KR20220160301A - Composition for Prevention or Treatment of Prostatic Desease Comprising Sargassum Horneri Extract - Google Patents

Abstract

The present invention relates to a composition with excellent efficacy in the prevention or treatment/alleviation of symptoms of prostatic disease derived from a natural product, which does not cause side effects or drug resistance of conventional therapeutic agents, is low in toxicity, and can be administered for a long period of time. More specifically, the present invention relates to a composition for preventing or treating/alleviating symptoms of prostatic disease which contains a Sargassum horneri extract as an active component.

Description

Composition for Prevention or Treatment of Prostate Disease Containing an Extract of Capricorn Capitis {Composition for Prevention or Treatment of Prostatic Desease Comprising Sargassum Horneri Extract}

The present invention relates to a composition with excellent efficacy in preventing or treating/improving symptoms of prostate disease derived from natural products that does not cause side effects or drug resistance of conventional therapeutic agents and has low toxicity and can be administered for a long period of time.

The prostate is an accessory gonad composed of glandular tissue and fibromuscular tissue that produces and secretes semen and blocks urinary tract infections. The prostatic fluid produced in the prostate gland supplies nutrients to the sperm that have moved from the testis, and helps the sperm to move actively by maintaining a liquid state so that the ejaculated semen does not harden. Representative prostate-related diseases include prostatic hyperplasia, which mainly occurs after the age of 50, and prostatitis and prostate cancer, which mainly occur in the younger age group before the age of 40.

Benign prostatic hyperplasia (BPH) is a disease that causes various symptoms such as dysuria by compressing the urethra due to abnormal growth of the prostate around the urethra. In general, the size of the prostate in adults is about 20g, but when the prostate enlarges and grows to an abnormal size of 40-400g, it presses on the nearby urinary tract and causes urinary frequency of urinating more than 8 times a day, nocturia, and strong and sudden urge to urinate. If you have a urge to urinate while urinating, symptoms of bladder storage symptoms such as unbearable urgency, symptoms of bladder discharge disorders such as delayed urine in which urine comes out only when you urinate, interrupted urine in which urine flow is interrupted, and a phenomenon in which you have to apply force during urination appear. do. Although the cause of the disease has not yet been clearly identified, it is accepted that physical aging and changes in male hormones affect it. When testosterone is excessively present in the blood, a large amount of dihydrotestosterone (DHT) is synthesized by 5-alpha reductase present in prostate tissue. It binds to the androgen receptor and induces the growth of the cytoplasm of the epithelium and stroma.

According to the National Health Insurance Corporation's analysis of health insurance big data for the past six years (2012-2017), as of 2017, the number of people who received treatment for benign prostatic hyperplasia accounted for 5.1% of the total number of health insurance patients, which is one of the most common diseases among adult males. One. The recent outbreak has been rapidly increasing, with an average annual increase rate of 5.9% over the past six years. Prostatic hyperplasia is commonly said to be 50% in the 50s, 60% in the 60s, 70% in the 70s, and 80% in the 80s. It is also an increasing trend.

Patients with prostatic hyperplasia do not require surgical treatment unless there is urinary tract obstruction or other complications, but about 50% of patients up to the age of 80 require treatment. Prostatic hyperplasia is not a fatal disease in itself, but it deteriorates the quality of life. In addition, if left untreated without appropriate treatment, it can cause various stone diseases in the kidneys, cause urinary tract infection due to persistent residual urine in the bladder, and in severe cases, urinary sepsis, which can affect life support. In addition, in that some patients with BPH are related to prostate cancer, it has emerged as an important medical problem in the current society entering a super-aged society.

The treatment method for benign prostatic hyperplasia was performed mostly through surgery in the early days, but since the late 1980s, it has rapidly changed to a drug administration method. Alpha-adrenergic antagonists such as terazosin, doxazosin, and tamsulosin improve symptoms immediately after taking them, but they are drugs for symptom improvement rather than treatment. 5-α reductase inhibitors such as finasteride and dutasteride need to be taken for at least 6 months to see the response, and reduce libido and sexual function, and when administered to patients with high-grade prostate cancer There is a problem with increasing risk. As such, drugs used in the treatment of conventional prostate diseases exhibit many side effects and have limitations in their efficacy. Recently, various drugs extracted from natural products have been developed and used, but they have not yet been widely recognized. Therefore, it is necessary to develop a new therapeutic agent having excellent therapeutic effect without causing side effects or drug resistance.

Prostatitis is a disease in which the prostate is inflamed, and it is a disease so common that about 50% of adult men experience symptoms at least once during their lifetime. According to the statistics of the Health Insurance Review and Assessment Service, 260,000 people in 2016 and 270,000 people in 2019 are constantly increasing to the extent that they visit hospitals for prostatitis. Bacterial prostatitis caused by infection accounts for only 5-10% of prostatitis, and chronic non-bacterial prostatitis accounts for most of it. Bacterial prostatitis is most often caused by direct infection of bacteria through the urethra during urinary tract infections, and in addition, it can be caused by impaired excretion of prostatic fluid, urinary reflux into the prostate, and transmission of inflammation such as hemorrhoids or colitis. . Chronic nonbacterial prostatitis is considered to be caused by improper lifestyle habits such as excessive drinking, smoking, overwork, and stress, or continuous stimulation to the prostate, such as sitting for a long time or riding a bicycle.

Prostate cancer is the most common male cancer in Western countries and shows a high incidence, and its frequency is rapidly increasing in Korea as well. The treatment of prostate cancer differs depending on the stage of disease progression. In the case of local cancer, treatment is aimed at fundamental treatment, but in the case of metastatic cancer, systemic treatment is performed. For the treatment of localized prostate cancer, waiting therapy, radical prostatectomy, and radiation therapy can be selected in consideration of the patient's age, health status, sexual function status, tumor stage and differentiation, and patient preference. Metastatic prostate cancer can be treated with hormone therapy or chemotherapy.

Since prostatitis or prostate cancer as described above is also related to androgen, which is a male hormone, anti-androgen drugs that inhibit the expression of androgen receptors can treat and prevent diseases or relieve symptoms.

The hoesaeng hatban is a perennial brown algae with strong fertility. In particular, the inflow to Jeju accounts for 90% of the total, and its roots are buried deep in the sea, making it difficult to remove them. As a result, they are regarded as uninvited guests of the sea that cause considerable damage, such as damage to natural scenery, generation of odor, loss of tourism industry, damage to the overall fishery due to disruption of ship operation and navigation, and enormous manpower and budget for control. It is known to have various physiological functions and pharmacological effects because it contains various components such as fucoidan and alginic acid, minerals, and polyphenols. In addition, it is disclosed that hoeeren cap is useful for preventing or treating/improving symptoms of prostate disease.

Registered Patent No. 10-1652073 Registered Patent No. 10-1733477 Registered Patent No. 10-1688018

An object of the present invention is to provide a novel composition effective for preventing and treating/improving symptoms of prostate disease in order to solve the problems of the prior art as described above.

In addition, an object of the present invention is to provide a composition for preventing and treating/improving symptoms of prostate disease that is non-toxic and can be safely used without side effects or drug resistance.

Another object of the present invention is to provide a new way to utilize the hoesaeng cap, which is emerging as an environmental problem as an uninvited guest in the sea.

In order to achieve the above object, the present invention relates to a pharmaceutical composition for preventing or treating prostate diseases selected from the group consisting of prostate cancer, prostatic hyperplasia and prostatitis, characterized in that it contains an extract of chinensis chinensis as an active ingredient.

In the measurement of expression of androgen receptor (AR) and prostate-specific antigen (PSA) using LNCaP cells, a prostate cancer cell line commonly used as an experimental model for prostate disease, the oxen cap extract effectively inhibited the expression of AR and PSA in a dose-dependent manner. It was shown to be effective in preventing and treating prostate diseases by suppressing In particular, the extract of A. japonica extract was more effective in suppressing the expression of AR, and showed an excellent effect among other algae of the genus Mosquito genus.

In addition, in an animal model in which prostatic hyperplasia was induced by administration of testosterone, the composition containing the oleander capsular extract of the present invention effectively reduced the size and weight of the enlarged prostate. In detail, the extract of spp. extract effectively suppressed the increase in the thickness and number of thickened prostate epithelial cells by reducing the blood concentration of dihydrotosterone, which induces the cytoplasmic growth of the epithelial and stroma of the prostate.

In the present invention, the prostate disease is a disease affected by male hormones and may be prostate cancer, benign prostatic hyperplasia or prostatitis.

The extract of the black capsicum of the present invention includes all forms in which useful components of the black capsicum are concentrated. Most simply, the oxen capsular extract may be a dried product or dry powder of hoemens caps from which moisture has been removed. Alternatively, it can be prepared by extracting living organisms or dried products of C. chinensis using one or a mixture of two or more of C1-C4 lower alcohol and water as a solvent. Extraction may be performed by simply immersing in a solvent at room temperature, but extraction may be performed by using ultrasonic waves or by heating and stirring as in the following examples to increase extraction efficiency within a short time. Also, to make the extraction more efficient, the hoesaeng cap can be extracted by crushing or grinding into small pieces. During the extraction, the amount of the solvent may be used in a volume ratio of 2 to 50 times the weight of the hoesaeng cap. Since the extraction efficiency increases as the amount of the solvent increases, it is not a problem to use more than 50 times the solvent, but considering the efficiency and economics of the process, the amount of the solvent is preferably 50 times or less. In addition, in order to increase the extraction efficiency while using the same amount of solvent, it is recommended to increase the number of extractions.

The present invention also relates to a pharmaceutical composition for improving dysuria due to benign prostatic hyperplasia, characterized in that it contains an extract of capsular capillaries as an active ingredient. The composition of the present invention can effectively improve dysuria because it relieves pressure on the urethra by effectively reducing the size of the thickened prostate.

Dosage of the pharmaceutical composition containing the extract of the black capsular capillaries of the present invention as an active ingredient will vary depending on the age and weight of the subject to be treated, the severity of the specific disease or pathological condition to be treated, the route of administration, and the judgment of the prescriber. The composition of the present invention can be prepared by a method known in the pharmaceutical field for use as a drug for treatment, and can be used by itself or in combination with a pharmaceutically acceptable carrier, excipient, diluent, etc. can The composition of the present invention can be formulated into a preparation for oral or parenteral administration and used as an agent for the treatment and prevention of prostate diseases. In addition, the composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and when administered in combination with other therapeutic agents, it may be administered sequentially or simultaneously. The dosage of the active ingredient according to the present invention can be determined by those skilled in the art so that the maximum effect can be obtained with the minimum amount without side effects in consideration of the absorption of the active ingredient in the body, the form of the preparation, the age and condition of the patient, the degree of symptoms, etc. It can be appropriately selected by, and administration may be administered once a day or divided into several times. A typical dosage is 0.001 mg/kg·day to 10 g/kg·day.

In addition, the present invention provides a health functional food composition for preventing and improving symptoms of prostate disease, comprising an extract of thorax chinensis as an active ingredient. The extract of the thoracic capitis may be added to the health functional food of the present invention in an amount of preferably 0.01 to 100% by weight. The health functional food of the present invention includes forms such as tablets, capsules, pills or liquids, and foods to which the composition of the present invention can be added include, for example, various foods, beverages, gum, tea, and vitamin complexes. , health functional foods, etc.

As described above, according to the present invention, by inhibiting the expression of androgen receptor and prostate-specific antigen, it can be effectively used for prevention and treatment/improvement of symptoms of prostate disease.

In addition, the composition of the present invention is made of natural materials and can be safely used without worrying about toxicity or side effects, and it provides a use that can be used as a pharmaceutical or health functional food composition with high added value of the hoesaeng hat, which is emerging as an environmental problem, to protect the environment. At the same time as solving the problem, it can provide a new source of income for fishermen.

1 is an image and a graph of Western blot results showing that an extract of the chinensis cap in LNCap cells exhibits an inhibitory effect on AR and PSA expression.
Figure 2 is a Western blot result image and graph showing the AR expression inhibitory effect of extracts from the genus Mosquito in LNCap cells.
Figure 3 is a photograph showing the effect of administration of the hoesaeng capban extract on the size of the prostate in an animal model of benign prostatic hyperplasia.
Figure 4 is a graph showing the effect of administration of the hoesaeng capban extract on body weight and prostate weight in an animal model of benign prostatic hyperplasia.
Figure 5 is a graph showing the effect of the administration of hoesaeng capban extract on blood DHT concentration in an animal model of benign prostatic hyperplasia.
Figure 6 is a microscope image showing the effect of the administration of the hoesaeng capban extract on the thickness of prostate epithelial cells in an animal model of benign prostatic hyperplasia.
7 is a graph showing the effect of the administration of the black capsular extract on the thickness of prostate epithelial cells in an animal model of benign prostatic hyperplasia.

Hereinafter, the present invention will be described in more detail with reference to the attached examples. However, these embodiments are only examples for easily explaining the content and scope of the technical idea of the present invention, and thereby the technical scope of the present invention is not limited or changed. It will be obvious to those skilled in the art that various modifications and changes are possible within the scope of the technical spirit of the present invention based on these examples.

[Example]

Example 1: Preparation of extracts of oyster capsicum

The hoesaeng hat half was dried and used from the coast of Jeju Island in March 2020. 2.5 L of 30% ethanol was added to 500 g of dried hoesaeng caps, and extraction was performed at room temperature for 2 days while stirring. When the extraction was completed, solids were removed by filtration, and the filtrate was concentrated under reduced pressure to obtain 28.32 g of an extract. The extract was stored frozen and used.

Example 2: Evaluation of the inhibitory effect of PSA and AR expression of hoeer cap in LNCaP cells

The following experiment confirmed the effect of the extract on the expression of prostate-specific antigen and androgen receptor.

LNCaP cells (ATCC, USA), a male hormone-dependent human prostate cancer cell line, were plated in a 6-well plate at a concentration of 5×10 ⁵ cell/well and supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptoprotein. It was cultured at 37°C for 15 hours in medium containing mycin. The cultured cells were treated with 1 nM of testosterone (TP, Testosterone Propionate; TCI, Japan) and the extract (S26) at a concentration of 10, 50 or 100 μg/ml, and cultured for additional 72 hours, and then harvested. As a negative control group, only 1 nM of testosterone was treated without treatment of the genus chinensis extract, and as a positive control group, 10 μM of finasteride (Fina, Finasteride), a representative prostatic hyperplasia treatment, was treated with 100 nM of testosterone instead of the chinensis chinensis extract. The untreated group was referred to as the normal group (Control).

Cells harvested after culture are treated with RIPA buffer (50 mM Tris-HCl, pH 8.0, with 150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail) and centrifuged at 12,000 rpm for 20 minutes to separate proteins. The separated protein was quantified by the Bicinchoninic acid (BCA) method.

30 μg of the quantified protein was processed on SDS-PEGE gel, electrophoresis was performed, and the protein was transferred to a PVDF membrane. Then, the PVDF membrane was reacted with 5% skim milk for about 1 hour to remove non-specific binding proteins on the surface, and the primary antibodies AR, PSA, and β-actin were diluted 1:1000, 1:1000, and 1:5000, respectively. and reacted at 4°C for one day. After that, the secondary antibody (Anti-goat; 1: 5000, Santacruz, CA, USA, Anti-Rabbit, 1:5000, Abfrontier, Seoul, Korea) was treated at room temperature for 1 hour, and AR and PSA using ECL kit The protein expression of was confirmed.

1 is an electrophoresis image showing the results of Western blotting for PSA and AR proteins, and a graph showing the expression levels of PSA and AR proteins in each experimental group. In FIG. 1 , it can be seen that the capsular capitol extract inhibits both the expressions of PSA and AR in a dose-dependent manner, and is particularly effective for the expression of AR.

Example 3: Comparison of AR expression inhibition ability of capsular algae

Other caps other than capsular caps were dried, and extracts were prepared in the same manner as in Example 1.

According to the method of Example 2, when each extract was treated at a concentration of 100 μg/ml in the culture medium, the AR expression inhibitory effect was evaluated.

Fig. 2 is a graph showing the results. In the case of the black cap, the AR expression inhibitory effect was the best, showing more than twice the inhibitory effect than the solitary cap (8) or the small prickly cap (16), and it was the only control group among the experimental groups. It showed better effects than phosphorus finasteride.

Example 4: Efficacy Evaluation of Mosquito Mantis Extract in Animal Models

Efficacy was evaluated in an animal model of benign prostatic hyperplasia using the extract prepared in Example 1. As experimental animals, 42 male Sprague-Dawely rats (6-week old upon receipt, Orient Bio, Korea) were used in the experiment after a 7-day acclimatization process. During the acclimatization process and throughout the experiment, the rats were accommodated in a polycarbonate breeding box in a breeding room where the temperature (20-25 ° C) and relative humidity (30-35%) were controlled, and the light:dark cycle was 12 hours. The cycle was controlled, and feed (Orient Bio, Korea) and drinking water were freely supplied. Experimental animals used in this experiment were handled according to the Guide for the Care and Use of Laboratory Animals [Department of Health, Education, and Welfare Publication (National Institute of Health) 85-23].

Experimental animals were divided into 6 groups of 7 animals per group so that the average weight was constant, as shown in Table 1 below.

Testosterone (TP, Testosterone Propionate; TCI, Japan) was dissolved in corn oil at a concentration of 5 mg/ml, and then a 5 mg/kg dose was subcutaneously injected once daily for 28 days with a 1ml syringe with a 27 gauge needle attached. . In the normal control group (NC), only corn oil was injected in the same way.

Finasteride or oxen cap extract was prepared by preparing a solution in which the doses listed in Table 1 above were dissolved in 5 ml of PBS (Phosphate buffer saline), and then taken orally by force using a syringe attached with a metal Zonde once daily for 28 days. administered. Only PBS was orally administered to the normal control group and the negative control group in the same manner.

1 day before the start of drug administration, 7 days after administration, 14 days and 28 days after administration, in order to minimize weight change according to feed intake, fasting was performed for about 18 hours, and the body weights of all experimental animals were measured. 28 days after the start of drug administration, the body weight was measured and the prostate was removed by sacrificing the experimental animals. Fat and foreign substances around the extracted prostate were thoroughly removed, and the weight was measured.

Table 2 below shows the body weight, prostate weight, and prostate weight per body weight before sacrifice for each experimental animal group, and FIG. 4 shows the data graphically.

No significant difference in body weight was observed between each experimental animal during the experimental period. Testosterone injection induces prostatic hyperplasia, showing that the prostate weight per body weight increased more than twice in the benign control group compared to the normal control group. The group administered with the oxen capricorn extract exhibited an effect of reducing the prostate weight per body weight to a degree similar to that of finasteride.

5-α reductase converts testosterone into an active form of dihydrotestosterone (DHT), and DHT binds to androgen receptors to enter the nucleus and promotes the expression of genes related to prostate disease. Accordingly, the concentration of DHT in serum was analyzed by collecting blood from tail arterial blood before sacrifice of the experimental animals, and the results are shown in FIG. 5 .

In the BPH experimental model in FIG. 5, the level of DHT in serum was significantly increased compared to the control group. The DHT level in the 50 mg/ml administration group was decreased compared to the negative control group, but it did not show significance based on p<0.05. In contrast, the 100 mg/ml administration group and the 200 mg/ml administration group showed a significant (p<0.05) decrease in blood DHT. In particular, the 100 mg/ml administration group had a blood DHT concentration similar to that of the positive control finasteride administration group.

Figure 6 is an image observed under a microscope (X400) after H&E staining of prostate tissue, and Figure 7 is a graph showing the thickness of epithelial cells from the image in terms of numbers. Referring to Figures 6 and 7, it can be confirmed that the epithelial thickness of the prostate increased by the induction of prostatic hyperplasia.

Administration of the oxen cap extract effectively reduced the thickness of the prostate epithelium, which was increased by the induction of benign prostatic hyperplasia, and the effect was similar to or better than that of the positive control group.

Claims (7)

A pharmaceutical composition for preventing or treating a prostate disease selected from the group consisting of prostate cancer, benign prostatic hyperplasia and prostatitis, characterized in that it contains an extract of black capped cape as an active ingredient.
The method of claim 1,
The pharmaceutical composition according to claim 1, wherein the extract of thoracic thorax is a dried product or dry powder of thoracic thorax.
The method of claim 1,
The pharmaceutical composition according to claim 1 , characterized in that the extract of the black cap cap is extracted using one or a mixture of two or more selected from the group consisting of water and lower alcohols of C1 to C4 as a solvent.
The method according to any one of claims 1 to 3,
The pharmaceutical composition, characterized in that the extract of the black capsular capillaries inhibits the expression of androgen receptors or prostate-specific antigen proteins.
The method according to any one of claims 1 to 3,
The pharmaceutical composition according to claim 1, wherein the extract of the thoracic thorax reduces the content of dihydrotestosterone in the blood.
A pharmaceutical composition for improving urination disorders caused by benign prostatic hyperplasia, characterized in that it contains an extract of genital warts as an active ingredient.
A functional health food composition for preventing or improving symptoms of a prostate disease selected from the group consisting of prostate cancer, prostatic hyperplasia and prostatitis, characterized in that it contains an extract of the black capped cape as an active ingredient.

Images