WO2018207810A1 - 羊膜由来原料の製造方法、化粧品の製造方法及び健康食品の製造方法 - Google Patents
羊膜由来原料の製造方法、化粧品の製造方法及び健康食品の製造方法 Download PDFInfo
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- WO2018207810A1 WO2018207810A1 PCT/JP2018/017891 JP2018017891W WO2018207810A1 WO 2018207810 A1 WO2018207810 A1 WO 2018207810A1 JP 2018017891 W JP2018017891 W JP 2018017891W WO 2018207810 A1 WO2018207810 A1 WO 2018207810A1
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- Prior art keywords
- amniotic membrane
- amnion
- placenta
- separation
- thawing
- Prior art date
Links
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Classifications
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
- A61K8/982—Reproductive organs; Embryos, Eggs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
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- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23V2200/00—Function of food ingredients
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- A—HUMAN NECESSITIES
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- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Definitions
- the present invention relates to a method for producing an amniotic material derived from amniotic membranes of mammals such as humans, pigs, horses or sheep, and a cosmetic and health food containing the amniotic material.
- Patent Document 1 discloses that a placenta of human, cow, pig or sheep is frozen and minced, thawed, washed with water, dehydrated by centrifugation or filtration, and then added with water, alkaline protease or the like. Then, the enzyme hydrolysis reaction is caused by heating and stirring. After the heating, the reaction is stopped and the reaction is stopped and left overnight at room temperature. The supernatant is collected and filtered, and the filtrate is treated with ethyl alcohol.
- Patent Document 2 discloses a placenta that has been pretreated including the removal of blood, dirt, and odorous sites, and an enzyme that contains 0.2% to 2% by weight of protease with respect to the placenta weight after the pretreatment.
- a step of vacuum packaging by interpolating into a flexible bag, and an enzyme containing protease within a range of 20 to 50 degrees at a pressure of 50 MPa to less than 200 MPa.
- Patent Document 3 discloses a method for producing a placenta extract in which human or porcine placenta is enzymatically digested with a specific heat-resistant neutral protein degrading enzyme for the purpose of eliminating the complexity of the enzymatic digestion process. It is disclosed.
- Placenta-derived materials contain ingredients such as proteins, amino acids, vitamins, minerals, enzymes, nucleic acids, and growth factors (EGF, etc.), thereby promoting skin metabolism, eliminating active oxygen, enhancing liver function, anti-inflammatory and immune It has an anti-aging effect such as activation and is used as a raw material for cosmetics and health foods.
- ingredients such as proteins, amino acids, vitamins, minerals, enzymes, nucleic acids, and growth factors (EGF, etc.
- this invention aims at providing the manufacturing method of extract etc. in which many components useful for an anti-aging effect are contained rather than the conventional placenta origin raw material, and the cosmetics and health food by which the extract etc. were mix
- the method for producing an amnion-derived material of the present invention according to claim 1 includes a pre-separation freezing step of freezing the collected placenta and amniotic membrane, and a separation in which the placenta and the amniotic membrane are thawed with running water after the pre-separation freezing step.
- the method for producing an amnion-derived material of the present invention according to claim 2 includes a pre-separation freezing step for freezing the collected placenta and amniotic membrane, and a separation for thawing the placenta and the amniotic membrane with running water after the pre-separation freezing step.
- a method for producing a cosmetic according to the present invention wherein the amniotic membrane-derived raw material obtained by the method for producing an amniotic membrane-derived raw material according to the first or second aspect is blended.
- a method for producing a health food comprising the amniotic membrane-derived material obtained by the method for producing an amnion-derived material according to the first or second aspect.
- an amnion-derived material that contains more components useful for anti-aging effects than conventional placenta-derived materials, and cosmetics and health foods containing the amniotic material-derived materials.
- the method for producing an amnion-derived material according to the first embodiment of the present invention includes a pre-separation freezing step for freezing the collected placenta and amniotic membrane, and a pre-separation freezing step before the separation for thawing the placenta and amniotic membrane with running water.
- the pre-separation thawing step draining the placenta and the amniotic membrane, the pre-separation draining step, the separation step for separating the placenta and the amniotic membrane, and the separation step, washing the amniotic membrane after the separation step Amniotic membrane washing process, Amniotic membrane draining process after the amniotic membrane flushing process, Amniotic membrane draining process after the amniotic membrane draining process, Amniotic membrane shredding process, Amniotic membrane freezing the amniotic membrane after the amniotic membrane shredding process A freezing step; an amnion thawing step for thawing the amniotic membrane after the amniotic membrane freezing step; and an extraction step for decomposing and extracting the amniotic membrane to obtain an extract after the amnion thawing step.
- placenta and amniotic membrane are those collected from horses. According to the present embodiment, it is possible to provide an amnion-derived material that contains more components useful for anti-aging effects than conventional placenta-derived materials. Moreover, according to this Embodiment, an amniotic membrane extract can be extracted efficiently.
- the method for producing an amnion-derived raw material according to the second embodiment of the present invention includes a pre-separation freezing step for freezing the collected placenta and amniotic membrane, and a pre-separation freezing step before the separation for thawing the placenta and amniotic membrane with running water.
- the pre-separation thawing step draining the placenta and the amniotic membrane, the pre-separation draining step, the separation step for separating the placenta and the amniotic membrane, and the separation step, washing the amniotic membrane after the separation step Amniotic membrane washing process, Amniotic membrane draining process after the amniotic membrane flushing process, Amniotic membrane draining process after the amniotic membrane draining process, Amniotic membrane shredding process, Amniotic membrane freezing the amniotic membrane after the amniotic membrane shredding process It has a freezing step, an amnion thawing step for thawing the amniotic membrane after the amniotic membrane freezing step, and an extraction step for decomposing and extracting the amniotic membrane to obtain an extract after the amnion thawing step.
- the cosmetic production method according to the third embodiment of the present invention blends the amniotic membrane-derived material obtained by the method for producing the amniotic membrane-derived material according to the first or second embodiment. According to this Embodiment, the cosmetics excellent in the anti-aging effect can be provided.
- the method for producing a health food according to the fourth embodiment of the present invention blends the amnion-derived material obtained by the method for producing the amnion-derived material according to the first or second embodiment. According to this Embodiment, the health food excellent in the anti-aging effect can be provided.
- FIG. 1 is a production process diagram of an amnion-derived material according to this example.
- the manufacturing method of the amniotic membrane-derived raw material (amniotic extract) according to this example is the pre-separation freezing step 10, the pre-separation thawing step 11, the pre-separation draining step 12, the separation step 13, the amnion rinsing step 14, the amnion draining step 15, Cutting step 16, amniotic membrane freezing step 17, amnion thawing step 18 and extraction step 19.
- the placenta and amniotic membrane collected immediately after parturition are immediately frozen at about -20 degrees.
- the frozen placenta and amniotic membrane are transported and stored in place. Thereby, the collected placenta and amniotic membrane can be stored in good condition.
- the placenta and amniotic membrane to be collected may be those of mammals such as humans, pigs, horses or sheep, and among them, the placenta of pigs or horses that are excellent in safety and suitable for use in cosmetics and health foods. And amniotic membrane is preferred.
- the placenta and amniotic membrane frozen in the pre-separation freezing step 10 are placed in a container, and the placenta and amniotic membrane are rapidly thawed by flowing water into the container.
- the pre-separation thawing step 11 is performed immediately before the separation step 13 is started. Also, water continues to flow after the placenta and amniotic membrane are thawed. By continuing to flow water, the water that touches the placenta and amniotic membrane is always kept clean, and the thawing water contaminated with blood or the like does not adhere to the placenta or amniotic membrane. Thereby, it can prevent that an amnion origin raw material influences the smell and color of final products, such as cosmetics or a health food.
- the placenta and amniotic membrane thawed in the pre-separation thawing step 11 are placed in a draining container such as a monkey and drained. Thereby, excess water can be removed, and workability in the subsequent process is improved.
- the amniotic membrane is separated from the placenta and amniotic membrane drained in the pre-separation draining step 12.
- the shape of the placenta and amniotic membrane varies depending on the animal species, for example, the placenta that is formed in a disk shape in the part of the uterus in the case of humans, and the scattered part formed in the whole uterus in the case of pigs and horses It is a structure called the sex placenta.
- the amniotic membrane before separation of the placenta is opened and stretched with a cutting tool such as scissors.
- placenta and amnion are separated.
- the placenta and blood vessels attached to the separated amniotic membrane are carefully removed so as not to leave behind.
- the amnion extract obtained in the extraction step 19 using the thus-separated amniotic membrane becomes an amnion-derived raw material containing more components useful for the anti-aging effect.
- final products such as cosmetics and health foods, there is little effect on the odor and color of the final product.
- the amnion separated from the placenta in the separation step 13 is washed again with water. This can more reliably prevent the placenta and blood vessels from remaining in the separated amniotic membrane.
- the amniotic membrane washed in the amniotic membrane washing step 14 is put into a draining container such as a colander to drain the water.
- a draining container such as a colander to drain the water.
- the amniotic membrane draining step 15 is followed by the amniotic membrane shredding step 16.
- the amniotic membrane cut in the amniotic shredding step 16 is drained again.
- the amniotic membrane drained in the amniotic membrane draining step 15 is cut into, for example, a 9 mm square with a shredding machine such as a mincing machine.
- the weight of the amniotic membrane shredded and drained in the amniotic shredding step 16 is measured, put in a storage container such as a plastic bag, air-tightened, sealed, and immediately put in a freezer for about ⁇ 20 Freeze the amniotic membrane at a degree. Thereby, the shredded amniotic membrane can be stored in a good state.
- the weight of the measured amniotic membrane is suppressed from variation due to moisture.
- an amnion of about 15% to 40% by weight with respect to the placenta before separation is obtained.
- the amniotic membrane frozen in the amniotic membrane freezing step 17 is thawed in the amniotic membrane thawing step 18.
- the amniotic membrane thawed in the amniotic membrane thawing step 18 is decomposed and extracted by a method such as enzyme treatment, hydrolysis treatment, acid treatment, or freeze-thawing. Thereby, an amniotic membrane extract can be obtained.
- a method such as enzyme treatment, hydrolysis treatment, acid treatment, or freeze-thawing.
- an amniotic membrane extract can be obtained.
- the amniotic membrane extract obtained in the extraction step 19 can be freeze-dried or spray-dried to obtain an amnion extract powder material to be blended in cosmetics, health foods, and the like.
- a cosmetic or health food containing the amniotic membrane extract separated from the placenta can be obtained.
- the process does not proceed to the extraction step 19, and when the thawed amniotic membrane is crushed by drying such as freeze-drying, an amnion powder raw material can be obtained.
- FIG. 2 is a diagram showing the results of quantifying index components of amniotic membrane extract produced by the process shown in FIG. 1 by enzyme immunoassay (unit: pg / g).
- the index components were cytokines such as Epidermal Growth Factor (EGF), acidic Fibroblast Growth Factor (aFGF), Hepatocyte Growth Factor (HGF), and Growth Differentiation Factor 11 (GDF-11).
- EGF Epidermal Growth Factor
- aFGF acidic Fibroblast Growth Factor
- HGF Hepatocyte Growth Factor
- GDF-11 Growth Differentiation Factor 11
- FIG. 2 (a) shows the result of quantifying the index component of the powder raw material of amniotic membrane extract.
- the raw material of the amniotic extract is extracted by mincing the amniotic membrane in minced step 16 and then passing through amniotic membrane freezing step 17 and amnion thawing step 18 and in extraction step 19 by adding purified water and neutral protease. It is manufactured by treating the temperature at 30 ° C. or higher and 70 ° C. or lower, the extraction time 1 hour or longer and 10 hours or less, and freeze-drying the obtained amniotic membrane extract.
- Example 1 uses horse equine amniotic membrane as the starting material
- Example 2 uses porcine amniotic membrane.
- FIG. 1 uses horse equine amniotic membrane as the starting material
- Example 2 uses porcine amniotic membrane.
- the result of having quantified the index component of the powder raw material of a placenta extract is also shown as a comparative example.
- the placenta extract powder raw material was produced by processing under the same conditions as in Examples 1 and 2 except that the placenta before separation was used instead of the amniotic membrane.
- the case where the placenta of the horse is used as the source material is Comparative Example 1, and the case where the placenta of the pig is used is Comparative Example 2.
- the index component is detected with a high value and a good balance as compared with Comparative Examples 1 and 2.
- GDF-11 is not detected in Comparative Examples 1 and 2 or is a low value even if detected, whereas a high value is detected in Examples 1 and 2. Since these indicator components are useful for anti-aging, by mixing the raw material of amniotic extract using amniotic membrane separated from placenta, placenta before separating amniotic membrane or placenta after separating amniotic membrane It is possible to provide cosmetics and health foods that are more excellent in anti-aging effects than when the placenta extract powder raw material used is blended. For GDF-11, the value in Example 1 is higher than that in Example 2. Therefore, when it is desired to contain a large amount of GDF-11, it is preferable to use a raw material of horse amniotic extract.
- FIG.2 (b) has shown the result of having quantified the index component of the liquid raw material of amniotic membrane extract.
- the liquid raw material of the amniotic extract is obtained by chopping the amniotic membrane into a minced shape in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and repeating the freezing and thawing a predetermined number of times in the extraction step 19. It was produced by separating the minutes.
- Example 3 is a case where horse amniotic membrane is used as a source material
- Example 4 is a case where porcine amniotic membrane is used.
- FIG.2 (b) the result of having quantified the index component of the liquid raw material of a placenta extract is also shown as a comparative example.
- the liquid material of placenta extract was produced by processing under the same conditions as in Examples 3 and 4 except that the placenta before separation was used instead of amniotic membrane.
- the case where the equine placenta is used as the source material is Comparative Example 3, and the case where the pig placenta is used is Comparative Example 4.
- the index component is detected in a well-balanced manner with a higher value than Comparative Examples 3 and 4.
- GDF-11 is not detected in Comparative Examples 3 and 4 or is a low value even if it is detected, whereas a high value is detected in Examples 3 and 4. Since these indicator components are useful for anti-aging, by adding the amniotic extract liquid material using amniotic membrane separated from placenta, placenta before separating amniotic membrane or placenta after separating amniotic membrane It is possible to provide cosmetics and health foods that are more excellent in anti-aging effect than when the liquid material of the placenta extract used is blended.
- the value in Example 3 is higher than that in Example 4. Therefore, when it is desired to contain a large amount of GDF-11, it is preferable to use a liquid raw material of horse amniotic extract.
- FIG. 3 is a diagram showing keratin water content retention data of the amniotic membrane extract produced by the process shown in FIG.
- the horizontal axis represents elapsed time (min), and the vertical axis represents keratin water content ( ⁇ s).
- “ ⁇ ” indicates that the horse's amniotic membrane is shredded into mince in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and purified water and neutral protease are added in the extraction step 19.
- This is data of an amniotic extract produced by treating the extraction temperature at 30 ° C. or higher and 70 ° C.
- Example 5 the extraction time 1 hour or longer and 10 hours or less, and performing 0.2 ⁇ m filtration.
- ⁇ and “ ⁇ ” are data of the comparative example.
- “ ⁇ ” indicates data of placenta extract obtained by treating horse placenta (Comparative Example 5), and “ ⁇ ” indicates an additive-free control (water) containing neither amniotic extract nor placenta extract. Data are shown (Comparative Example 6).
- the placenta extract of Comparative Example 5 was produced by treatment under the same conditions as Example 5 except that placenta was used instead of amniotic membrane.
- the amniotic membrane extract of Example 5 exceeded the placenta extract of Comparative Example 5 after 90 minutes from the application, and the amount of horny water exceeded that of Placenta Extract of Comparative Example 5. It can be seen that it has a longer moisture retention than the extract.
- FIG. 4 is a diagram showing the results of a usability test of the amniotic membrane extract produced by the process shown in FIG.
- the usability test is conducted for a month on 10 human monitors with lotion containing 1% amniotic membrane extract and evaluated for each item of "Moist", “Hari”, “Glossy” and “Softness”. Obtained.
- the amniotic membrane extract used in the usability test is equine amniotic membrane, and the amniotic membrane is shredded into mince in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and in the extraction step 19 It is produced by adding purified water and neutral protease, treating the extraction temperature at 30 ° C. or more and 70 ° C. or less, the extraction time at 1 hour or more and 10 hours or less, and performing 0.2 ⁇ m filtration.
- the improvement effect was confirmed in any of “moisturizing (moisturizing effect)”, “harness”, “gloss” and “softness”.
- FIG. 5 is a view showing the effect when the amniotic membrane extract produced by the process shown in FIG. 1 is applied to the back of the hand (Example 6).
- FIG. 6 is a figure which shows the effect at the time of apply
- FIG. 7 is a figure which shows the effect at the time of apply
- the amniotic membrane extract of Examples 6 and 7 was horse amniotic membrane as a starting material, and the amniotic membrane was shredded into minced in the amniotic shredding step 16, then passed through the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and in the extraction step 19 It is produced by adding purified water and neutral protease, treating the extraction temperature at 30 ° C. or more and 70 ° C. or less, the extraction time at 1 hour or more and 10 hours or less, and performing 0.2 ⁇ m filtration.
- the placenta extract of Comparative Example 7 was produced by processing under the same conditions as the amniotic membrane extract, except that the placenta was used instead of the amniotic membrane.
- FIGS. 5 and 6 show the state of the texture for each human monitor.
- FIGS. 5 (a) to (e) show the texture of human monitors A to E, respectively
- FIGS. 6 (a) to (e) show the texture of human monitors F to J, respectively.
- the left side is the textured state before application
- the right side is the textured state of the last day.
- FIG. 7A regarding the human monitor A, the left side is the textured state before application, and the right side is the textured state of the last day. Moreover, in FIG.7 (b), the upper stage shows the skin state before application
- FIG. 8 is a diagram showing the results of a drinking test of amniotic membrane extract produced by the process shown in FIG.
- the drinking test consisted of 10 human monitors for a health food (Example 8) filled with hard capsules of horse amnion extract powder material and a health food (Comparative Example 8) filled with powdered horse placenta extract powder materials. For a total of 10 days, and evaluate each item of “ease of drinking”, “smell”, “good physical condition”, “moisturizing skin”, “skin elasticity / elasticity” and “skin texture” Obtained.
- the raw material of the amniotic extract used in the drinking test was horse amniotic membrane derived from the raw material, and the amniotic membrane was shredded into mince in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, followed by the extraction step In 19, purified water and neutral protease were added, the extraction temperature was 30 ° C. to 70 ° C., the extraction time was 1 hour to 10 hours, and the resulting amniotic membrane extract was freeze-dried. is there. Moreover, the powder raw material of a placenta extract is manufactured by processing on the same conditions as Example 8 except having used the placenta instead of the amniotic membrane.
- the drinking test was performed by the following method, and the human monitor selected which of Example 8 and Comparative Example 8 was superior for each item.
- Group 1 (5 human monitors): (1) Drinking health food of Example 8 for 5 days ⁇ (2) Drinking health food of Comparative Example 8 for 5 days (until 1 month in total (1) and (2 )repeat)
- Second group (5 human monitors): (1) The health food of Comparative Example 8 was drunk for 5 days ⁇ (2) The health food of Example 8 was drunk for 5 days (until 1 month in total (1) and (2 )repeat)
- the amniotic membrane extract of Example 8 has a body sensation equivalent to that of the placenta extract of Comparative Example 8 in the items of “ease of drinking”, “odor” and “moisturizing skin”.
- the effect of improving the bodily sensation was confirmed as compared with the placenta extract of Comparative Example 8.
- FIG. 9 is a diagram showing the cell activation effect of a liquid raw material of amniotic membrane extract.
- FIG. 9 (a) shows the results for epidermal cells
- FIG. 9 (b) shows the results for dermal fibroblasts.
- the horizontal axis is the amniotic membrane extract concentration (%)
- the vertical axis is the cell activation rate (%).
- control data to which no amniotic extract is added is shown as a comparative example at the left end.
- the liquid raw material of the amniotic extract is obtained by chopping the amniotic membrane into a minced shape in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and repeating the freezing and thawing a predetermined number of times in the extraction step 19. It was produced by separating the minutes. Horse amniotic membrane is used as a raw material.
- amniotic membrane extract obtained by the production method according to this example has a cell activation effect on epidermal cells and dermal fibroblasts.
- FIG. 10 is a diagram showing an action of promoting collagen production from human fibroblasts by a liquid raw material of amniotic membrane extract.
- the horizontal axis represents the amniotic membrane extract concentration (%), and the vertical axis represents the collagen production rate (%).
- the liquid raw material of the amniotic extract is obtained by chopping the amniotic membrane into a minced shape in the amniotic shredding step 16, followed by the amniotic membrane freezing step 17 and the amniotic membrane thawing step 18, and repeating the freezing and thawing a predetermined number of times in the extraction step 19. It was produced by separating the minutes. Horse amniotic membrane is used as a raw material.
- amniotic membrane extract obtained by the production method according to the present example has an action of promoting collagen production from fibroblasts.
- an amnion-derived material of the present invention it is possible to provide an amnion-derived material that contains more components useful for the anti-aging effect than conventional placenta-derived materials.
- cosmetics, health foods and the like in which this amniotic membrane-derived material is blended can be provided.
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Abstract
Description
例えば特許文献1には、ヒト、ウシ、ブタ又はヒツジの胎盤を凍結してミンチ化し、融解し、水洗した後、遠心または濾過により脱水して得た脱水胎盤を、水、アルカリプロテアーゼ等を加えて、加熱、撹拌を行うことで酵素加水分解反応を起こし、さらに、加熱した後、反応を停止させて室温で一晩放置し、上澄液を採取して濾過し、濾液をエチルアルコールで処理してエチルアルコール溶出液にし、溶出液を減圧濃縮したものを凍結乾燥して粉末にする胎盤エキスからなる抗アレルギー剤の製造方法が開示されている。
また、特許文献2には、血液、汚物及び臭い部位の除去を含む前処理をした胎盤と、前処理後の胎盤重量に対して0.2重量%~2重量%のプロテアーゼを含む酵素とを、可撓性を有する袋体に内挿して真空包装する工程と、真空包装した袋体を、50MPa以上~200MPa未満の圧力で、20度~50度の範囲内でプロテアーゼを含む酵素の至適温度に制御した状態で、1日~3日保持する抽出工程と、袋体内の内容物からプラセンタエキス原液を分離する固液分離工程とを有するプラセンタエキスの製造方法が開示されている。
また、特許文献3には、酵素消化過程における煩雑さの解消を目的として、ヒト又はブタの胎盤を、特定の耐熱性中性タン白質分解酵素を使用して酵素消化するプラセンタエキスの製造方法が開示されている。
しかし、さらにアンチエイジング効果に優れた化粧品や健康食品等を提供するため、有用成分がより多く含まれるエキス等の開発が期待されている。
請求項2記載の本発明の羊膜由来原料の製造方法は、採集した胎盤及び羊膜を凍結する分離前凍結工程と、前記分離前凍結工程の後に、前記胎盤及び前記羊膜を流水にて解凍する分離前解凍工程と、前記分離前解凍工程の後に、前記胎盤及び前記羊膜の水切りを行う分離前水切工程と、前記分離前水切工程の後に、前記胎盤と前記羊膜を分離する分離工程と、前記分離工程の後に、前記羊膜を水洗いする羊膜水洗工程と、前記羊膜水洗工程の後に、前記羊膜の水切りを行う羊膜水切工程と、前記羊膜水切工程の後に、前記羊膜を細断する羊膜細断工程と、前記羊膜細断工程の後に、前記羊膜を凍結する羊膜凍結工程と、前記羊膜凍結工程の後に、前記羊膜を解凍する羊膜解凍工程と、前記羊膜解凍工程の後に、前記羊膜を分解抽出してエキスを得る抽出工程とを有し、前記抽出工程は、前記羊膜の凍結と融解を所定回数繰り返して水溶性画分を分離するものであり、前記胎盤及び前記羊膜は、ウマから採集されたものであることを特徴とする。
請求項3記載の本発明の化粧品の製造方法は、請求項1又は請求項2に記載の羊膜由来原料の製造方法によって得られた羊膜由来原料を配合することを特徴とする。
請求項4記載の本発明の健康食品の製造方法は、請求項1又は請求項2に記載の羊膜由来原料の製造方法によって得られた羊膜由来原料を配合することを特徴とする。
本実施の形態によれば、従来の胎盤由来原料よりもアンチエイジング効果に有用な成分が多く含まれる羊膜由来原料を提供できる。また、本実施の形態によれば、効率よく羊膜エキスを抽出できる。
本実施の形態によれば、従来の胎盤由来原料よりもアンチエイジング効果に有用な成分が多く含まれる羊膜由来原料を提供できる。また、本実施の形態によれば、効率よく羊膜エキスを抽出できる。
本実施の形態によれば、アンチエイジング効果に優れた化粧品を提供できる。
本実施の形態によれば、アンチエイジング効果に優れた健康食品を提供できる。
本実施例による羊膜由来原料(羊膜エキス)の製造方法は、分離前凍結工程10、分離前解凍工程11、分離前水切工程12、分離工程13、羊膜水洗工程14、羊膜水切工程15、羊膜細断工程16、羊膜凍結工程17、羊膜解凍工程18及び抽出工程19を有する。
なお、採集する胎盤及び羊膜は、ヒト、ブタ、ウマ又はヒツジなどの哺乳類動物のものであればよいが、その中でも安全性に優れ、かつ化粧品及び健康食品の用途に適したブタ又はウマの胎盤及び羊膜とすることが好ましい。
また、胎盤及び羊膜が解凍された後も水を流し続ける。水を流し続けることにより、胎盤や羊膜に触れる水が常に清浄な状態に保たれ、血液等で汚染された解凍用水が胎盤や羊膜に付着することがない。これにより、羊膜由来原料が化粧品又は健康食品といった最終製品のにおいや色に影響することを防止できる。
胎盤及び羊膜の形状は動物種によって異なり、例えば、ヒトの場合は子宮の一部に円盤状に形成される盤状胎盤、ブタ及びウマの場合は子宮内の全体に散在して形成される散在性胎盤と呼ばれる構造である。胎盤から羊膜を分離する際は、ヒトの場合は胎盤が一つの大きな塊となっているので作業が明確であるが、散在性胎盤であるブタ及びウマの場合はそうではないため、以下の手順で行う。
まず、胎盤を分離する前の羊膜をハサミなどの切断具で切り開いて伸ばす。次に、胎盤と羊膜を分離する。次に、分離した羊膜に付着した胎盤及び血管を取り残しがないように注意して除去する。これにより、胎盤と羊膜を効率よく分離することができ、胎盤や血管が付着していない羊膜のみを抽出に供することができる。このように分離された羊膜を用いて抽出工程19で得られた羊膜エキスは、アンチエイジング効果に有用な成分がより多く含まれた羊膜由来原料となる。また、化粧品や健康食品といった最終製品に配合する場合に、最終製品のにおいや色への影響が少ない。
羊膜水切工程15の後に羊膜細断工程16を行う。羊膜細断工程16で細断した羊膜は、再度水切りする。再度水切りを行うことで水分管理の精度を高め、羊膜の重量の水分によるばらつきを更に抑制し、羊膜由来原料の品質をより一層向上させることができる。
なお、例えばウマの胎盤からは、分離前の胎盤に対する重量%で約15%~40%重量の羊膜が得られる。
羊膜凍結工程17で凍結した羊膜は、羊膜解凍工程18において解凍する。
抽出工程19で得られた羊膜エキスを遠心分離又はろ過することにより、化粧品や健康食品等に配合する羊膜エキスの液体原料とすることができる。
また、抽出工程19で得られた羊膜エキスを凍結乾燥又は噴霧乾燥することにより、化粧品や健康食品等に配合する羊膜エキスの粉末原料とすることができる。
羊膜エキスの液体原料又は粉末原料を所定量配合することにより、胎盤から分離した羊膜のエキスが配合された化粧品又は健康食品を得ることができる。
なお、羊膜解凍工程18の後、抽出工程19には移行せず、解凍した羊膜についてフリーズドライ等の乾燥を行って破砕した場合には、羊膜の粉末原料を得ることができる。
指標成分は、サイトカイン類であるEpidermal Growth Factor(EGF)、acidic Fibroblast Growth Factor(aFGF)、Hepatocyte Growth Factor(HGF)、Growth Differentiation Factor 11(GDF-11)とした。
また、図2(a)には、比較例として、胎盤エキスの粉末原料の指標成分を定量した結果を併せて示している。胎盤エキスの粉末原料は、羊膜の代わりに分離前の胎盤を用いた以外は実施例1、2と同条件で処理することにより製造したものである。由来原料にウマの胎盤を用いた場合を比較例1、ブタの胎盤を用いた場合を比較例2としている。
また、GDF-11については、実施例2よりも実施例1のほうが高い値となっている。したがって、GDF-11を多く含ませたい場合は、ウマの羊膜エキスの粉末原料を用いることが好ましい。
また、図2(b)には、比較例として、胎盤エキスの液体原料の指標成分を定量した結果を併せて示している。胎盤エキスの液体原料は、羊膜の代わりに分離前の胎盤を用いた以外は実施例3、4と同条件で処理することにより製造したものである。由来原料にウマの胎盤を用いた場合を比較例3、ブタの胎盤を用いた場合を比較例4としている。
また、GDF-11については、実施例4よりも実施例3のほうが高い値となっている。したがって、GDF-11を多く含ませたい場合は、ウマの羊膜エキスの液体原料を用いることが好ましい。
図中「▲」は、羊膜細断工程16においてウマの羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において精製水と中性プロテアーゼを加えて、抽出温度を30度以上70度以下、抽出時間を1時間以上10時間以下として処理し、0.2μmのろ過を行うことにより製造した羊膜エキスのデータである(実施例5)。
また、図中「■」及び「◆」は、比較例のデータである。「■」は、ウマの胎盤を処理して得られた胎盤エキスのデータを示し(比較例5)、「◆」は、羊膜エキスも胎盤エキスも含まれていない無添加のコントロール(水)のデータを示している(比較例6)。なお、比較例5の胎盤エキスは、羊膜の代わりに胎盤を用いた以外は実施例5と同条件で処理することにより製造したものである。
使用感テストは、羊膜エキスが1%配合された化粧水について、ヒトモニター10名を対象として一ヶ月行い、「しっとり」、「ハリ」、「つや」及び「やわらかさ」の各項目について評価を得た。
使用感テストに使用した羊膜エキスは、由来原料をウマの羊膜とし、羊膜細断工程16において羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において精製水と中性プロテアーゼを加えて、抽出温度を30度以上70度以下、抽出時間を1時間以上10時間以下として処理し、0.2μmのろ過を行うことにより製造したものである。
実施例6、7の羊膜エキスは、由来原料をウマの羊膜とし、羊膜細断工程16において羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において精製水と中性プロテアーゼを加えて、抽出温度を30度以上70度以下、抽出時間を1時間以上10時間以下として処理し、0.2μmのろ過を行うことにより製造したものである。
また、比較例7の胎盤エキスは、羊膜の代わりに胎盤を用いた以外は羊膜エキスと同条件で処理することにより製造したものである。
効果は、羊膜エキスを塗布する群としてヒトモニターA~Eの5名、胎盤エキスを塗布する群としてヒトモニターF~Jの5名を対象として、以下の手順にて5日間行うことで確認した。
1.塗布前に肌の肌理を撮影する。
2.その後、1日1回手の甲に羊膜エキス又は胎盤エキスを塗布する。なお、ヒトモニターAについては、足にも羊膜エキスを塗布する。
3.最終日の同時間に再度肌の肌理を撮影する。
図5及び図6に示すように、羊膜エキスを手の甲に塗布した結果、胎盤エキスを手の甲に塗布した結果と同等以上に、肌のきめ細かさが向上することが確認された。
図7(a)に示すように、羊膜エキスを足に塗布した結果、肌の乾燥が改善し肌のきめ細かさが向上することが確認された。
また、図7(b)に示すように、羊膜エキスを足に塗布した結果、湿疹が改善することが確認された。
飲用テストは、ウマの羊膜エキスの粉末原料をハードカプセルに充填した健康食品(実施例8)と、ウマの胎盤エキスの粉末原料をハードカプセルに充填した健康食品(比較例8)について、ヒトモニター10名を対象として計10日間行い、「飲みやすさ」、「におい」、「体調の良い変化」、「肌の潤い」、「肌のハリ・弾力」及び「肌のキメ」の各項目について評価を得た。
飲用テストに使用した羊膜エキスの粉末原料は、由来原料をウマの羊膜とし、羊膜細断工程16において羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において精製水と中性プロテアーゼを加えて、抽出温度を30度以上70度以下、抽出時間を1時間以上10時間以下として処理し、得られた羊膜エキスを凍結乾燥することにより製造したものである。また、胎盤エキスの粉末原料は、羊膜の代わりに胎盤を用いた以外は実施例8と同条件で処理することにより製造したものである。
飲用テストは、以下の方法で行い、各項目について実施例8と比較例8のどちらが優れているかをヒトモニターに選択してもらった。
第1グループ(ヒトモニター5名):(1)実施例8の健康食品を5日間飲用→(2)比較例8の健康食品を5日間飲用(計1ヶ月となるまで(1)と(2)を繰り返す)
第2グループ(ヒトモニター5名):(1)比較例8の健康食品を5日間飲用→(2)実施例8の健康食品を5日間飲用(計1ヶ月となるまで(1)と(2)を繰り返す)
羊膜エキスの液体原料は、羊膜細断工程16において羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において凍結と融解を所定回数繰り返して水溶性画分を分離することにより製造したものである。由来原料にはウマの羊膜を用いている。
羊膜エキスの液体原料は、羊膜細断工程16において羊膜をミンチ状に細断した後、羊膜凍結工程17及び羊膜解凍工程18を経て、抽出工程19において凍結と融解を所定回数繰り返して水溶性画分を分離することにより製造したものである。由来原料にはウマの羊膜を用いている。
11 分離前解凍工程
12 分離前水切工程
13 分離工程
14 羊膜水洗工程
15 羊膜水切工程
16 羊膜細断工程
17 羊膜凍結工程
18 羊膜解凍工程
19 抽出工程
Claims (4)
- 採集した胎盤及び羊膜を凍結する分離前凍結工程と、
前記分離前凍結工程の後に、前記胎盤及び前記羊膜を流水にて解凍する分離前解凍工程と、
前記分離前解凍工程の後に、前記胎盤及び前記羊膜の水切りを行う分離前水切工程と、
前記分離前水切工程の後に、前記胎盤と前記羊膜を分離する分離工程と、
前記分離工程の後に、前記羊膜を水洗いする羊膜水洗工程と、
前記羊膜水洗工程の後に、前記羊膜の水切りを行う羊膜水切工程と、
前記羊膜水切工程の後に、前記羊膜を細断する羊膜細断工程と、
前記羊膜細断工程の後に、前記羊膜を凍結する羊膜凍結工程と、
前記羊膜凍結工程の後に、前記羊膜を解凍する羊膜解凍工程と、
前記羊膜解凍工程の後に、前記羊膜を分解抽出してエキスを得る抽出工程とを有し、
前記抽出工程は、前記羊膜に精製水及び中性プロテアーゼを加え、抽出温度を30度以上70度以下、抽出時間を1時間以上10時間以下としたものであり、
前記胎盤及び前記羊膜は、ウマから採集されたものであることを特徴とする羊膜由来原料の製造方法。 - 採集した胎盤及び羊膜を凍結する分離前凍結工程と、
前記分離前凍結工程の後に、前記胎盤及び前記羊膜を流水にて解凍する分離前解凍工程と、
前記分離前解凍工程の後に、前記胎盤及び前記羊膜の水切りを行う分離前水切工程と、
前記分離前水切工程の後に、前記胎盤と前記羊膜を分離する分離工程と、
前記分離工程の後に、前記羊膜を水洗いする羊膜水洗工程と、
前記羊膜水洗工程の後に、前記羊膜の水切りを行う羊膜水切工程と、
前記羊膜水切工程の後に、前記羊膜を細断する羊膜細断工程と、
前記羊膜細断工程の後に、前記羊膜を凍結する羊膜凍結工程と、
前記羊膜凍結工程の後に、前記羊膜を解凍する羊膜解凍工程と、
前記羊膜解凍工程の後に、前記羊膜を分解抽出してエキスを得る抽出工程とを有し、
前記抽出工程は、前記羊膜の凍結と融解を所定回数繰り返して水溶性画分を分離するものであり、
前記胎盤及び前記羊膜は、ウマから採集されたものであることを特徴とする羊膜由来原料の製造方法。 - 請求項1又は請求項2に記載の羊膜由来原料の製造方法によって得られた羊膜由来原料を配合することを特徴とする化粧品の製造方法。
- 請求項1又は請求項2に記載の羊膜由来原料の製造方法によって得られた羊膜由来原料を配合することを特徴とする健康食品の製造方法。
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