KR102592438B1 - Reagent composition for inhibiting AR and 5AR2 of androgen signaling related factors - Google Patents

Abstract

The present invention relates to a composition for preventing, improving, or treating benign prostatic hyperplasia or androgenic alopecia, comprising Centella asiatica extract as an active ingredient. As a result, it is a natural material that is harmless to the human body and can minimize side effects, and is especially effective in suppressing the expression of factors related to androgen signaling.

Description

Reagent composition for inhibiting AR and 5AR2 expression of androgen signaling related factors {Reagent composition for inhibiting AR and 5AR2 of androgen signaling related factors}

The present invention relates to a composition for preventing, improving or treating benign prostatic hyperplasia or androgenic alopecia.

Prostatic hyperplasia is a common chronic disease of the urinary tract in men over 50 years of age (Berry S.J. et al., 1984). Prostatic hyperplasia is also associated with lower urinary tract symptoms (LUTS), including increased urinary intermittency, increased urinary frequency, urinary emergencies, impaired urinary flow, and incomplete bladder emptying, which impairs quality of life in older adults and negatively impacts daily life. (Sarma, A.V. et al., 2012).

Although the underlying cause of benign prostatic hyperplasia is unknown, it is well known that the prostate in aging men is affected by androgens (Kim S.K. et al., 2015). During the progression of prostatic hypertrophy, androgens promote the proliferation of epithelial or stromal cells in the prostate in an autocrine or paracrine manner, resulting in an imbalance in prostate cell proliferation and apoptosis. This has been considered an important cause of BPH (Choi, H.M. et al., 2016).

In particular, dihydrotestosterone (DHT) is well known to be associated with the development of benign prostatic hyperplasia. As we age, the male sex hormone testosterone begins to be converted to dihydrotestosterone (DHT) at a high rate in the prostate. This is primarily due to the concentration of reductase, an enzyme that converts testosterone to DHT. The reason is that is rising. DHT is a more potent androgen compared to TST due to its high binding affinity to the androgen receptor (AR). 5α reductase type-2 (5AR2) converts TST to DHT in the prostate, and DHT binds to AR to increase transcription of androgen-dependent genes, ultimately leading to stimulation of protein synthesis associated with prostate hyperplasia. -Improves specific antigen (PSA) levels. Meanwhile, PSA levels increase during the progression of BPH and prostate cancer, which is why PSA is widely used in the diagnosis of BPH (Chen, Y. et al., 1996).

The cell cycle occurs in prostate stromal and epithelial cells, leading to their division and replication. G1/S progression is highly regulated by cyclin D1. Additionally, proliferating cell nuclear antigen (PCNA) is an acidic nuclear protein recognized as a histological marker for the G1/S phase of the cell cycle. Therefore, the expression of PCNA and cyclin D1 may reflect the proliferative state of prostate cells during BPH (Wang, W., et al., 2004).

Conversely, apoptosis of the prostatic epithelium occurs more frequently in the prostate under normal conditions than in benign prostatic hyperplasia. Bcl-2, which inhibits apoptosis, is also found in the prostatic epithelium, leading to proliferation of prostatic tissue (Cardillo, M., et al., 1997).

Another symptom caused by male hormones such as dihydrotestosterone (DHT) is male pattern baldness. The causes of hair loss have been suggested to include poor blood circulation, excessive male hormone action, excessive sebum secretion, decreased scalp function due to dandruff and other bacteria, genetic factors, and aging stress. However, until now, excessive male hormone secretion has been suggested as the cause. The cause of androgenic alopecia, the most common type of hair loss directly related to hair loss, is being revealed.

In cases of androgenetic alopecia, healthy hair gradually becomes thinner, shorter, and becomes weaker and more easily broken. This phenomenon is called miniaturization. Hair follicles that undergo miniaturization eventually become thin, hard to see, and turn into short downy hair, leading to hair loss. Accordingly, many studies have recently been reported on the prevention and treatment of hair loss by suppressing male hormone activity.

Testosterone is converted into dihydrotestosterone (DHT), an active male hormone, by 5α-reductase, and this activated dihydrotestosterone binds to androgen receptors to promote protein synthesis in hair follicle cells. Delaying this shortens the growth period of hair follicles and causes hair follicles to atrophy, causing hair loss. Additionally, sebum may be excessively produced during androgenetic alopecia, and hair loss accompanied by inflammation may occur on the scalp as a result (Dennis A. Holt, et. al,. 1990).

Currently developed treatments for benign prostatic hyperplasia, such as alpha-1 adrenergic blockers such as terazosin, doxazosin, tamsulasin, and alfuzosin, relieve symptoms of urethral obstruction by relaxing prostatic smooth muscle, but block sympathetic nerves, causing hypotension and vasodilation. Side effects such as headaches have been reported.

Finasteride (Fi) and Dutasteride, which are 5α reductase inhibitors, are known to be effective in reducing prostate size by inhibiting the conversion of testosterone to the active metabolite dihydrotestosterone (DHT) by 5α reductase and are used for BPH and alopecia. However (Park, E.S., et al., 2018), these drugs are known to cause side effects such as erectile dysfunction, decreased libido, and decreased semen volume in the ejaculate. Although currently used drug treatments are somewhat effective, their side effects are problematic, and in fact, benign prostatic hyperplasia is one of the persistent aging phenomena that makes fundamental treatment difficult.

Therefore, researchers looking for effective treatment strategies with fewer side effects are increasingly interested in alternative medicine, including drugs made from natural ingredients. Currently, among natural ingredients, only saw palmetto extract is widely known as a functional food with an effect on benign prostatic hyperplasia (Marks, L.S., et al., 2000).

Korean Patent No. 10-1881417 Korean Patent No. 10-2017187

The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating benign prostatic hyperplasia or androgenic alopecia, which contains as an active ingredient a centella asiatica extract, which is a natural material that is harmless to the human body and can minimize side effects.

The purpose of the present invention is to provide a food composition for preventing or improving benign prostatic hyperplasia or androgenic alopecia, which contains as an active ingredient Centella asiatica extract, which is a natural material that is harmless to the human body and can minimize side effects.

According to one aspect of the invention,

A pharmaceutical composition for preventing or treating benign prostatic hyperplasia or androgenetic alopecia is provided, comprising Centella asiatica extract as an active ingredient.

The Centella asiatica extract may be extracted with water, alcohol with 1 to 4 carbon atoms, or a mixed solvent thereof.

The pharmaceutical composition may be used to inhibit the expression of factors related to androgen signaling.

The androgen signaling-related factor may be any one selected from SRC-1 (steroid receptor coactivator), AR (androgen receptor), 5AR2 (5alpha reductase), and PSA (prostate specific antigen).

The pharmaceutical composition may be used to inhibit the expression of growth factors in the prostate.

The growth factor within the prostate may be any one selected from Epidermal growth factor (EGF), Insulin like growth factor I (IGF-1), Transforming growth factor beta 1 (TGF-1β), and Vascular endothelial growth factor (VEGF). .

The pharmaceutical composition may be used to inhibit the expression of PI3K/Akt and HIF-1 signaling related factors.

The PI3K/Akt and HIF-1 signaling-related factors may be any one selected from PI3K (phosphatidylinositol-3-kinase), Akt (protein kinase B), and HIF-1 (Hypoxia-inducible factors).

The pharmaceutical composition may be used to inhibit the expression of factors related to cell proliferation in prostate tissue.

The cell proliferation-related factor in the prostate tissue may be PCNA (Proliferating Cell Nuclear Antigen) or Cyclin D1.

According to another aspect of the present invention,

A food composition for preventing or improving benign prostatic hyperplasia or androgenic alopecia is provided, containing Centella asiatica extract as an active ingredient.

According to another aspect of the present invention,

Provided is a health functional food for preventing or improving benign prostatic hyperplasia or androgenic alopecia, comprising a food composition containing Centella asiatica extract as an active ingredient.

The pharmaceutical composition for preventing or treating benign prostatic hyperplasia or androgenetic alopecia containing the centella asiatica extract of the present invention as an active ingredient is a natural material that is harmless to the human body and can minimize side effects, and especially inhibits the expression of factors related to androgen signaling. There is an effect that can be done.

The food composition for preventing or improving benign prostatic hyperplasia or androgenetic alopecia containing the centella asiatica extract of the present invention as an active ingredient is a natural ingredient that is harmless to the human body and can minimize side effects, and especially inhibits the expression of factors related to androgen signaling. There is an effect that can be done.

Figure 1 shows the results of Western blot according to Experimental Example 1.
Figure 2 shows the results of measuring AR protein expression level according to Experimental Example 1.
Figure 3 shows the results of measuring 5AR2 protein expression level according to Experimental Example 1.
Figure 4 shows the results of measuring PSA protein expression level according to Experimental Example 1.
Figure 5 shows the results of comparing photos of the prostates of each experimental group separated according to Experimental Example 2 and the weight of the prostates.
Figure 6 shows the prostate index measurement results according to Experimental Example 2.
Figure 7 shows microscopic pictures of prostate epithelial cells and the results of measuring epithelial cell thickness according to Experimental Example 2.
Figure 8 shows the results of analysis of the ability to suppress the expression of androgen signaling-related factors according to Experimental Example 3.
Figure 9 is a representative diagram of the references in Experimental Example 4.
Figure 10 shows the results of analysis of growth factor expression inhibition ability according to Experimental Example 4.
Figure 11 is a schematic diagram of the PI3K/Akt and HIF 1 signaling pathways.
Figure 12 shows the results of a Western blot experiment according to Experimental Example 5.
Figure 13 shows the results of analysis of the ability to suppress expression of factors related to cell proliferation in prostate tissue according to Experimental Example 6.

Hereinafter, the present invention will be described in detail.

The pharmaceutical composition for preventing or treating benign prostatic hyperplasia or androgenetic alopecia of the present invention includes Centella asiatica extract as an active ingredient.

Centella asiatica is a plant in the Apiaceae family that blooms white or light purple flowers on small fan-shaped leaves with serrated edges. In India, after seeing a wounded tiger rolling around in a place with a lot of Centella asiatica to heal, it was called tiger plant and has been used as medicine for a long time.

The extraction solvent for extracting the Centella asiatica extract is water, lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof. The lower alcohol may include 20 to 80% methanol, ethanol, butanol, or propanol.

The extraction solvent is not particularly limited, but an extract extracted with a 60 to 80% aqueous ethanol solution is preferred.

The term 'extract' used in this specification while referring to the Centella asiatica extract includes not only the crude extract obtained by treatment with an extraction solvent, but also the processed product of the Centella asiatica extract. For example, Centella asiatica extract can be prepared in powder form by additional processes such as reduced pressure distillation and freeze-drying or spray drying.

In addition, the Centella asiatica extract of the present invention, in a broad sense, also includes Centella asiatica processed products formulated so that they can be administered to animals, such as Centella asiatica powder. Although experiments were conducted with centella asiatica in the present invention, those skilled in the art would be able to predict that the desired effect can be achieved even in the form of a centella asiatica processed product.

Meanwhile, in this specification, the term 'including as an active ingredient' means containing a sufficient amount to achieve the efficacy or activity of the Centella asiatica extract or fractions thereof. For example, the Centella asiatica extract or its fraction is used at a concentration of 10 to 1500 μg/ml, preferably 100 to 1000 μg/ml. Since the Centella asiatica extract or its fraction is a natural product and does not cause side effects to the human body even when administered in excessive amounts, the upper quantitative limit of the Centella asiatica extract or its fraction included in the composition of the present invention can be selected within an appropriate range by a person skilled in the art.

In addition, the present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia or androgenic alopecia, comprising Centella asiatica extract as an active ingredient.

The pharmaceutical composition can be used to inhibit the expression of factors related to androgen signaling.

The androgen signaling-related factor may be any one selected from SRC-1 (steroid receptor coactivator), AR (androgen receptor), 5AR2 (5alpha reductase), and PSA (prostate specific antigen).

The pharmaceutical composition can be used to inhibit the expression of growth factors in the prostate.

The growth factor within the prostate may be any one selected from Epidermal growth factor (EGF), Insulin like growth factor I (IGF-1), Transforming growth factor beta 1 (TGF-1β), and Vascular endothelial growth factor (VEGF). .

The pharmaceutical composition can be used to inhibit the expression of PI3K/Akt and HIF-1 signaling related factors.

The PI3K/Akt and HIF-1 signaling-related factors may be any one selected from PI3K (phosphatidylinositol-3-kinase), Akt (protein kinase B), and HIF-1 (Hypoxia-inducible factors).

The pharmaceutical composition can be used to inhibit the expression of factors related to cell proliferation in prostate tissue.

The cell proliferation-related factor in the prostate tissue may be PCNA (Proliferating Cell Nuclear Antigen) or Cyclin D1.

The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable auxiliaries in addition to the active ingredients, and the auxiliaries include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, and lubricants. Agents or flavoring agents can be used.

For administration, the pharmaceutical composition may be preferably formulated as a pharmaceutical composition containing one or more pharmaceutically acceptable carriers in addition to the active ingredients described above.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, etc. Additionally, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacance or sodium oleate, sodium stearate, magnesium stearate, sodium Includes benzoate, sodium acetate, sodium chloride, etc. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, etc.

Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these ingredients can be mixed and used, and other common additives such as antioxidants, buffers, and bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., and is preferably administered orally.

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and is usually A skilled doctor can easily determine and prescribe an effective dosage for desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention can be prepared in unit dose form by formulating using a pharmaceutically acceptable carrier and/or excipient, or can be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, or capsule, and may additionally contain a dispersant or stabilizer.

In addition, the present invention provides a food composition for preventing or improving benign prostatic hyperplasia or androgenic alopecia, comprising Centella asiatica extract as an active ingredient.

The food composition according to the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, and health supplements. etc.

The food composition of the present invention may contain not only Centella asiatica extract or fractions thereof as an active ingredient, but also ingredients commonly added during food production, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Includes. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured as a drink or beverage, in addition to the Centella asiatica extract or fractions thereof of the present invention, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included. You can.

The present invention provides a health functional food containing a food composition for preventing or improving benign prostatic hyperplasia or androgenic alopecia, containing Centella asiatica extract as an active ingredient. Health functional foods are foods manufactured by adding Centella asiatica extract or its fractions to food ingredients such as beverages, teas, spices, gums, and confectionery, or by encapsulating, powdering, or suspending them, and when consumed, they have specific health effects. However, unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of Centella asiatica extract added in such health functional foods cannot be uniformly specified as it depends on the type of health functional food being targeted. However, it can be added within a range that does not damage the original taste of the food, and is usually 0.01% for the target food. to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets, or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule, or beverage.

In addition, the present invention provides the use of Centella asiatica extract for the manufacture of medicine or food for preventing, treating or improving benign prostatic hyperplasia or androgenetic alopecia.

Additionally, the present invention provides a method for improving, preventing, or treating atopy comprising administering an effective amount of Centella asiatica extract to a mammal.

As used herein, the term “mammal” refers to a mammal, preferably a human, that is the subject of treatment, observation or experiment.

As used herein, the term "effective amount" means the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal, or human, as believed by a researcher, veterinarian, physician, or other clinician, to cause the disease in question. or an amount that induces relief of symptoms of the disorder. The effective amount and frequency of administration of the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by a person skilled in the art, and can be determined based on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, and the patient's age, weight, and general health. It can be adjusted according to various factors, including condition, gender and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs. In the prevention, treatment or improvement method of the present invention, for adults, it is preferable to administer the extract at a dose of 0.001 g/kg to 10 g/kg once or several times a day.

In the treatment method of the present invention, the composition containing Centella asiatica extract as an active ingredient is administered through conventional routes through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, or intradermal routes. It can be administered in any way.

Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are merely illustrative of the present invention, and it is clear to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the attached patent claims.

[Example]

Example 1: Preparation of Centella asiatica extract

Dried commercially sold Indonesian centella asiatica ( Centella asiatica ) was purchased, 1 L of DW was added to a 100 g sample, and extraction was performed in a constant temperature water bath at 100°C for 1 hour. This extract was filtered through filter paper and then concentrated under reduced pressure using a vacuum rotary concentrator. Afterwards, it was cooled in a deep freezer for a day and freeze-dried. Freeze-dried Centella asiatica hot water extract was dissolved in DMSO (dimethyl sulfoxide) solvent at a concentration of 1 g/ml and used in cell experiments, and dissolved in DW and used in animal experiments.

[Experimental example: in vitro experiment]

Experimental Example 1: Analysis of ability to suppress expression of androgen signaling-related factors

LNCaP cells were treated with testosterone (testosterone propionate, TP) to induce cell proliferation, and after treatment with the Centella asiatica extract of Example 1, AR (androgen receptor), 5AR2 (5 alpha reductase), and PSA (prostate specific antigen) ) was confirmed to suppress the expression of .

Specifically, the inhibitory ability of androgen signaling-related factors of Centella asiatica was confirmed in LNCaP cells. LNCaP cells were seeded in a ⁶ -well plate at 2 Treated for 24 hours. At this time, the group treated with 1㎍/㎖ of finasteride (Fi), an androgen inhibitor, was used as a positive control group.

After the cells were treated, the expression levels of androgen signaling-related factors AR, 5AR2, and PSA proteins were measured using Western blotting, and the band intensity was determined by setting the area of each sample using image J (NIH ver. 1.48, USA). Then, the relative area of β-Actin was set in the same way and quantified by calculating by dividing the value. The results of the Western blot experiment according to this are shown in Figure 1, the results of measuring AR protein expression levels are shown in Figure 2, the results of measuring 5AR2 protein expression levels are shown in Figure 3, and the results of measuring PSA protein expression levels are shown in Figure 4. It was.

According to this, the expression levels of AR, 5AR2, and PSA proteins tended to decrease in a concentration-dependent manner upon treatment with the Centella asiatica extract of Example 1. In particular, the expression levels of 5AR2 and PSA proteins decreased sharply in the 500 μg/ml treatment group. The 1000 ㎍/㎖ treatment group showed a very high expression level inhibition ability at a slightly higher level than the finasteride drug treatment group.

[Experimental example: in vivo experiment]

The experimental animal group of 8-week-old male SD rats (body weight 200±5 g) was divided into 5 groups as summarized in Table 1 below, and the BPH-induced group was intraperitoneally administered 50 mg/kg of phenobarbital after a 1-week rest period. After anesthesia by injection, the testicles and epididymis were removed, and the surgical site was sutured using suture thread. All experiments were conducted in a sterile environment. After a 3-day surgical stabilization period, the benign prostatic hyperplasia (BPH) group was treated with testosterone dissolved in corn oil and injected subcutaneously every day at a concentration of 3 mg/kg to induce benign prostatic hyperplasia. For the Centella asiatica treatment group (BPH+CA200), the Centella asiatica extract from Example 1 was dissolved in sterilized distilled water and administered orally at 200 mg/k weekly using a sonde. The positive control group, the saw palmetto administration group (BPH+Saw), was administered 100 mg/kg of saw palmetto dissolved in sterilized distilled water, and the finasteride administration group (BPH+Fi) was orally administered 1 mg/kg using a sonde every day for 4 weeks. . After the experiment was completed, the mice were sacrificed using zoletil, blood was collected through heart puncture, and serum was separated and stored at -80°C. The prostate was separated using RNAse-free surgical instruments, stored at -80°C, and then subjected to Western blot experiment.

group population Administration substance and concentration Con 6 - BPH 6 Testosterone (3 mg/kg) BPH+CA200 6 Centella asiatica extract (200 mg/kg) BPH+Saw 6 Saw palmetto (100 mg/kg) BPH+Fi 6 Finasteride (1 mg/kg)

Experimental Example 2: Measurement of prostate size and epithelial cell thickness

The results of comparing the photographs of the prostate and the weight of the prostate in each separated experimental group are shown in Figure 5, and the results of prostate index measurement are shown in Figure 6.

According to this, it was confirmed that the size of the prostate in the benign prostatic hyperplasia (BPH) group was significantly increased compared to the control group. On the other hand, the size (weight) and prostate index of the Centella asiatica administration group (BPH+CA200) were significantly reduced compared to the benign prostatic hyperplasia-induced group (BPH).

Meanwhile, microscopic pictures of prostate epithelial cells and the results of measuring epithelial cell thickness in each experimental group are shown in Figure 7.

According to this, it was confirmed that the prostate hyperplasia-induced group (BPH) had a significantly increased prostate epithelial cell thickness compared to the control group. On the other hand, it was confirmed that the epithelial cell thickness of the prostate was significantly reduced in the Centella asiatica administration group (BPH+CA200) compared to the benign prostatic hyperplasia-induced group (BPH).

*

Experimental Example 3: Analysis of ability to suppress expression of androgen signaling-related factors

Western blot experiments detailing protein expression of androgen signaling-related factors SRC-1 (steroid receptor coactivator), AR (androgen receptor), 5AR2 (5alpha reductase), and PSA (prostate specific antigen) for the above animal test groups. The analysis results are shown in Figure 8.

According to this, the expression of androgen signaling-related factors in the Centella asiatica treatment group (BPH+CA200) showed a significant overall decrease compared to the benign prostatic hyperplasia-induced group (BPH), and in particular, the amount of AR expression was significantly reduced.

Experimental Example 4: Analysis of growth factor expression inhibition ability

According to British journal of cancer, 101(12), 1949-1956, epidermal growth factor is known to play a role in preventing androgen control in prostate cancer. For reference, a representative diagram of the above paper is provided. Shown in 9. Due to androgen signaling, growth factor transcription occurs in the ARE (androgen response element), and the expression level tends to increase. Growth factors can be seen as a major factor that induces benign prostatic hyperplasia.

Accordingly, in this experimental example, to determine whether administration of Centella asiatica extract reduces the expression of growth factors in the prostate of mice that induced benign prostatic hyperplasia, a Western blot experiment was performed on the experimental groups as described above, and the results are shown in Figure 10 shown in According to this, benign prostatic hyperplasia was caused by growth factors such as EGF (Epidermal growth factor), IGF-1 (Insulin like growth factor I), TGF-1β (Transforming growth factor beta 1), and VEGF (Vascular endothelial growth factor) in the Centella asiatica group (BPH+CA200). It was confirmed that the expression level of (growth factor) was significantly decreased compared to the benign prostatic hyperplasia-induced group (BPH).

Experimental Example 5: Analysis of the ability to suppress expression of factors related to the PI3K/Akt and HIF-1 signaling pathways

Previously, in Experimental Example 4, it was confirmed that the group administered with Centella asiatica (BPH+CA200) suppressed androgen signaling, thereby suppressing the expression of epidermal growth factor (EGF). Meanwhile, factors related to the PI3K/Akt and HIF-1 signaling pathways, such as PI3K (phosphatidylinositol-3-kinase), Akt (protein kinase B), PTEN (phosphatase and tensin homologue), and HIF-1 (Hypoxia- To confirm whether the expression of inducible factors was suppressed, a Western blot experiment was performed as described above. For reference, a schematic diagram of the PI3K/Akt and HIF-1 signaling pathways is shown in Figure 11, and the results of the Western blot experiment are shown in Figure 12.

According to this, among the factors related to the PI3K/Akt and HIF-1 signaling pathways induced by epidermal growth factor (EGF) in the Centella asiatica treatment group (BPH+CA200), the expression of all related factor proteins except PTEN, a factor that inhibits PI3K, was expressed. This was found to be high.

Experimental Example 6: Analysis of the ability to suppress expression of factors related to cell proliferation in prostate tissue

The expression of PCNA (Proliferating Cell Nuclear Antigen) and Cyclin D1 (Cyclin D1) proteins, which are representative factors related to cell proliferation in prostate tissue, were analyzed by the Western blot experiment described above, and the results are shown in Figure 13. According to this, it was analyzed. It was confirmed that PCNA and Cyclin D1 protein expression was decreased in the Centella asiatica administration group (BPH+CA200) compared to the benign prostatic hyperplasia-induced group (BPH).

Although the embodiments of the present invention have been described above, those skilled in the art can add, change, delete or add components without departing from the spirit of the present invention as set forth in the patent claims. The present invention may be modified and changed in various ways, and this will also be included within the scope of rights of the present invention.

Claims (3)

Contains Centella asiatica extract as an active ingredient,
The Centella asiatica extract is a hot water extract,
A reagent composition for inhibiting the expression of at least one selected from AR (androgen receptor) and 5AR2 (5alpha reductase), factors related to androgen signaling, in LNCaP cells under in vitro conditions. delete delete