KR20020025637A - A extract derived from male pupae and adults of silkworm, a preparation method thereof and foods and pharmaceutical compositions containing the same - Google Patents

Abstract

PURPOSE: Provided is an extract derived from male pupae and adults of silkworm. Also, its preparation method and foods and pharmaceutical compositions containing it. The extract contains active ingredient for treating impotency, and is thus used as a material for manufacturing medicines. CONSTITUTION: The method for manufacturing the extract of male pupae and adults of silkworm is characterized by selecting male pupae and adults of silkworm using a silkworm kind of which gender can be distinguishable by colors, patterns or the like. The pharmaceutical composition characteristically contains an effective amount of the extract.

Description

Silkworm moth or chrysalis extract with sedative effect, preparation method thereof and food and pharmaceutical composition containing the same {A extract derived from male pupae and adults of silkworm, a preparation method about and foods and pharmaceutical compositions containing the same}

It is an object of the present invention to provide a caustic agent derived from a natural active substance having an excellent caustic effect by using silkworm moths.

Recent economic growth places more emphasis on overall quality of life than on economic margins. Silkworms, which have contributed warmly to humanity by providing high quality fibers called silk to our humans for the past 4-5 thousand years, are now contributing greatly as a new type of natural active material production tool. In the age of treating diseases while neglecting the quality of life by synthetic medicines, there is a need for means to cure diseases while ensuring quality of life using natural active substances. In consideration of the fact that silkworm moths and pupa have been used as a suitable material, if scientific development is made, it will contribute greatly to the health of the people, the domestic sheep farming farms, and the world's sheep farming industry.

Recently, due to the high economic growth, the frequency of erectile dysfunction of organic causes is increasing due to the aging of the population and the increase of the elderly population as well as the increase of adult diseases. However, due to the tendency to pursue a better quality of life due to the improvement of economic power, the demand for treatment and improvement of this is increasing year by year.

Until now, the treatment of erectile dysfunction has been predominantly the treatment such as vasodilator injection and implant surgery, but recently, urethral vasodilator injection and oral erectile dysfunction treatment have become a new treatment.

Among them, I prefer the most effective medications. This is evident in the hot interest of the recently developed Viagra. Ideal oral medications for the treatment of erectile dysfunction should induce effective erections and their safety should be ensured even for long-term use. However, there is no effective drug that satisfies all these conditions so far.

From ancient times, silkworm moths (one-sleeping) have been found to be in agreement with Bosim and others. "Strengthen Yangyang, stop setting and bleeding, warm water, add regularity, It is recorded that fatigue does not come even after hardening, but it is not yet clear how the jeongjeong effect is manifested. It is not.

The life history of silkworms is as follows. Silkworms are insects that completely transform, and their first generation overwinters as eggs after four periods of egg, larva, pupa and moth. Silkworm eggs hatch after a certain period of time to become ant-nue, and ant-nude to sleep after eating mulberry leaves for about 3-4 days. As such, the first state of sleep is called 1 side, and the silkworm at this time is called 1 side sleep. On the first side, sleep for about 16 hours, take off the fault, and become 2nd age silkworm. Silkworms usually sleep 4 times and are 4 sleep. Silkworms are 5 years old when they eat mulberry leaves for about 7-8 days and grow the largest. The silkworm at this time is called ripe silkworm. Ripe silkworms are made in cocoons for 2-3 days, and then into pupa, after which they are transformed into moths after 10-15 days, break through silkworm cocoons, spawn mating and finish the first generation.

As a result of focusing on the necessity of natural substance-derived scavenger and the silkworm scavenger effect, the present inventors have maximized from silkworm moths and pupa which do not copulate the sedative effect substance derived from natural substance as a scavenger for taking. It can be obtained, by using this to effectively screen uncopied male moth and pupa, and to use the hardening effect of the extract of the selected male pupa and male moth extract pharmaceutical composition and / or pharmaceutical composition having a hardening effect To invent.

Figure 1 shows a picture of a Han Hwang dog can distinguish male and female by the color of the cocoon.

Figure 2 shows a picture of Hanseongban can be distinguished male and female with or without the pattern of the cocoon.

Figure 3 shows a picture of silkworm male moth and pupa.

4 is a graph illustrating endurance measurement results by forced swimming time. 1 is a control group (physiological saline), 2 is a 200 mg / kg group of male moth extract, 3 is a 200 mg / kg group of male pupa extract, 4 is a group of 200 mg / kg silkworm, 5 is a group of 50 mg / kg viagra, 6 is a tocopherol group 100 mg / kg administration group. Significance test P <0.5.

The present invention comprises the steps of screening male pupa and male moth by separating male and female pupa using silkworm varieties that can be distinguished by male or female silkworm moth and male pupa by the color of cocoon or by the presence or absence of a pattern of silkworm larvae. It relates to a method for producing an extract of uncoated male moth or pupa having a firming effect.

The present invention also relates to a freeze-dried extract of pupa or pupa moth collected by the above production method.

The present invention is a food and active ingredient having a jeongjeong effect characterized in that it comprises the hydrangea extract or pupae extract, characterized in that it comprises the hydrangea extract or pupae extract in an amount effective to exhibit a jeongjeong effect It relates to a pharmaceutical composition.

First of all, in preparing extracts of silkworm moths and chrysalis to be used for enhancement of erectile dysfunction and masculinity, it is necessary to select uncoated male moths and chrysalis. These uncoated male moths and pupa can be selected as follows.

Specifically, the screening method of uncoated male moth and pupa of the present invention can distinguish male and female by the color of cocoon (Hwang: ♀, white: ♂), such as the Hanswang silkworm breed, or the silkworm larvae, such as the Korean pattern silkworm breed. Using silkworm varieties that can be distinguished by male and female pattern by the pattern of (반: ♀, civil ♂: ♂), when distinguishing the color of cocoons, white cocoons (♂) and yellow cocoons (♀) are separated in the cocoon stage. When mating is not possible and the pattern of silkworm larvae is distinguished, it is separated and bred by the pattern ♀ of larvae at 5th age larvae, and then immediately converted to moths from ♂ 2-4 hours before By harvesting, only male moths are screened. The most desirable stages of male moth and chrysalis are pupa, which is 2-4 hours before the silkworm pupa 10-15 days early sunrise (5-7 o'clock). What is not called male moths.

In addition, the extract from the male moth or the pupa selected by the above method is most preferably the extract from the silkworm which has been almost transformed into a moth, and the pupa selected by a series of screening operations. Nitrogen treatment is stored in an ultra-low temperature freezer, lyophilized using a lyophilizer and made into a powder state, and then 50% ethanol extracted and concentrated and freeze-dried powdered.

Specifically, the extract is treated by freezing the liquid nitrogen to the selected chrysalis or moths and stored in an ultra-cold freezer, and then powdered using a freeze dryer, followed by extraction with ultrasonic volume of 20 times 50% ethanol for about 1 hour. After repeating about 5 times by filtration with filter paper and concentrated using a rotary vacuum concentrator and then lyophilized. These extract samples were used to test the jeongjeong effect and endurance test through the following test tube and animal experiments.

The production method of the extract of the male moth or chrysalis has its characteristics in the method of sorting the male moths and chrysalis which are not mated, and other extraction processes can be used if it is a conventionally known method.

In the food comprising the male moth extract and / or pupa extract, it is possible to prepare a food having a firming effect of the present invention by combining the extract of the present invention with a food additive that can be added conventionally.

In addition, it will be possible to prepare a food having a stiffening effect by adding it to a conventional favorite food.

The food comprising the extract of the present invention relates to a beverage containing the extract of the moth or pupa of the present invention in an amount effective to exhibit a jeongjeong effect, red beet juice and DHA in water-soluble calcium, and The present invention relates to Kangjung's health supplements containing oligosaccharides, liquid fructose, citric acid, β-cyclodextrin, vitamin C and vitamin D3, cinnamon extract, ginger extract, and mixed fruit flavors.

A pharmaceutical composition comprising the male moth extract and / or the pupal extract of the present invention in an amount effective to exhibit a sedative effect, comprising one or more non-toxic, pharmaceutically acceptable carriers, adjuvants or diluents, or other active ingredients It may include.

The pharmaceutical composition of the present invention may be formulated into an oral dosage form in the form of a dry powder for use as a solution, a suspension or an emulsion in an oil or an aqueous medium, or dissolved in sterile, pyrogen-free water before use. .

The compositions of the present invention may be prepared by known methods using pharmaceutically acceptable carriers and excipients, for example tablets, troches, sugar-containing tablets, aqueous or oily suspensions, dispersible powders or particles, emulsions, soft or hard. It is formulated for oral use in the form of capsules, syrups, elixirs, which are contained in unit dosage forms and in multidose containers.

In oral formulations, tablets may be inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, sodium phosphate; Granulating agents such as corn, starch and alginic acid; Disintegrants; Binders such as starch, gelatin and acacia; The compound of the present invention is mixed with an excipient which can be used for the preparation of tablets, such as lubricants such as magnesium stearate, stearic acid and talc. Tablets may be used uncoated or coated to inhibit absorption and degradation of the tablets in the gastrointestinal tract. For example, you may apply time inhibiting substances, such as glyceryl monostearate and glyceryl distearate. Hard capsule is a compound of the present invention is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, kaolin, etc., soft capsule is a solvent such as polypropylene glycol, PEGs (polyethylene glycol), ethanol, etc. that can be mixed with water And the active ingredients in oil solvents such as peanut oil, liquid paraffin and olive oil.

The water-soluble suspending agent is a mixture of excipients and active ingredients suitable for the preparation of water-soluble suspending agents. Excipients include, for example, sodium carboxymethyl cellulose, methyl cellulose, hydroxy-propyl methyl cellulose, sodium alginate, polyvinyl-pi Suspending agents such as ralidone, gum tragacanth, gum acacia and the like; Compounds condensing fatty acids such as polyoxyethylene stearate and alkylene oxides; Compounds in which alkylene oxides are condensed with long fatty acids such as heptadecaethyleneoxycetanol; Compounds which condensate partial esters derived from anhydrous hexitol anhydride and fatty acids, such as polyoxyethylene sorbitol monooleate, with ethylene oxide; Wetting agents; Or dispersing agents. Water-soluble suspensions contain preservatives, colorants, spices, sweeteners and the like.

The oily suspension is a suspension of an active ingredient in a vegetable oil such as olive oil, sesami oil, or a mineral oil such as liquid paraffin. For example, a thickening agent such as beeswax, hardened paraffin, cetyl alcohol, etc. may be used. It contains. It also contains preservatives, colorants, spices, sweeteners and the like, which can be preserved by adding antioxidants such as vitamin C.

Dispersible powders and particles have an active ingredient in a state in which a dispersing agent, a wetting agent, a suspending agent, a preservative and the like are mixed together. Suitable dispersing, wetting or suspending agents can be mentioned by way of example already mentioned above. Additional excipients are, for example, sweeteners, spices, colorants and the like.

Water-in-oil emulsions are derived from vegetable oils such as olive oil or mineral oils such as liquid paraffin, and are derived from natural phospholipids such as soy bean lecithin, and esters of anhydrous hecitol or fatty acids such as sorbitan monooleate. The active ingredient is emulsified using compounds obtained by condensation of ethylene oxide with partial esters derived from anhydrous hexitol anhydride and fatty acids such as oxyoxysorbitol monooleate.

Syrups and elixirs are a mixture of active ingredients with sweeteners such as glycerol, propylene glycol, sorbitol and sucrose.

When using the composition of the present invention, the dosage of the active ingredient may vary depending on the age, weight, general health, sex, meal, time of administration, rate of excretion, combination of drugs, degree of disease during treatment, etc. In particular, but not limited to, preferably 100 to 500 mg daily may be used daily, based on a person weighing 60 kg.

On the other hand, the amount of the compound of the present invention mixed with the carrier material to determine one dosage form depends on the mode of administration and the patient being treated.

The silkworm moth-related products seek to maximize the effects by identifying the growth effects and preparation methods, as well as elucidating the jeongjeong effect, that is, the male hormone production and erection promotion effects, the sperm productivity and the endurance test. The silkworm extract samples used in the following experiments were frozen by treating liquid nitrogen with selected chrysalis or moths and liquidized in a cryogenic freezer, and then pulverized using a freeze dryer, and then ultrasonically added with 20 times the volume of 50% ethanol. Extraction was repeated about five times for about 1 hour, filtered through filter paper, concentrated using a rotary vacuum concentrator, and then lyophilized.

1.Testosterone content determination

Testosterone content was measured by the o-phthaldehyde method. First, the silkworm extract sample was dissolved in distilled water at a concentration of 100 mg / dL 250 mg / dL and 500 mg / dL, where 0.1 mL of each was taken and then 33% KOH solution 0.3 After adding ㎖ and 3.0 ml of 95% ethanol and mixing well, serum is heated for 15 minutes, brain cell ash for 60 minutes in a 60 ℃ water bath and cooled.

5.0 ml of nucleic acid was added thereto, mixed, 3.0 ml of distilled water was added thereto, and then mixed well for 1 minute. The layers were separated, and 1.0 ml of nucleic acid layer was taken. The nucleic acid layer was concentrated to dryness with nitrogen, and 2.0 ml of o-ftaldehyde reagent was added and mixed well. After 10 minutes, 1.0 ml of concentrated sulfuric acid was added and mixed as a color developing reagent.

Within 10 to 90 minutes after sulfuric acid addition, the absorbance was measured at 550 nm using a spectrophotometer and the total testosterone content (mg / dl serum) was quantified by a standard calibration curve.

1) testosterone content

Testosterone content, which is a male hormone, is closely related to masculinity, and Viagra and Kissometer, which are focused on the eyes, are administered as control drugs, and 100mg, 250mg, and 500mg of lyophilized silkworm extracts are administered, followed by using a spectrophotometer. Absorbance at 550 nm was measured and total testosterone content (mg / dl serum) was quantified by standard calibration curve.

As a result, as shown in Table 1, when the testosterone content was set as the control 100, viagra increased about 10%, and silkworm moth extract 250 mg increased about 33%. It is also effective in the case of Yin Yang Kwak and Bokbunja, which are helpful for male power in other oriental medicine.

Testosterone content determination division Testosterone Content (mg / dl serum) % Increase Control 14.34 ± 1.63 100.0 Viagra 15.76 ± 1.83 109.9 Kissometer 13.20 ± 0.94 92.1 Silkworm extract sample 100mg 17.66 ± 1.13 123.2 250mg silkworm extract sample 19.04 ± 1.72 132.8 Silkworm Extract Sample 500mg 16.41 ± 1.14 114.4 Yin Yang 17.46 ± 1.38 121.8 Yin Yang Kwak + Paekuk + Bokbunja 17.16 ± 1.18 119.7

2) Comparison of Testosterone Contents of Male Moth and Male Chrysalis

Silkworms picked up several chrysalis after about two weeks of making cocoons, and only pupa was seen in the pupa but the inside was almost metamorphic, and the male moths were completely transformed and extracted from the moths. As a result, in Table 2 and in the serum, 1% of the moth extract 1% increase, 19% of the pupa extract 1% showed almost similar results.

The status of moths and chrysalis is not much different, but silkworm chrysalis are recognized as food in Korea, but silkworm moths are not classified as food. If silkworm moth extract is separated into a single substance and used as a pharmaceutical material, and the pupa is developed as a food and functional product, there is no problem.

Testosterone content in serum division Testosterone Content (mg / dl serum) % Increase Control 26.78 100.0 Male Moth Extract 1.00% 30.94 115.5 0.10% 30.40 113.5 0.01% 28.59 106.8 Can pupa extract 1.00% 31.94 119.3 0.10% 30.58 114.2 0.01% 28.86 107.8

2. Nitric oxide measurement method

25 μl of 6.5 M HCl and 25 μl of 37.5 mM sulfanilic acid were added to a test tube prepared with 100 μl of serum. After leaving for 10 minutes, 25 μl of 12.5 mM N- (1-naphthyl) -ethylenediamine was added, and then allowed to stand at room temperature for 30 minutes, followed by centrifugation at 10,000 × g for 5 minutes. Supernatants from which protein components were removed were quantified by standard calibration curves with sodium nitrate by measuring absorbance at 540 nm.

NO content in testicular tissues of rats was homogenized with 1 ml of 0.1 M phosphate buffer (pH 7.5), followed by centrifugation at 600 × g for 5 min. 100 μl of NADPII, 100 μl of 3.5 μM FAD, and 200 μl of 0.1 M phosphate buffer (pH 7.5) are added and incubated at 25 ° C. for 1 hour. Then boil for 3 minutes to stop all reactions. Griss reagent (2% sulfanylic acid / 5% H ₃ PO ₄ : 0.2% N- (1-naphthyl) -ethylenediamine / water) is then prepared in a ratio of 1: 1 and 700 μl is added. After warming again at 60 ° C. for 10 minutes, the absorbance was measured at 546 nm and quantified by a standard calibration curve with sodium nitrate.

Nitric oxide (NO) is known to play a role in blood pressure regulation, neurotransmission, blood coagulation, and immune function, and is synthesized by nitric oxide synthase (NOS) from L-arginine in various tissues and cells. NO is also known as an erectile promoter by vasodilation.

Changes in NO content were measured when various samples such as silkworm moths were administered. As shown in Table 3, it was confirmed that Yin-Yang and silkworm extract increased the NO content by about 15-20%. This proved that silkworm extract sample had some effect on vasodilation.

Nitric oxide content measurement division Testosterone Content (mg / dl serum) % Increase Control 20.21 ± 1.12 100.0 Viagra 21.28 ± 2.01 105.3 Kissometer 21.33 ± 0.42 105.5 Silkworm extract sample 100mg 21.62 ± 2.23 106.9 250mg silkworm extract sample 23.56 ± 1.35 116.6 Silkworm Extract Sample 500mg 23.42 ± 0.93 115.9 Yin Yang 23.98 ± 0.65 118.7 Yin Yang Kwak + Paekuk + Bokbunja 24.06 ± 0.22 119.0

3. Sperm Count

The testes of the rats receiving the sample are dissected and weighed. Homogenize using HBSS (hanks' balanced salt solution.) Solution and dilute by taking a certain amount from this homogenate (40 times).

Calculation = sperm count × 10 mm × dilution factor / testicle weight (g)

In male power, not only the content of male hormones and NO, but also the productivity of sperm is very important. As shown in Table 4, viagra is about 12%, silkworm extract sample 250mg shows an effect of about 41% higher than any other sample. Even if Yin-yang and Bokbunja are contained, the production of sperm is increasing.

Sperm count division Testosterone Content (mg / dl serum) % Increase Control 53.38 ± 2.18 100.0 Viagra 59.72 ± 0.80 111.9 Kissometer 51.51 ± 1.68 96.5 Silkworm male moth extract 100mg 70.22 ± 2.37 135.6 Silkworm male moth extract 250mg 75.47 ± 0.09 141.4 Silkworm male moth extract 500mg 66.47 ± 4.71 124.5 Yin Yang 70.95 ± 2.05 132.9 Yin Yang Kwak + Paekuk + Bokbunja 66.73 ± 1.11 125.0

4. Endurance Measurement

The endurance measurement was performed according to the method of Avakyan et al. (Byullen Experimental noi medisiny, 84: 375). A male ICR mouse is used as an experimental animal, and the tail of the animal is fixed upside down from the surface to a height of 10 to 12 cm on a 100 × 50 × 50 cm water bath so that the head and upper limbs of the mouse are submerged so as to induce movement. Measure the amount of time your head is submerged with maximum fatigue. At this time, the temperature of the water bath was maintained at 14 to 16 ℃, the sample was continuously administered for 4 days and the experiment was conducted only 24 hours after the final sample administration.

The silkworm powder used in this endurance experiment was a lyophilized powder of 5-day-old larvae of silkworms.

We examined the effects of endurance on masculinity or strength rather than directly related to erectile dysfunction. The animals were measured for the maximum fatigue and the time the head was submerged. As a result, as shown in Table 5, Viagra has a lower endurance than the control group, and the male moth extract of the present invention has increased the endurance more than the silkworm powder, which is about 60% more effective than the control group.

Tocopherol has also been shown to be somewhat effective in improving endurance.

Endurance measurement by forced swimming time division Control Male moth extract Pupa extract Silkworm powder Viagra Tocopherol Swimming time (minutes) 14.42 ± 3.71 23.10 ± 8.71 15.33 ± 8.71 20.37 ± 8.71 12.16 ± 8.71 17.21 ± 8.71 % Increase 100 160 106 141 84 119

As described and demonstrated in detail above, the present invention provides a method for effectively extracting erectile dysfunction and masculinity material from uncoated silkworm moth and pupa.

Silkworm moth and chrysalis of the present invention uses silkworm varieties that can be distinguished by the color of cocoons or silkworm larvae, so that silkworm chrysalis becomes a moth from the beginning. Resolved the point that could not be collected. In addition, since only a few moths can not determine whether or not mating, using a few chrysalis immediately before the moth can not only obtain a certain effect of the male moths, but also has the advantage of ensuring that the number of uncoated chrysalis.

And male moths are not allowed as food, but silkworm pupae has the advantage that it can be used as a food because it can be used as a food regardless of gender. Male moths can be used as pharmaceutical materials by purely separating substances directly related to erectile dysfunction or male history.

Claims (4)

Silkworm moth and chrysalis are characterized by separating the male and female chrysalis using silkworm varieties that can be distinguished by the color of cocoons or by the presence or absence of the pattern of silkworm larvae. A method for preparing an extract of uncoated male moth or pupa with effect. An extract of several pupa or male moths prepared by the method of claim 1. Food having a firming effect, characterized in that it comprises an extract of the pupal or male moth of claim 2. A pharmaceutical composition comprising as an active ingredient the extract of the pupal or the male moth of claim 2 in an amount effective to exhibit a sedative effect.