KR20060014686A - An erection accelerating agent developed from eclipta(eclipta prostrata l) a preparation method thereof and food - Google Patents

Abstract

The present invention relates to an erectile accelerator, a preparation method thereof, and a fortified food product based on extracts of nasturtium.

Nasturtium ( Eclipta prostrata L.) Extract based on ethanol / water-butanol extract, red ginseng ( Panax ginseng : red ginseng), bokbunja ( Rubus coreanus ) of silkworm pupa, whose erection accelerating activity was proved through this experiment. Extracts were added to appropriate excipients in a weight ratio of 6.0: 2.0: 1.0: 1.0, respectively, and mixed, and an appropriate amount of arginine, L-glutamic acid, tocopherol, and taurine were added as a strengthening agent to prepare a functional erection accelerating agent (EAA). It was.

DPH, erectile promoter, erectile stimulating component, cAMP degrading enzyme.

Description

An erection accelerating agent developed from Eclipta (Eclipta prostrata L) a preparation method about and erectile accelerators

The present invention is particularly hanryeoncho (Eclipta than hanryeoncho to erect the promoter presided over and extract its preparation and on his health supplements prostrata L.) Ethanol extract, and silkworm male pupae extract, red ginseng ( Panax ), whose erectile stimulation has been clarified among 82 natural foods and herbal ingredients. ginseng : red ginseng), Bokbunja ( Rubus coreanus ) extract is added as a function enhancer to a functional erectile accelerator.

Until now, the treatment of erectile dysfunction has been mainly performed by 1) vasodilator injection and 2) implant surgery. Recently, 3) intraurethral vasodilator injection and 4) oral erectile dysfunction treatment have emerged as new treatments. Among them, drug use is the most preferred method. Ideal oral medications for the treatment of erectile dysfunction should trigger effective erections as conditions and ensure their safety even when taken for long periods. However, there are no drugs that meet these conditions yet. In Viagra, the erection is excellent, but the side effects can be severe.

From 1999 to 2001, the Rural Development Administration was devoted to extracting silkworm moth and silkworm chrysalis extracts to measure NO as an stimulating factor through in vitro experiments and to measure cGMP as an erection accelerating component (EAC). Testosterone was measured as a male hormone. In addition, sperm counts were measured through in vivo experiments, and endurance tests through swimming experiments were conducted to evaluate the activity of phosphodiesterase (PDE) in order to confirm the differentiation from viagra. Has been experimentally proven to have significant side effects instead of being fast-acting.

However, Nasturtium extract has very high erectile stimulatory activity and has no side effects, and effectively inhibits LDL-cholesterol and neutral lipids, inhibits the atherosclerotic index (AI), effectively inhibits adult diseases and free radicals. The fact that it inhibits the production of oxidative stress (oxidative stress) and the active oxygen scavenging enzyme activity, such as effectively inhibit the physiological aging (physiological aging) can not be overlooked the fact that it is excellent.

Therefore, it has succeeded in developing functional erectile accelerator, its preparation, and its fortified foods from Nasturtium extract.

Until now, as a natural food material "extract silkworm moth or chrysalis extract with a sedative effect, a method for preparing the same and a food and pharmaceutical composition containing the same (patent application: 013729)" and "erection of silkworm moth or chrysalis Based on the know-how obtained in the patent application for the food and pharmaceutical composition (patent application: 0028518) of a protein fragment containing 118 amino acids from 1665 to 1782 of the non-terogenin protein as a promoter, ( Eclipta prostrata L. There is no special technical problem because it is the development of functional erectile accelerator based on extract.

Thus, 216.4 g of 60% ethanol extract was obtained from 3.0 kg of nasturtium and 40.14 g of methylene chloride (CH ₂ Cl ₂ ) fraction, 13.22 g of ethyl acetate (EtOAc) fraction, and 28.34 g of buthanol (BuOH) fraction, respectively. And 134.72 g of these fractions of water (H ₂ O) were used to evaluate erectile promoter NO, erectile promoter cGMP, male hormone testosterone, cGMP degrading enzyme PDE activity, and water-butanol extract of silkworm pupae Successfully proving that it has the best erection promoting activity, the extraction solvent was decided to be water-butanol.

The present invention is 1) extract the nasturtium with 60% ethanol and freeze-dried, and then secured water-butanol extract by a systematic organic solvent method, 2) silkworm silkworm pupa as a functional erectile strengthening material compared to the water-butanol extract of nasturtium Extracts, red ginseng extracts and bokbunja extracts were added in a weight ratio of 6.0: 2.0: 1.0: 1.0 to prepare an erection accelerating agent (EAA).

1. Materials and Experimental Methods

(1) Composition of materials and prototypes

Material of the present invention is based on 0.10 to 50.0 parts by weight of water-butanol extract fractionated using ethanol extract of Hanryokcho lyophilized product, 0.01-10.0 parts by weight of silkworm pupa extract with red erectile activity as a function enhancer, red ginseng extract and 0.01 to 10.0 parts by weight of each extract of Bokbunja are added, and 0.01 to 0.50 parts by weight of arginine as the base of cGMP, 0.01 to 0.05 parts by weight of tocopherol as antioxidant, 0.01 to 0.05 parts by weight of taurine as fatigue recovery agent and ascorbic acid. A functional erectile accelerator based on a nasturtium extract was prepared by adding 0.01 to 0.05 parts by weight of binic acid to an excipient.

Functional erection promoting agent of the present invention: production of NO to facilitate erection activity (erection accelerating agent EAA) in the testis through in vitro experiments, measuring the synthesis and testosterone production of cGMP, and sperm in the epididymis through the in vivo experiment (sperm count) was measured and evaluated.

216.4 g of 60% ethanol extract was obtained from 3.0 kg of nasturtium, followed by systematic solvent extraction. 40.14 g of methylene chloride (CH ₂ Cl ₂ ) fraction, 13.22 g of ethyl acetate (EtOAc) fraction, 28.34 g of buthanol (BuOH) fraction, and These fractions of 134.72 g of water (H ₂ O) are used to assess erectile factor NO, erectile component cGMP, testosterone and cGMP degrading enzyme PDE activity.

                       Nasturtium Dried Powder (3.0kg)

                                │

                       60% EtOH Extract (216.4 g)

                                │

                    ┌ -------- 〜-┘ ---------- ┐

                    │ ┌ -------- 〚 ----------- ┐

CH ₂ Cl ₂ fraction (40.1 g) │ │

                            EtOAc fraction (13.2g) ┌ -------- 〜 ┴ ---------- ┐

                                             │ │

BuOH fraction (28.3 g) H ₂ O fraction (134.72 g)

(2) Experimental method

NO production measurement

The production of nitric oxide (NO) in the testes was measured by some modifications of Yu et al. (1994) and Miesel et al. (1996). 0.1 g of testicle fraction was homogenized with 1.0 ml of a homogeneous buffer solution (0.1 M potassium phosphate buffer, pH 7.5), and then 200 μl of the supernatant obtained by centrifugation at 600 × g for 5 minutes. 100 μl of 90munit nitrate reductase, 0.28 mM 100 μl of NADPH, 100 μl of 3.5 μm FAD, and 200 μl of 0.1 M potassium phosphate buffer (pH 7.5) were added thereto, followed by incubation at 25 ° C. for 1 hour, followed by boiling for 3 minutes to stop the reaction. Then, 700 μl of Griss reagent (2% sulfanilic acid / 5% H ₃ PO ₄ : 0.2% N- (1-naphtyl) -ethylendiamine / water. 1: 1, v / v) was added and 10 minutes at 60 ° C. After warming, the absorbance was measured at 546 nm and quantified by a standard calibration curve (nmol / mg protein) by sodium nitrate.

cGMP of Synthetic amount  Measure

The measurement of cyclic guanonsine monophosphate (cGMP) synthesized according to NO activity in the testicles was measured using Kit reagent (Amersham Pharmacia, Code No. RPA-525 cGMP [ ¹²⁵ I] assay). A small amount of ¹²⁵ I was labeled on the binding site of cGMP-specific antibody to compare the amount of cGMP with known cGMP. Culturing the testicular fraction was used in in vitro experiments in vivo experiments the content of cGMP and displayed in a smaller quantity of ¹²⁵ for the binding site of a cGMP-specific antibody I using the supernatant as a sample to homogenize the testis fractions with 6% TCA solution Was measured. The sample was placed in a glass tube (12 × 75 mm) and 25 μl of acetylation reagent (acetic anhydride: triethylamine = 1: 2, v / v) was added. After acetylation, take 100 ㎕ of each duplicate and transfer to polypropylene assay tube, add 100 ㎕ antiserum and mix well, let stand at room temperature for 1 hour, add 100 μl of ¹²⁵ I cGMP and mix and mix at 15∼18 at 2∼8 ℃. It was left for time.

As a next step, when Amerlex-M second antibody was added, 500 μl of each tube was removed and centrifuged to completely remove the supernatant.Then, the amount of cGMP synthesis (count / min) was measured by measuring the radioactivity for 60 seconds as a γ-scintillation counter. Were analyzed and evaluated.

Determination of Testosterone Production

The testosterone production in testes was measured according to the method of Aida et al. (1984). Steroid extraction of testes was homogenized with 10 times 80% ethanol and centrifuged at 1,000 ° C for 15 minutes at 4 ° C. The supernatant was aliquoted and dried completely with nitrogen gas. The above extraction process was repeated three times. 500 μl of distilled water was dissolved in each extract, and then 4 ml of ethyl ether was added and mixed well. The mixture was left at −70 ° C. for 5 minutes. . After repeating the above procedure twice, each extract was dissolved in 4 times 0.1% gel-PBS buffer solution (pH 7.5) and stored at -20 ℃ until quantification.

100 μl of testosterone, which was radiolabeled with ³ H, was added to the sample, followed by 200 μl of diluted antibody, followed by stirring at 4 ° C. for 12 hours. In order to separate the binding type and the free type of the antigenic antibody, DCC (dextran coated charcoal) was added at 250 ° C. and left at 4 ° C. for 15 minutes, followed by centrifugation at 1,000 × g for 15 minutes to take the bound supernatant. 3 ml of scintillation cocktail was added and analyzed by liquid scintillation meter (Wallac 1409, Wallac Co.). The production amount of testosterone (ng / mg protein) was quantified according to the standard calibration curve made in 8 steps by dose.

Sperm count (sperm count)

Sperm count was measured in the epididymis according to the method of Morrissey et al. (1988). The left epididymis of the mouse was removed and stored at −70 ° C. until measurement. After dissolution at room temperature, homogenization was performed by addition of 10 times HBSS solution of epididymal weight. After diluting the homogenate 40 times with HBSS solution, a certain amount was taken and sperm count (× 10 ⁶ / g epididymis) was analyzed by an inverted microscope using a hemocytometer.

2. Experimental Results and Discussion

(1) Evaluation of production of erectile promoter (NO)

Nitric oxide identified as stimulators of erection directly related to the synthesis of cGMP in the erection promoting component (nitric oxide: NO) hanryeoncho the testis homogenate from the in vitro experiments to evaluate the amount of (Eclipta prostrata L. ) After adding the functional erection accelerating agent (EAA) prototype for each dose and mixing well, incubation at 25 ° C for 1 hour, and then analyzing the production of NO in testes.

Table 1. Comparison of NO production in testes in vitro

      Development new products               Nitric oxide (NO) production amount (nmol / mg protein)        Control        10.42 ± 0.34          100.0%        EAA-0.50 11.87 ± 0.34 ^*          114.0%        EAA-1.00 12.35 ± 0.48 ^**          118.6%        EAA-2.00 12.84 ± 0.34 ^***          123.3%        EAA-4.00 13.89 ± 0.74 ^***          133.3%

EAA-0.50, 1.00, 2.00 and 4.00: Add group of red ginseng stimulator 0.50, 1.00, 2.00 and 4.00 mg to testicular homogenate

(1 hour at 25 ° C.); Significance test: ^* p <0.05 compared to control; ^** p <0.01; ^*** p <0.001.

As you can see from Table 1, water is used as a control, nasturtium ( Eclipta prostrata L. ) The production of NO in the EAA-0.50, EAA-1.00, EAA-2.00 and EAA-4.00 addition groups as the new erection accelerating agent (EAA) prototypes in Korea was 11.87 ± 0.34, 12.35 ± 0.48, 12.84 ± 0.34 and 13.89 ± 0.74 nmol / mg protein, 114.0%, 118.6%, 123.3% and 133.3%, respectively, compared to the NO production (10.42 ± 0.34 nmol / mg protein: 100.0%) of the control group, depending on the addition amount of 14-33%. The production increase effect was recognized.

Thus hanryeoncho (Eclipta prostrata L.) An extract as main and silkworm be added to pupa extract, ginseng and Rubus coreanus extracts. Prepared a result of this by evaluating the amount increases the effect of NO, promotes highly effective erection as the erection promoting factor effect of the functional erection promoting agent You can expect a role.

(2) cGMP of Synthetic amount  evaluation

Since the NO to the body of arginine (arginine) to the substrate to promote the guanylyl cyclase activity and to the enzyme promotes the synthesis of cGMP erection promoting component in testis homogenate from the in vitro experiments to evaluate the synthesis amount of the cGMP hanryeoncho ( Eclipta prostrata L. ) Add a functional erectile accelerator (EAA) prototype by dose and mix well before incubation at 25 ° C for 1 hour, and then evaluate the amount of cGMP in the testes.

As you can see in Table 2, water is used as a control, nasturtium ( Eclipta prostrata L. As a new erection accelerating agent (EAA) prototype, the amount of cGMP synthesized in the EAA-0.50, EAA-1.00, EAA-2.00 and EAA-4.00 addition groups was 432.93 ± 13.19, 466.03 ± 24.25, 493.90 ± 9.76 and 110.8%, 119.3%, 126.4%, and 135.3%, respectively, of 528.40 ± 11.03 cpm (110.0%) compared to the control cGMP (390.60 ± 12.81 cpm: 100.0%). Was admitted.

Thus hanryeoncho (Eclipta prostrata L.) An extract as main and here silkworm can pupa, the addition of ginseng and Rubus coreanus extract. The results of this by evaluating the cGMP synthesis capacity increase effect of the thus prepared functional erection promoter, would be quite effective in the erection promoting It is expected.

Table 2. Comparison of cGMP synthesis in testicles in vitro

      Development new products                  Compound amount of cGMP (count / min)        Control        390.60 ± 12.81          100.0%        EAA-0.50 432.93 ± 13.19 ^*          110.8%        EAA-1.00 466.03 ± 24.25 ^**          119.3%        EAA-2.00 493.90 ± 9.76 ^***          126.4%        EAA-4.00 528.40 ± 11.03 ^***          135.3%

EAA-0.50, 1.00, 2.00 and 4.00: Add group of red ginseng stimulator 0.50, 1.00, 2.00 and 4.00 mg to testicular homogenate

(1 hour at 25 ° C.); Significance test: ^* p <0.05 compared to control; ^** p <0.01; ^*** p <0.001.

(3) Testosterone of Synthetic amount  evaluation

Hanryeoncho the testis homogenate from the in vitro experiments to as male testosterone representing the force to evaluate the production of testosterone (testosterone) (Eclipta prostrata L. ) Add the functional erection accelerating agent (EAA) prototyping agent and mix well, incubate at 25 ℃ for 1 hour, and then analyze the testosterone content in testes.

As you can see in Table 3, water was used as a control, nasturtium ( Eclipta prostrata L. Testosterone production of EAA-0.50, EAA-1.00, EAA-2.00 and EAA-4.00 added groups was 11.32 ± 0.50, 11.70 ± 0.52, 12.66 ± 0.09 and 13.55 as a new erection accelerating agent (EAA) prototype. ± 0.67 ng / mg testis, which is significantly dependent on the amount of EAA added of 10-32%, as 110.0%, 113.6%, 123.0%, and 131.7%, respectively, compared to the production of testosterone (10.30 ± 0.94 ng / mg testis: 100.0%). The effect of increasing testosterone production was recognized.

Thus hanryeoncho (Eclipta prostrata L.) An extract as main and can silkworm herein pupa, the addition of ginseng and bokbunja. Prepared a result of this by evaluating the amount increasing effect of testosterone in the new functional erection promoters, considerably erection promoting during long-term use It is expected to be effective.

Table 3. Comparison of testosterone production in testicles in vitro

      Development new products               Testosterone Production (ng / mg testis)        Control        10.30 ± 0.94          100.0%        EAA-0.50 11.32 ± 0.50 ^*          110.0%        EAA-1.00 11.70 ± 0.52 ^**          113.6%        EAA-2.00 12.66 ± 0.09 ^***          123.0%        EAA-4.00 13.55 ± 0.67 ^***          131.7%

EAA-0.50, 1.00, 2.00 and 4.00: Add group of red ginseng stimulator 0.50, 1.00, 2.00 and 4.00 mg to testicular homogenate

(1 hour at 25 ° C.); Significance test: ^* p <0.05 compared to control; ^** p <0.01; ^*** p <0.001.

Based on the above experimental results, administration of the functional erectile accelerator prototype of Nasturtium herbaceous acid not only promotes the production of nitric oxide (NO) as an erection accelerating factor but also promotes the stimulation of cyclic guanosine 5'-monophosphate ( It promotes the synthesis of cGMP) and the production of testosterone as a male hormone, so it is expected to be very effective in promoting erection during long-term use.

Example  One

The material of the present invention is based on 18.0 parts by weight of water-butanol extract, fractionated using ethanol extract of lyophilized product of Hanryokcho, and 6.0 parts by weight of silkworm pupa extract, red ginseng extract and bokbunja extract, each of which has an stimulating activity as a function enhancer. 3.0 parts by weight were added, and 1.0 parts by weight of arginine as cGMP base material, 0.05 parts by weight of tocopherol as antioxidant, 0.2 parts by weight of taurine and 0.05 parts by weight of ascorbic acid as excipients were added to the excipient. A new functional erectile accelerator having enhanced functionality was prepared, and compared with the prepared by weight parts of each component shown in Table 4 below. As a result of the experiment, the prepared weight parts were the most effective.

 Table 4. Prescription Ingredients and Addition of Erection Promoters (EAA) in Hannyeoncho Extract

  Material / weight part  0.05  0.20  0.50  1.00  3.00  6.00  9.00 12.00 15.00 18.00  24.00  Nasturtium Extract  ●  ●   ●  ●   ●  Chrysalis  ●  ●   ●  ●   ●  Red Ginseng Extract  ●  ●   ●  ●  ●  Bokbunja Extract  ●   ●  ●  ●  ●  Arginine  ●  ●  ●  ●  ●  Taurine  ●  ●  ●  ●  ●  Tocopherol  ●  ●  ●  ●  ●  Ascorbic acid  ●  ●  ●  ●  ●

●: Actual prescription ingredients and amount

Hanryeoncho (Eclipta based on the above experiment results prostrata L.) in the presence (主材) and facilitate erection through this experiment here, the active extract (erection accelerating activity) can be proven Chrysalis silkworm (silkworm male pupae), ginseng (Panax ginseng: red ginseng) and bokbunja (Rubus coreanus ) extracts are added to the appropriate excipients at a weight ratio of 6.0: 2.0: 1.0: 1.0, respectively.The new functional erection enhancer is enhanced by adding an appropriate amount of arginine, L-glutamic acid, tocopherol and taurine as a strengthening agent. accelerating agent (EAA) was developed.

The present invention is the extract of the silkworm pupae, red ginseng and nasturtium extracts of which the erection accelerating activity is proved among 82 natural foods and herbal ingredients, in a weight ratio of 6.0: 2.0: 1.0: 1.0 It is added to a suitable excipient and mixed with nitric oxide (NO) as an erection accelerator. In addition to promoting the production of), it promotes the synthesis of cyclic guanosine 5'-monophosphate (cGMP), an erectile promoting ingredient, and the production of testosterone as a male hormone, and thus has a significant effect on promoting erection during long-term use.

Claims (3)

Water-butanol extract of Nasturtium extract with 60% ethanol, ethanol / water-butanol extract of silkworm pupa, red ginseng extract, and bokbunja extract were added to the appropriate excipients in a weight ratio of 6.0: 2.0: 1.0: 1.0, respectively. A method of producing an erection accelerator based on a nasturtium extract, characterized by adding an appropriate amount of arginine, L-glutamic acid, tocopherol, and taurine as strengthening agents. It is based on 0.10-50.0 parts by weight of Hannyeoncho water-butanol extract, and 0.01-10.0 parts by weight of silkworm pupa extract, red ginseng extract and bokbunja extract each having an erection promoting activity as a function enhancer, and added cGMP Nasturtium prepared by mixing and mixing 0.01 to 0.50 parts by weight of arginine as base, 0.01 to 0.05 parts by weight of tocopherol as antioxidant, 0.01 to 0.05 parts by weight of taurine and 0.01 to 0.05 parts by weight of ascorbic acid as fatigue recovery agent. Erectile accelerator based on extract. 18.0 parts by weight of water-butanol extract, fractionated using ethanol extract of lyophilized product of Hanryokcho, was added, and 6.0 parts by weight of silkworm pupa extract, red ginseng extract and bokbunja extract, each with erectile promoting activity, were added as a function enhancer. Here, 1.0 parts by weight of arginine as cGMP base material, 0.05 parts by weight of tocopherol as antioxidant, 0.2 parts by weight of taurine and 0.05 parts by weight of ascorbic acid as fatigue recovery agent are added to the excipient. One erection fortified food.