hour. Thereafter, the cells were reacted at 4 ° C for 16 hours with a primary antibody (anti-PECAM-1 hamster antibody 1:50, anti-NG2 rat antibody 1:50, anti-βIII tubulin antibody 1: 200) (TRITC-labeled anti-hamster antibody or FITC-labeled anti-rat antibody or TRITC-labeled anti-chicken antibody 1: 1000), followed by washing three times with washing buffer to remove remaining antibodies. Were reacted at room temperature for 2 hours. After the reaction was completed, the cells were washed twice with a washing buffer to remove remaining antibodies, and then analyzed using a confocal microscope. The results are shown in Fig. 2 and Fig.

2 and 3, platelet-endothelial cell adhesion molecule-1 (PECAM-1: platelet-specific), which is a vascular endothelial cell-specific protein as compared to a normal control group, / Endothelial Cell Adhesion Molucule-1), neuron glial antigen 2 (NG2) specific for pericytes, and beta III tubulin, a neuron-specific protein, but the proNGF inhibitor anti-proNGF antibody (DM + anti-proNGF) increased the activity of PECAM-1, NG2, and beta-III tubulin. This suggests that the proNGF inhibitor of the present invention is not merely a temporary improvement in erectile dysfunction due to an increase in blood flow, but is a result of stimulation of the corpus cavernosum tissue This is the result of proving that the erectile dysfunction treatment effect is obtained by solving the root cause of the improvement of the condition itself.

2-3. Inhibition of vascular endothelial cell apoptosis in corpus cavernosal tissue

In order to confirm the effect of erectile dysfunction treatment by the proNGF inhibitor according to the present invention, the effect on the apoptosis of vascular endothelial cells was confirmed. More specifically, the penile corpus cavernosum was separated from each mouse after 2 weeks of administration of the anti-proNGF antibody to the corpus cavernosum as in Example 1, and fixed in 4% paraformaldehyde at 4 ° C for 24 hours . The above tissues were fixed using a frozen foraging agent, and then cut into a thickness of 7 mu m in a frozen section to prepare a penile tissue section. Next, prepared penile tissue sections were placed on slides and fixed in 4% paraformaldehyde for about 5 minutes. Fixed penile tissue sections were washed three times with wash buffer (PBS) and TUNEL staining was performed following the experimental instructions of the TUNEL Kit (S7111 | ApopTag® Plus In Situ Apoptosis Fluorescein Detection Kit). TUNEL staining followed by three washes with wash buffer (PBS) followed by blocking with non-specific protein blocking buffer (5% BSA in PBS) for 1 hour. Thereafter, the cells were reacted with anti-PECAM-1 hamster antibody 1:50 for 16 hours at 4 DEG C, washed three times with washing buffer to remove the remaining antibodies, and then washed with secondary TRITC-labeled anti-hamster Antibody 1: 1000) at room temperature for 2 hours. After the reaction was completed, the cells were washed twice with a washing buffer to remove remaining antibodies, and then analyzed using a confocal microscope. The results are shown in Fig.

As shown in FIG. 4, the number of killed vascular endothelial cells was significantly increased in the penile corpus cavernosum tissues of the PBS-administered group (DM + PBS) compared to the control group (control), but the proNGF inhibitor anti-proNGF antibody DM + anti-proNGF) was significantly reduced in the corpus cavernosum tissues. This is a result of demonstrating that proinflammatory inhibitor inhibits the apoptosis of vascular endothelial cells, which is increased by diabetes, and thereby restores erectile power.

These results suggest that proNGF inhibitor according to the present invention increases the activity level of vascular endothelial cells, pericytes and nerve cell specific proteins to induce regeneration of endothelial cells of the corpus cavernosum and increase erectile dysfunction of the penis, And it was confirmed that it had improvement effect. Therefore, the proNGF inhibitor according to the present invention can be usefully used for prevention or treatment of erectile dysfunction.

Hereinafter, examples of pharmaceutical compositions and food compositions according to the present invention will be described, but the present invention is not to be construed as limiting the present invention .

Formulation example  1. Preparation of pharmaceutical compositions

1-1. Preparation of capsules

proNGF inhibitor 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

1-2. Injection preparation

proNGF inhibitor 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

1-3. Liquid  Produce

proNGF inhibitor 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example  2. Preparation of food composition

2-1. Manufacture of health food

proNGF inhibitor 100 mg

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

<110> Inha University Research and Business Foundation <120> Composition for preventing or treating erectile dysfunction          comprising proNGF inhibitor <130> 1-246 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 241 <212> PRT <213> Homo sapiens <400> 1 Met Ser Met Leu Phe Tyr Thr Leu Ile Thr Ala Phe Leu Ile Gly Ile   1 5 10 15 Gln Ala Glu Pro His Ser Glu Ser Asn Val Pro Ala Gly His Thr Ile              20 25 30 Pro Gln Ala His Trp Thr Lys Leu Gln His Ser Leu Asp Thr Ala Leu          35 40 45 Arg Arg Ala Arg Ser Ala Pro Ala Ala Ala Ile Ala Ala Arg Val Ala      50 55 60 Gly Gln Thr Arg Asn Ile Thr Val Asp Pro Arg Leu Phe Lys Lys Arg  65 70 75 80 Arg Leu Arg Ser Pro Arg Val Leu Phe Ser Thr Gln Pro Pro Arg Glu                  85 90 95 Ala Ala Asp Thr Gln Asp Leu Asp Phe Glu Val Gly Gly Ala Ala Pro             100 105 110 Phe Asn Arg Thr His Arg Ser Ser Arg Ser Ser Ser His Pro Ile Phe         115 120 125 His Arg Gly Glu Phe Ser Val Cys Asp Ser Val Ser Val Trp Val Gly     130 135 140 Asp Lys Thr Thr Ala Thr Asp Ile Lys Gly Lys Glu Val Met Val Leu 145 150 155 160 Gly Glu Val Asn Ile Asn Asn Ser Val Phe Lys Gln Tyr Phe Phe Glu                 165 170 175 Thr Lys Cys Arg Asp Pro Asn Pro Val Asp Ser Gly Cys Arg Gly Ile             180 185 190 Asp Ser Lys His Trp Asn Ser Tyr Cys Thr Thr Thr His Thr Phe Val         195 200 205 Lys Ala Leu Thr Met Asp Gly Lys Gln Ala Ala Trp Arg Phe Ile Arg     210 215 220 Ile Asp Thr Ala Cys Val Cys Val Leu Ser Arg Lys Ala Val Arg Arg 225 230 235 240 Ala     <210> 2 <211> 241 <212> PRT <213> Mus musculus <400> 2 Met Ser Met Leu Phe Tyr Thr Leu Ile Thr Ala Phe Leu Ile Gly Val   1 5 10 15 Gln Ala Glu Pro Tyr Thr Asp Ser Asn Val Pro Glu Gly Asp Ser Val              20 25 30 Pro Glu Ala His Trp Thr Lys Leu Gln His Ser Leu Asp Thr Ala Leu          35 40 45 Arg Arg Ala Arg Ser Ala Pro Thr Ala Pro Ile Ala Ala Arg Val Thr      50 55 60 Gly Gln Thr Arg Asn Ile Thr Val Asp Pro Arg Leu Phe Lys Lys Arg  65 70 75 80 Arg Leu His Ser Pro Arg Val Leu Phe Ser Thr Gln Pro Pro Thr                  85 90 95 Ser Ser Asp Thr Leu Asp Leu Asp Phe Gln Ala His Gly Thr Ile Pro             100 105 110 Phe Asn Arg Thr His Arg Ser Ser Arg Ser Ser Thr His Pro Val Phe         115 120 125 His Met Gly Glu Phe Ser Val Cys Asp Ser Val Ser Val Trp Val Gly     130 135 140 Asp Lys Thr Thr Ala Thr Asp Ile Lys Gly Lys Glu Val Thr Val Leu 145 150 155 160 Ala Glu Val Asn Ile Asn Asn Ser Val Phe Arg Gln Tyr Phe Phe Glu                 165 170 175 Thr Lys Cys Arg Ala Ser Asn Pro Val Glu Ser Gly Cys Arg Gly Ile             180 185 190 Asp Ser Lys His Trp Asn Ser Tyr Cys Thr Thr Thr His Thr Phe Val         195 200 205 Lys Ala Leu Thr Thr Asp Glu Lys Gln Ala Ala Trp Arg Phe Ile Arg     210 215 220 Ile Asp Thr Ala Cys Val Cys Val Leu Ser Arg Lys Ala Thr Arg Arg 225 230 235 240 Gly    

Claims (6)

A prophylactic or therapeutic agent for erectile dysfunction comprising an expression or activity inhibitor of proNGF (nerve growth factor). The pharmaceutical composition for preventing or treating erectile dysfunction according to claim 1, wherein the expression inhibitor is at least one selected from the group consisting of miRNA, siRNA, shRNA and antisense oligonucleotide that specifically binds to mRNA of proNGF . The pharmaceutical composition for preventing or treating erectile dysfunction according to claim 1, wherein the activity inhibitor is an antibody that specifically binds to the protein of proNGF. The pharmaceutical composition for preventing or treating erectile dysfunction according to claim 1, wherein the proNGF expression or activity inhibitor inhibits the apoptosis of the corpus cavernosal vascular endothelial cells. A prophylactic composition for preventing or treating erectile dysfunction, which comprises an expression or activity inhibitor of proNGF (nerve growth factor). proNGF (nerve growth factor) expression or activity inhibitor.

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