WO2009002982A2 - Fluide biologique de gastéropode, procédé de fabrication et de raffinage, et utilisation - Google Patents

Fluide biologique de gastéropode, procédé de fabrication et de raffinage, et utilisation Download PDF

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Publication number
WO2009002982A2
WO2009002982A2 PCT/US2008/068047 US2008068047W WO2009002982A2 WO 2009002982 A2 WO2009002982 A2 WO 2009002982A2 US 2008068047 W US2008068047 W US 2008068047W WO 2009002982 A2 WO2009002982 A2 WO 2009002982A2
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WO
WIPO (PCT)
Prior art keywords
gastropod
biological fluid
cosmeceutical
skin care
care composition
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PCT/US2008/068047
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English (en)
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WO2009002982A3 (fr
Inventor
William Wang
Jun Yi
Sheng Ke
Maria Halmela
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Pharmacom International Inc.
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Application filed by Pharmacom International Inc. filed Critical Pharmacom International Inc.
Priority to CN200880104094A priority Critical patent/CN101795697A/zh
Publication of WO2009002982A2 publication Critical patent/WO2009002982A2/fr
Publication of WO2009002982A3 publication Critical patent/WO2009002982A3/fr
Priority to US12/643,751 priority patent/US20100233111A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1825—Fibroblast growth factor [FGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
    • A—HUMAN NECESSITIES
    • 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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60—Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • 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
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • 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
    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
    • A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
    • A—HUMAN NECESSITIES
    • 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
    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
    • A61K8/925—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of animal origin
    • A—HUMAN NECESSITIES
    • 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
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin

Definitions

  • Te present technology generally relates to skin care compositions, cosmeceutical compositions and methods of making or using the same. More specifically, the presently described technology generally relates to skin care compositions, cosmocuetical compositions as well as methods of making and using the same utilizing a unique composition recovered and refine from a gastropod, namely Helix Aspersa M ⁇ ller, among others.
  • the present technology provides a unique method of collecting and refining a gastropod biological fluid from a live gastropod.
  • the method includes, for example, the steps of stimulating the gastropod to increase the secretion of the desired biological fluid.
  • Methods of stimulating the gastropod include, for example, exposing the gastropod to sound vibration; exposing the gastropod to low hyperbaric pressure; exposing the gastropod to thermal puncture; exposing the gastropod to a physical stimulus; exposing the gastropod to ionizing radiation; exposing the gastropod to gravity; or inverting the gastropod.
  • the method further includes the step of collecting the secreted fluid and centrifuging the secreted fluid to create a supernatant.
  • the present technology provides one or more cosmeceuticals, including an effective amount of the collected and refined gastropod biological fluid.
  • the recovered and refined gastropod biological fluid e.g. the filtrate or supernatant
  • the gastropod biological fluid may include about 10% to about 20% of the one or more skin care compositions, formulations or cosmeceutical described herein.
  • the compositions may be used for skin repair and rejuvenation, treatment of wounds, acne, and scars, and/or may be used as a sunscreen, an anti- aging cream, an anti-wrinkle cream, a moisturizing cream, or as a whitening cream.
  • the presently described compositions may further include an effective amount of squalane and an effective amount of hyaluronic acid, among other additives.
  • a cosmeceutical, composition or formulation that promotes skin rejuvenation and healing comprising at least about 15% by weight of a gastropod biological filtrate.
  • the present technology provides a gastropod biological fluid including about 0.002% to about 0.1% uronic acid; about 0.003% to about 0.5% hexozamine; about 0.001 to about 0.1% acetyl; about 0.001 to about 0.1% sulfate; about 0.02% to about 1 .0% of at least one mucopolysaccharide; about 0.05% to about 0.5% of at least one fibroblast growth factor; about 0.01 % to about 0.1 % of at least one enzyme; about 0.01 % to about 1 .0% hyaluronic acid; about 0.001 % to about 0.2% of at least one oxygen carrier; about 0.005% to about 0.1 % of at least one high molecular weight protein; about 0.0001 to about 0.01 % of at least one trace element; about 0.005% to about 0.1% of at least one high molecular weight antioxidant compound; about 0.005% to about 0.1 % of at least one low molecular weight compound; and/or about 80% to about
  • the present technology provides a composition, cosmeceutical or formulation having at least about 1 % to at least about 30% of a gastropod biological fluid.
  • the gastropod biological fluid may be a supernatant or a filtrate containing enriched amounts of glycosaminoglycans.
  • the present technology provides a composition, cosmeceutical, or formulation having about 10.0% to about to about 25.0% of a gastropod excreted fluid.
  • the composition, cosmeceutical, or formulation further include about 0.5% to about 10.0% stearic acid; about 0.1 % to about 1 .0% single stearic acid glyceide; about 1 .0% to about 10.0% of at least one oil; about 0.1 % to about 1 .0% octadecanol; about 1 .0% to about 10.0% glycerin; about 0.02% to about 2.0% potassium hydroxide; and/or about 0.2% to about 2.0% of a preservative.
  • Fig. 1 is a picture of an SDS page gel showing the digestion of the gastropod biological fluid GAG with heparin lyase I, heparin lyase II, or heparin lyase III in comparison to a control.
  • Fig.2 is a set of pictures depicting histological staining of the mantle of
  • Fig. 1 -2 depicts WPA staining; Fig. 2-2 depicts nNOS staining; Fig. 2-3 depicts staining of proteoglycans with carboxyl groups; Fig. 2-4 depicts carboxylic acid proteoglycan staining; Fig. 2-5 depicts other glycoproteins staining; Fig. 2-6 depicts -D-glucose and ⁇ -D-mannose staining; Fig. 2-7 depicts oligosaccharides staining; Fig. 2-8 depicts ⁇ -D-gal and (1 -3)-D-Gal-Nac staining; Fig. 2-9 depicts ⁇ -L- fucose staining; and Fig. 2-10 depicts N-acetyl-D-galactosamine or D-galactose staining.
  • Fig. 4 illustrates histological staining of fibronectin assembly in CHO-K1 cells treated with gastropod biological fluid (SJ) of the present technology.
  • FIG. 5 illustrate histological staining of human dermal fibroblast morphology after treatment with gastropod biological fluid of the present invention.
  • Fig. 6 is a line graph depicting the antioxidant qualities of gastropod biological fluid (Snail serum).
  • Fig. 7 is a line graph depicting the moisturizing qualities of formulations of a moisturizing liquid containing gastropod biological fluid.
  • Fig. 8 is a graph depicting the long term (22 day) moisturizing effect of formulation containing gastropod biological fluid on healthy test subject's skin.
  • Fig. 9 are three dimensional graphs showing the effects of anti-wrinkle cream on wrinkle depths for groups treated with a control cream (A, B, C) and an anti- wrinkle cream containing gastropod biological fluid of the present technology (D, E, F). Measurements were taken at 0 days treatment (A, D), 15 days treatment (B, E) or 30 days treatment (C, F).
  • the present technology relates generally to a unique method collecting and refining a gastropod biological fluid from live gastropods and the use of such gastropod biological fluid in compositions, cosmeceuticals, or formulations that provide, for example, beneficial skin care treatments and therapeutic outcomes.
  • the naturally-based compositions, cosmeceuticals, and/or formulations may trigger the regeneration of damaged cells; prevent or reduce scarring, keratosis, psoriasis scales, as well as all types of skin blemishes; prevent or reduce the appearance of acne; prevent or reduce the signs of wrinkles; and can replenish and/or moisturize the lipid barrier of the skin.
  • compositions, cosmeceuticals, and/or formulations are based on natural ingredients containing preferably little or no chemicals, detergents or fillers. In doing so, it is believed (although not wanting to be bound by any particular theory) that the unique compositions having such natural ingredients provide antimicrobial, protective, moisturizing, repairing, and/or renewing properties to the skin, including components or component systems that mimic the intracellular structure of skin and are essential for normal skin function.
  • the present technology provides a method of collecting and refining a gastropod biological fluid from a live gastropod.
  • At least one advantage of the present technology is the ability to use the gastropod repeatedly for collection of the biological fluid since the process preferably does not harm the gastropod.
  • Gastropods secrete the biological fluid to be collected and refined when under stress in order to protect, moisturize, repair and renew its own skin membrane.
  • the method comprises the steps of stimulating a live gastropod to increase biological fluid secretion; collecting the secreted fluid; centrifuging the secreted fluid to create a supernatant; and filtering the supernatant to recover a filtrate. This filtrate contains an enhanced amount of one or more unique gastropod glycosaminoglycans.
  • a gastropod is a member of the Phylum Mollusca class
  • Gastropoda including both Helix (shelled) and non-Helix (shell-less) members whose glandular secreted fluid is used as the active ingredient in the practice of the present technology.
  • Suitable gastropods include, for example, Helix pomatia, Helix hortensis, Helix nemoralis, Helix cardidula, Helix tchthyomma (also referred to as Helix campylea), Helix fructicicola (also referred to as Helix se uca), Helix strigella, Helix fruticum, Helix bidens, Helix arbostorum, Helix rotundata, Helix aculeata, Helix pulchella, Helix personata, Helix holoserica, Helix aperta, Helix parnassia, Helix alonensis, Helix candidissima, Helix pisana, and Helix gualteviana.
  • gastropoda A more extensive discussion of gastropoda may be found in Invertebrate Zoology (5th Ed.), 864 p. Robert D. Barnes, (Saunders College Publishing, Inc., Troy, Mich., 1987).
  • a particularly suitable gastropod is Helix Aspersa M ⁇ ller.
  • another suitable gastropod is Helix pomatia.
  • At least one method of the presently described technology comprises stimulating a live gastropod to increase biological fluid secretion.
  • Methods to stimulate a live gastropod may include any methods that do not harm the live gastropod while increasing the biological fluid output of the gastropod, allowing the gastropod to be reused for biological fluid collection. Stimulation causes the gastropod to increase biological fluid secretions naturally produced by the mucinous, albuminous, and salivary glands.
  • These methods include, but are not limited to, exposing the gastropod to sound vibrations, exposing the gastropod to low hyperbaric pressure, exposing the gastropod to thermal punctures, exposing the gastropod to hypoxic conditions, exposing the gastropod to gravitation force, inverting the gastropod to alter its orientation, and exposing the gastropod to a physical stimulus.
  • Exposure of a gastropod to sound vibrations includes methods known in art to expose the gastropod to frequencies in the range of about 5kHz, to about 25kHz, alternatively about 10KHz to about 2OkHz, alternatively between about 15kHz to about 2OkHz for about 1 second to about 60 seconds, alternatively between about 5 seconds to about 30 seconds, more suitably about 10 seconds.
  • the gastropod may be exposed to the sound frequencies of about 5kHz, about 10kHz, about 12kHz, about 15kHz, about 2OkHz, for a period of at least about 5 seconds, at least about 10 seconds, at least about 15 seconds, at least about 20 seconds to increase secretion of the biological fluid.
  • a particularly suitable method of exposing a gastropod to sound vibrations includes exposing the gastropod to ultrasonic irradiation by an ultrasonic generator UP 200 H horn type (2OkHz with maximum wave amplitude of 210 ⁇ m and a maximum nominal power output of 460W) equipped with a radial Sonotrode S3 (3mm diameter) for about 10 seconds.
  • the gastropod may be stimulated to increase production of the biological fluid to be collected and refined by exposing the gastropod to low hyperbaric pressure.
  • Any suitable means known in the art may be used including hyperbaric oxygen therapy (HBOT) chambers.
  • HBOT hyperbaric oxygen therapy
  • Suitable HBOT chambers may expose the gastropod to pressures of about 3 pounds per square inch (psi) to about 8 psi, more suitably about 4 psi (1 .27 ATA (atmospheres absolute) 8.92 FSW) to about 7.35 psi (1 .5 ATA 16.38 FSW) with an oxygen concentration of at least about 90%, more suitably at least about 95%, more suitably at least about 100%.
  • the gastropod is exposed to the low hyperbaric pressure for at least about 5 seconds, more suitable at least about 10 seconds, more suitably at least about 12 seconds, more suitably at least about 15 seconds to stimulation the gastropod to produce the biological fluid.
  • Stimulating a gastropod to increase biological fluid secretion may also include inverting a gastropod for a specified amount of time.
  • a gastropod may be inverted by hanging the gastropod by its tail with its head toward the ground for at least about 5 seconds, at least about 7 seconds, at least about 10 seconds, at least about 30 seconds, at least about 1 minute, at least about 1 .5 minutes, at least about 2 minutes.
  • minipliers may be used to clip to the tail of the gastropod and invert the snail in the air for about 10 seconds to about 2 minutes.
  • a straw may be used to collect the biological fluid along the ventral surface of the gastropod.
  • stimulating a gastropod to increase biological fluid secretion may include exposing the gastropod to gravity.
  • the gravitational force causes an increase in biological fluid secretion in the sinus region of the gastropod, such as the buccal or cardiac sinus.
  • the biological fluid may be extracted using a syringe to remove the biological liquid.
  • gastropods may also be exposed to ionizing radiation, either x-rays or gamma rays, for about 0.5 seconds, for about 1 .0 seconds, for about 2.0 seconds. Gastropods may also be exposed to hypoxic conditions to stimulate fluid secretion. The gastropod is exposed to oxygen depletion or environmental hypoxia for at least about 10 seconds, at least about 12 seconds, at least about 15 seconds to increase production of secretion. Any method of oxygen depletion or environmental hypoxia known by one familiar in the art may be used.
  • stimulating a gastropod to increase biological fluid secretion can be done, for example, by exposing the gastropod to physical stimulation.
  • a suitable means of physically stimulating a gastropod is centrifuging the gastropod. Centrifuging is carried out under conditions which will not break the shell of the shelled gastropod or if a shell-less gastropod, under conditions in which the gastropod remains alive and is not crushed.
  • the gastropod is centrifuged for a period of time at a temperature and gravity (G) sufficient to cause the gastropod to secrete fluids.
  • G temperature and gravity
  • the force required to stimulate fluid production in a gastropod is suitable less than about 7G, more suitable less than about 6 G, more suitably between about 1 G and about 6G, more suitably between about 2G and about 5 G for about 2 minutes to about 10 minutes.
  • One suitable embodiment includes, for example, centrifuging the gastropod at or about 2G for about 10 minutes.
  • the gastropod is centrifuged at temperatures of about 10 ° C or greater, more suitably 15 ° C or greater, more suitably 20 ° C or greater, more suitably between about 10 ° C and about 35 ° C, more suitably between about 20 ° C and about 35 ° C.
  • An alternative method of centrifugation includes pulsating the gastropod for about 3 to about 4 pulsations during the centrifugation.
  • a pulsation includes accelerating the centrifuge to the desired G force, decelerating the centrifuge and accelerating the centrifuge again.
  • a suitable embodiment of this method includes centrifuging the gastropod at about 2 G for a period of about 10 minutes with about 3 or about 4 pulsations being performed during this time period.
  • the method of collecting a gastropod biological fluid after stimulating the gastropod may be by any means known to one familiar in the art.
  • a straw may be used to siphon the biological fluid from the ventral surface of the gastropod.
  • the collected biological gastropod secreted fluid may be centrifuged at about 200 rpm to about 5000 rpm, more suitably about 2000 rpm in a for at least about 2 minutes, alternatively at least about 5 minutes, alternatively at least about 2 minutes to about 10 minutes to remove any large particles and to produce a supernatant.
  • the supernatant is decanted and filtered through a microporous filter having a pore size of about 0.1 to about 1 micron to produce a filtrate. Filtration may be carried out under pressure to speed up the filtration process.
  • the filtrate containing the active ingredients in the gastropod biological fluid is then recovered.
  • the method of collecting a biological fluid from a live gastropod further includes fasting the gastropod for at least about 1 day to about 5 days, alternatively about 1 day to about 2 days before stimulation of the gastropod.
  • fasting of the gastropod decreases the amount of toxins that may be present in the secreted fluids produced as a byproduct of the food the gastropod eats.
  • the toxins may be removed from the secreted fluid by methods known by those familiar in the art to inactivate the toxins, for example
  • the present technology further provides the gastropod biological fluid including a supernatant and a filtrate made by the process described above.
  • the gastropod biological fluid contains a mixture of components that among other properties provide beneficial healing and rejuvenating effects to human skin.
  • the gastropod biological fluid may, for example, promote the immune system to clear our dead and dying cells, trigger the production of new cells, rebuilds the skin matrix by repairing, moisturizing, and protecting dermal elements.
  • the gastropod biological fluid includes about 0.001 % to about 0.1 % uronic acid; about 0.003% to about 0.3% hexozamine; about 0.001 to about 0.1 % acetyl; about 0.001 % to about 0.1% sulfate; about 0.02% to about 5.0% of at least one mucopolysaccharide; about 0.05% to about 0.5% of at least one fibroblast growth factor; about 0.01 % to about 0.1% of at least one enzyme; about 0.01 % to about 1 .0% hyaluronic acid; about 0.001 % to about 0.1 % of at least one oxygen carrier; about 0.001% to about 0.1% of at least one high molecular weight protein; about 0.0001 to about 0.01% of at least one trace element; about 0.005% to about 0.1 % of at least one high molecular weight antioxidant compound; about 0.005% to about 0.1 % of at least one low molecular weight antioxidant compound; and/or about 80% to about 98% water.
  • the at least one mucopolysaccharide may include glycosaminoglycans
  • the mucopolysaccharide is a novel glycosaminoglycan produced from Helix Aspersa M ⁇ ller having a molecular weight of about 29,000 Da, calculated based on viscometery, and a uniform repeating disaccharide structure of structure of (1— 14)-2- acetyl, 2-deoxy- -D-glucopyranose (1 — *4)-2-sulfo- ⁇ -L-idopyranosyluronic acid.
  • This polysaccharide represents a unique and believed novel glycosaminoglycan. It is related to the heparin and heparan sulfate families of glycosaminoglycans, but is distinctly different from all known members of these classes of glycosaminoglycans.
  • the structure of this polysaccharide has an adjacent ⁇ /-acetylglucosamine and 2-sulfo- iduronic acid residues.
  • the combination of a core protein and a specific glycosaminoglycan generates a unique proteoglycan with a precise developmental patter.
  • This glycosaminoglycan represents approximately about 3% to about 5% of the dry weight of the snail's soft body tissues, suggesting important biological roles for the survival of this organism.
  • the gastropod glycosaminoglycan tightly binds divalent cations, such as copper (II).
  • the mucopolysaccharide is at least about 0.02%, at least about 0.2%, at least about 0.3%, at least about 0.5%, at least about 0.8, at least about 1 %, at least about 2%, at least about 3%, or at least about 4% of the composition of the gastropod biological fluid.
  • the fibroblast growth factors are a family of structurally related polypeptide growth factors, currently consisting of 23 members having a heparin- binding domain. FGFs are known to play a role in angiogenesis, wound healing, and embryonic development.
  • the at least one fibroblast growth factor (FGF) of the present technology may be chosen from a group comprising, for example, FGF1 , FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGF10, FGF1 1 , FGF12, FGF13, FGF14, FGF15, FGF16, FGF17, FGF18, FGF19, FGF20, FGF21 , FGF22, FGF23 or a combination thereof.
  • at least one fibroblast growth factor utilized in the practice of the present technology is FGF1 , FGF4, FGF7, FGF10 or a combination thereof.
  • the unique gastropod biological fluid of the present technology contains at least about 0.005%, at least about 0.01 %, at least about 0.05%, at least about 0.08%, at least about 0.1%, at least about 0.2%, at least about 0.5% by weight of the composition of at least one fibroblast growth factor.
  • the unique biological gastropod fluid of the present technology also includes one or more enzymes.
  • the enzyme may include, for example, gelatinases, proteoglycanases, collagenases or combinations thereof.
  • Gelatinases include any proteolytic enzyme that allows a living organism to hydrolyse gelatin, including but not limited to, MMP2 and MMP9.
  • Proteoglycanases belong to the family of matrixmetalloproteinases (MMPs) which are zinc dependent endopeptidases. They are able to degrade all kinds of extracellular matrix proteins.
  • MMPs matrixmetalloproteinases
  • Suitable proteoglycanases include, but are not limited to MMP-23a, MMP-2, MMP23B, MMP-7, MMP-26.
  • Collegenases include any enzyme that break the peptide bonds of collagen. Collegenases are thought to assist in destroying extracellular structures in pathogens such as bacteria, and therefore are produced in response to tissue damage or an immune response.
  • the one or more enzymes comprise at least about 0.005%, alternatively at least about 0.01%, alternatively at least about 0.02%, alternatively at least about 0.05%, alternatively at least about 0.08%, or alternatively at least about 0.1 % by weight of the composition of the gastropod biological fluid.
  • the gastropod biological fluid also contains at least one oxygen carrier, e.g. hemocyanins, such as copper haemocyanin.
  • Hemocyanins are high molecular weight oxygen-carrying proteins. The oxygen-carrying proteins are thought to play a role in reversing oxidative damage and promoting cell proliferation and regeneration in skin cells.
  • the gastropod biological fluid contains at least about 0.001 %, alternatively at least about 0.01 %, alternatively at least about 0.02%, alternatively at least about 0.05%, alternatively at least about 0.08%, or alternatively at least about 0.1% of at least one oxygen carrier.
  • the gastropod biological fluid further includes at least one higher molecular weight protein.
  • Higher molecular weight proteins found in biological fluid may include mucin or mucus glycoproteins.
  • Mucin or mucus glycoproteins are a family of polydisperse molecules designed to carry out multiple tasks at mucosal surfaces throughout the body. Mucins are high molecular weight epithelial glycoproteins with a high content of clustered oligosaccharides O-glycosidically linked to tandem repeat peptides rich in threonine, serine and proline. They are rich in cysteine residues involved in sub unit crosslinking that form a macromolecular complex.
  • the gastropod biological fluid of the present technology comprises approximately at least about 0.001 %, alternatively at least about 0.005%, alternatively at least about 0.01 %, alternatively at least about 0.05%, or alternatively at least about 0.08% by weight of at least one high molecular weight protein.
  • the gastropod biological fluid further includes at least one trace element.
  • Trace elements include, for example, copper (Cu), zinc (Zn), iron (Fe), calcium (Ca 2+ ) or combinations thereof. Trace elements comprise about 0.0001%, alternatively at least about 0.0005%, alternatively at least about 0.001%, alternatively at least about 0.005%, or alternatively at least about 0.01% by weight of the biological fluid.
  • the gastropod biological fluid further may contain at least one high molecular weight antioxidant compound.
  • Antioxidants are molecules capable of slowing or preventing the oxidation of other molecules. Oxidation is a chemical reaction that transfers electrons from a substance to an oxidizing agent. Oxidation reactions can produce free radicals, which start chain reactions that damage cells. Antioxidants terminate these chain reactions by removing free radical intermediates, and inhibit other oxidation reactions by being oxidized themselves. As a result, antioxidants are often reducing agents, including but not limited to, thiols or polyphenols.
  • the gastropod biological fluid contains about 0.005%, alternatively at least about 0.008%, alternatively at least about 0.01%, alternatively at least about 0.05%, or alternatively at least about 0.1 % by weight of at least one higher molecular weight antioxidants.
  • cosmeceuticals, compositions or formulations comprising the gastropod biological fluid may also further comprise at least one additional antioxidant compound.
  • Any suitable naturally derived antioxidant compounds known in the art may be used, including but not limited to, olive oil derivatives, oleic acid, palmitic acid, uric acid, carotenoids, ubichinones, lipoic acid, vitamin C, vitamin E, phenoic compounds, resveratrol, beta carotene, selenium, high molecular weight antioxidants, low molecular weight antioxidant compounds, derivatives thereof and combinations there of.
  • the cosmeceutical, composition, or formulation may include the antioxidant compound at about 0.01% to about 10.0%, more suitably about 0.1% to about 5%, more suitably about 0.5% to about 3% by weight of the total composition.
  • the antioxidants may include about 0.01 %, about 0.05%, about 0.1 %, about 0.2%, about 0.5%, about 0.8%, about 1 .0%, about 1 .2%, about 1 .5%, about 1 .8%, about 2.0%, about 2.2%, about 2.5%, about 2.8%, about 3.0%, about 5.0%, about 7.0%, about 8.0%, about 10.0% by weight of the cosmeceutical, composition or formulation.
  • the gastropod biological fluid also contains about 0.005%, at least about
  • Low molecular weight antioxidant compounds are an important part of the antioxidative defense mechanisms of cells and organisms.
  • the main low molecular weight antioxidants include, but are not limited to, uric acid, ubichinones, lipoic acid, vitamins C and E, carotenoids, and phenolic compounds.
  • the gastropod biological fluid may be stored before usage and final formulation/composition/cosmeceutical processing.
  • the fluid may be rapidly frozen at between about -10 0 C to about -30° C, preferably aboil -15 ° C to about -20° C. Large amounts of the biological fluid may be stored for over a year by freeze drying.
  • the biological fluid is thermosensitive and can be denatured at about 60° C to 80° C, and the regenerative and antibiotic properties are lost if heated above about 90° C.
  • the biological fluid is stable in compositions when stored at temperatures between about -20° C to about 70° C.
  • the gastropod biological fluid may be used in an effective amount in a cosmeceutical.
  • a "cosmeceutical” is a composition comprising a combination of a cosmetic and biological and/or pharmaceutical product, such as an anti-wrinkle cream and a sunscreen.
  • the cosmeceutical of the present technology provides a beneficial property to the skin.
  • Beneficial properties include, but are not limited to, maintaining healthy skin; moisturizing; healing and preventing scarring; healing of sun damaged skin; preventing premature aging and wrinkles, inducing skin regeneration and wound healing; inducing skin repair; reducing oxidative stress; whitening skin pigmentation or lessening skin color; restoring tissue plasticity; opening clogged pores; treating ingrown hairs, razor bumps, dryness and photoaging; anti-inflammatory effects; reducing acne scarring; fighting acne breakouts; or fighting scalp and skin infections.
  • Suitable cosmeceuticals which may include gastropod biological fluid of the present technology preferably include anti-aging, anti-wrinkle, acne treatments, wound patches, wound salve, whitening compositions, and antimicrobial compositions.
  • An "effective amount" of the gastropod biological fluid is an amount that provides the desired beneficial outcome.
  • the effective amount of the gastropod biological fluid used in a cosmeceutical of the present technology depends on the delivery system being used and the type of application.
  • An effective amount of the gastropod biological fluid includes at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35% by weight of the cosmeceutical.
  • the gastropod biological fluid may be about 1%, about 2%, about 4%, about 6%, about 8%, about 10%, about 12%, about 14%, about 16%, about 18%, about 20%, about 22%, about 24%, about 25%, about 26%, about 28%, about 30%, about 32%, about 33%, about 34%, about 35% by weight of the cosmoceuctical by weight.
  • the cosmeceutical may include an effective amount of a gastropod biological fluid and a delivery vehicle capable of being applied to the skin.
  • Suitable delivery vehicles include, but are not limited to, a cream, a solution, a gel, an oil, a suspension, a dispersion, an ointment, a membrane, a patch, a powder, a capsule, a soap, a suppository, a derivative thereof or a combination thereof.
  • the amount of gastropod biological fluid may depend on the delivery vehicle being used.
  • a cream used for treatment of the face, hands or body may include about 15% to about 20% gastropod biological fluid.
  • the composition, cosmeceutical, or formulation may contain about 1 % to about 5% gastropod biological fluid.
  • the composition, cosmeceutical, or formulation may include about 30% to about 35% gastropod biological fluid.
  • the composition, cosmeceutical, or formulation may be a powder to be applied to a would area or specific skin areas containing about 15% to about 20% of the gastropod biological fluid.
  • the composition, cosmeceutical, or formulation may be a mask to cover a large wound or a face mask that includes about 20% to about 25% gastropod biological fluid.
  • the cosmeceutical may be a membrane to cover part of the face or body area or a wound area that includes about 20% to about 25% gastropod biological fluid.
  • the cosmeceutical may include about 15% to about 25% gastropod biological fluid.
  • the cosmeceutical may include a pill or a capsule for oral administration of the active ingredients that includes about 25% to about 30% by weight of the total cosmeceutical of the gastropod biological fluid.
  • the cosmeceutical may be a soap to wash hand, face or body which includes the gastropod biological fluid at about 1 % to about 10%, more suitably about 1 % to about 3% by weight of the total composition.
  • the cosmeceutical may further include at least one olive oil or olive oil derivative.
  • Olive oil derivatives include, but are not limited to, oleic acid, palmitic acid, and other fatty acids, squalene, sterols including phytosterol and tocosterols, tyrosol, hydroxytryrosol including oleocanthal and oleuropein, Peg-4 olivat ⁇ , surfactant sorbitan olivate, derivatives thereof, or combinations thereof.
  • Olive oil derivatives contain potent antioxidant properties along with moisturizing and immune stimulating properties.
  • the compositions include about 0.01% to about 10%, more suitable about 0.5% to about 5% by weight of at least one olive oil derivative.
  • the at least one olive oil derivate may be about 0.08%, about 0.1%, about 0.2%, about 0.3%, about 0.5%, about 0.8%, about 1 .0%, about 1 .2%, about 1 .5%, about 1 .8%, about 2.0%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.8%, about 3.0%, about 3.2%, about 3.4%, about 3.6%, about 4.0%, about 4.2%, about 4.5%, about 4.8%, about 5.0%, about 7.0%, about 10.0% of the total weight of the cosmeceutical, composition, or formulation of the present invention.
  • the olive oil derivative may suitably be combination of peg 4 olivate, squalane, and sorbitan olivate.
  • the at least one olive oil derivative includes about 0.2% to about 2.0%, more suitably 0.8% to about 1.0% peg 4 olivate; about 0.2% to about 2.0%, more suitably about 0.8% to about 1 .0% squalane; and about 0.2% to about 2.0%, more suitably about 0.8% to about 1 .0% sorbitan olivate.
  • the cosmeceutical further includes squalane at about 0.01 % to about 10%, more suitably about 0.1 % to about 2%, more suitably about 0.8% to about 1 .2% based on the total weight of the cosmeceutical, composition, or formulation.
  • Squalane is a natural organic compound obtained from mainly from shark liver oil or olive oil, but also found in botanic sources such as amaranth seed, rice bran, wheat germ, and olives.
  • Squalene is used in the cosmeceutical as a natural moisturizer and antioxidant which penetrates skin quickly and does not leave a greasy residue.
  • the cosmeceutical further includes hyaluronic acid.
  • Hyaluronic acid may comprise about 0.01% to about 10%, more suitably about 0.1% to about 5%, more suitably about 0.5 to about 2%, more suitably about 0.8% to about 1 .0% of the total weight of the composition.
  • Hyaluronic acid is a non-sufated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues. It is one of the main components of extracellular matrix and contributes to cell proliferation and migration, participates in a number of cell surface receptor interactions and is involved in tissue repair.
  • Hyaluronic acid has been shown to be an effective scavenger of free radicals and may also be effective when scavenging oxidants caused by ultraviolet radiation.
  • the hyaluronic acid may play a role in tissue healing and reducing oxidative stress on the skin.
  • hyaluronic acid and squalene are included with the gastropod biological fluid to form the desired cosmeceutical.
  • the cosmeceutical, composition or formulation may include a preservative.
  • the preservative is an ingredient added to the cosmeceutical, composition, or formulation to prevent the growth of microorganisms including bacteria, viruses, yeast and fungi, and to protect the cosmeceutical from damage caused from air and light.
  • the cosmeceutical may include the preservative at about 0.01 to about 10%, more suitable about 0.1 to about 5%, more suitably about 0.5% to about 5%, more suitably about 0.8% to about 1 .2% based on the total weight of the composition.
  • Particularly suitably preservative includes phenoxyethanol, ehtylhexylglycerin, derivatives thereof, or combinations thereof.
  • Phenoxyethanol is an organic chemical compound, a glycol ether bactericide.
  • Ethylhexylglycerin (1 -(2- ethylhexyl)-glycerinether) is an enhancer of the action of preservatives, inhibits bacterial growth and is a mild humectant and skin emollient.
  • Any suitable preservative known in the art to be used in cosmetics may be used.
  • Other alternative preservatives include paraben.
  • the preservative may be a combination of botanical extracts.
  • Suitable botanical extracts include, for example, organum vulgare leaf extract, thymus vulgaris extract, cinnamomum zeylanicum bark extract, rosmarinus officinalis leaf extract, lavandula augustifolia flower extract, citrus medica limonum peel extract, menthe piperita leaf extract, hydrastis Canadensis root extract, olea europaea leaf extract, derivatives thereof, or a combination thereof.
  • the cosmeceutical, composition or formulation may further comprises at least one solvent.
  • a solvent is a liquid that dissolves a solid, liquid or gas into a resulting solution. Suitable solvents include, but are not limited to water, glycerin, alcohols, derivatives thereof and combinations thereof. Glycerin is a colorless, dense sugar alcohol that can also dissolve easily in alcohol or water. Glycerin also is highy "hygroscopic" which means that it absorbs water from the air. Glycerin provides the compositions with means of improving smoothness, providing lubrication and as a humectant. The amount of glycerin depends on the delivery system of the cosmeceutical, composition or formulation.
  • the cosmeceutical, composition or formulation includes the solvent at about 0.1% to about 99%, alternatively about 10% to about 90% of the total weight.
  • the cosmeceutical may include the solvent at least about 0.1%, about 1 .0%, about 5%, about 8%, about 10.0%, about 12.0%, about 15.0%, about 18.0%, about 20.0%, about 22.0%, about 24.0%, about 25.0%, about 28.0%, about 30.0%, about 32.0%, about 35.0%, about 36.0%, about 38.0%, about
  • composition 94.0%, about 96.0%, about 98.0% by weight of the composition, cosmoceutical or formulation.
  • the cosmeceutical, composition or formulation may include an abrasive agent.
  • abrasive agents provide a sloughing outcome of dead cells or tissue and other debris without causing damage to the skin.
  • Suitable abrasive agents are known in the art and include, but are not limited to, corudrum oxide.
  • the cosmeceutical, composition or formulation may include about 0.0005% to about 0.05%, more suitably about 0.001 % to about 0.01 % by weight of the composition.
  • the cosmeceutical, composition or formulation may include an aromatic essence.
  • An aromatic essence may provide an aroma to the formulation, which include, but are not limited to, for example, vanilla, jasmine, violet, rose, lavender, chamomile, peach, strawberry, coconut, green tea, mint, citrus, etc.
  • the cosmeceutical, composition or formulation may include the aromatic essence at about 0.005%, alternatively at about 0.01 %, alternatively at about 0.05%, alternatively at about 0.1 %, alternatively at about 0.5%, alternatively at about 0.8%, alternatively at about 1 .0%, or alternatively at about 2.0%.
  • the cosmeceutical, composition or formulation may includes about 5.0% to about 10.0% stearic acid, about 0.1% to about 1 % single stearic acid glyceride, about 1 % to about 10% of at least one oil, about 0.1% to about 1 % octadecanol, about 5% to about 15% glycerin, about 10% to about 20% gastropod biological fluid, about 0.1% to about 1 .0% potassium hydoxide, about 0.1% to about 2.0% fragrance essence, and about 0.1 % to about 2.0% of a preservative comprising a combination of phenoxyethanol and ethylhexygycerin.
  • the one or more oil may include white oil, silicon oil, a derivative thereof, or a combination thereof.
  • the composition, cosmeceutical, or formulation includes at least one oil containing about 4.0% to about 5.0% white oil and about 1 .0% to about 2.0% silicon oil based on total weight of the composition.
  • This cosmeceutical may further comprise water in an amount sufficient to bring the composition to approximately 1000 grams.
  • the cosmeceutical, composition, or formulation comprises about 2.4% to about 3.0% of at least one olive oil derivative, about 10% to about 15% gastropod biological fluid, about 0.8% to about 1.2% hyaluronic acid, about 10.0% to about 15.0% glycerin, and about 0.8% to 1.2% a preservative (e.g., phenoxyethanol and ethylhexygycerin) based on total weight of the composition.
  • a preservative e.g., phenoxyethanol and ethylhexygycerin
  • these embodiments include water to bring the weight of the formulation to approximately 100Og.
  • the cosmeceutical, composition or formulation may further comprises about 0.5% to about 5.0%, more suitably about 1.5% to about 2.5% seaweed extract based on total weight of the composition. Seaweed extract provides oligoelements that promote cell growth.
  • the cosmeceutical further comprises about 0.01% to about 10.0%, more suitably about 0.1% to about 2.0% hyaluronic acid, about 0.1 % to about 10%, more suitably about 1.0% to about 4% chamomile.
  • Chamomile provides anti-inflammatory properties and is useful for sensitive skin.
  • the cosmeceutical, composition or formulation may include about 0.1% to about 10%, more suitably about 2.0% to about 5.0% saccharide isomerate based on total weight of the composition.
  • Saccharide isomerate includes a carbohydrate complex similar to complexes found in the skin that promotes the skin to retain water.
  • the formulation may include licorice extract (Glycyrrhiza inflata extract) which has antioxidant, anti-inflammatory, anti-irritant, antimicrobial, and sebum regulator activities.
  • the licorice extract may include about 0.5% to about 10%, more suitably about 1 .0% to about 5.0% based on the total weight of the composition.
  • the composition may include about 0.1 % to about 10%, more suitably about 1 % to about 5% based on total weight of the composition of a preservative derived from natural botanical extracts. These botanical extracts provide antimicrobial, antibacterial, antifungal, anti-yeast and/or antiviral properties.
  • Suitably botanical extracts include, but are not limited to Origanum Vulgare (Oregano) Leaf Extract, Thymus Vulgaris (Thyme) Extract, Cinnamomum Zeylanicum (Cinnamon) Bark Extract, Rosmarinus Officinalis (Rosemary) Leaf Extract, Lavandula Augustifolia (Lavender) Flower Extract, Citrus Medica Limonum (Lemon) Peel Extract, Mentha Piperita (Peppermint) Leaf Extract, Hydrastis Canadensis (Golden Seal) Root Extract, and Olea Europaea (Olive) Leaf Extract.
  • Origanum Vulgare (Oregano) Leaf Extract Thymus Vulgaris (Thyme) Extract
  • Cinnamomum Zeylanicum (Cinnamon) Bark Extract Rosmarinus Officinalis (Rosemary) Leaf Extract
  • Lavandula Augustifolia (Lavender) Peel Extract Citrus Medica Limonum (Lemon) Peel Extract
  • the cosmeceutical, composition or formulation of the present technology may also include an effective amount of a gastropod biological system and ingredients derived from the traditional Chinese medicine to create novel formulations.
  • suitable ingredients derived from Chinese medicine include, but are not limited to, oils (e.g., rose oil, emulsion aburagi fruit oil, corn embryo oil, wheat embryo oil, fibert oil, sesame oil), sugars and sugar derivatives(e.g. dextrose, AHA, glycolic acid, glycerins), plant and fruit essence (e.g.
  • Ginko essence, tulip essence, aloe, hamamelis essence, thyme essence, salicylic acid, willow bark essence, essence of carbamide and liquorice, citrus essence, blackberry essence); herbs (e.g. Ganoderma Lucidum, Panax Ginseng); fruits, nuts, vegetable and derivatives thereof (e.g.
  • Sodium C14-16 olefin sulfate C14-16, triethanolamine, honey essence, levorotation-C, amides, jobs of tears, arginine, lactic acid, urea, etc. may be used for a variety of skin care treatment products, including but not limited to, wound patches, postoperative healing membranes, rejuvenation essence, antiphlogistics (purification conditioning cream), anti-acne cream, anti-wrinkle cream, moisten facial cleansing soap, shower gel, hand cream, whiten cream, post-partum dressing, hemorrhoids thrombolysis, anti-ultraviolet skin care cream, anti-drying skin-care cream, anti-burn healing cream).
  • skin care treatment products including but not limited to, wound patches, postoperative healing membranes, rejuvenation essence, antiphlogistics (purification conditioning cream), anti-acne cream, anti-wrinkle cream, moisten facial cleansing soap, shower gel, hand cream, whiten cream, post-partum dressing, hemorrhoids
  • a particular embodiment of a whitening cream includes the gastropod biological fluid, vitamin C, yellow gentian grass, kojic acid, at least one antioxidant (e.g., green tea),and at least one plant extract.
  • Suitable plant extracts include sorbic, Occident pear, olive fruit, gingko, ursolic, foreign beech, seaweed, birch, mulberry, Panax, etc.
  • a particular embodiment of a sunscreen cosmeceutical cream includes the gastropod biological fluid, and at least one UV-blocking agent selected from the group comprising PABA (e.g., Para-aminobenzonic acid, Padimate O, Padimate A, Glyceryl PAA), salicylates (e.g., homosalate, octyl salicylate, trolamine salicylate), cinnamates (e.g., octyl methoxycinnamate, cinoxate); benzophenones (e.g., oxybenzone, dioxybenzone, sulisobenzone), anthranilate, methylanthranilate, physical agents (e.g. xinc oxide 4, red petroleum) and a combination thereof.
  • PABA e.g., Para-aminobenzonic acid, Padimate O, Padimate A, Glyceryl PAA
  • salicylates e.g., homosalate, oc
  • a particular embodiment of a therapeutic acne removing cosmeceutical cream of the present technology includes, for example, the gastropod biological fluid, gentian garlic extract, hop oil, chamomile extract, sunflower extract, tea-tree oil, and calendula.
  • Suitable additional components include, for example, propylene glycol, polypropylene glycol, butylene glycol, hexylene glycol, caprylic, capric, lauric triglycerides, cyclomethicone, dimethicone, fatty acids, hyaluronic acid, sodium PCA, collagen, ceramide, elastin, lecithin, derivatives thereof or combinations thereof.
  • the cosmeceuitcal may also be prepared as a therapeutic cream as described in the following examples.
  • a therapeutic cream may be use to (i) treat types of burns including thermal and chemical burns, (ii) prevent or treat radiodermatitis, and (iii) to prevent or treat sunburn.
  • the cosmeceutical of the present technology may be a nourishing face or hand cream for use on normal, dry or oily skin. It is believed that the gastropod biological fluid repairs the epithelium and rejuvenates the skin. It is also believed that the cream also serves as an exfoliate, eliminating the outer dead skin layer (epithelial corneal layer) and encouraging the growth of new skin.
  • the use of panthotenic acid and heparin in the cream is optional, but preferred because they increase the restorative and penetrating actions.
  • the cream tones and revitalizes the skin and may be used for treatment of rosacea, eczema, dermatitis and very dry skin conditions.
  • a cosmeceutical of the present technology may be a clarifying and moisturizing liquid for dry, normal or oily skin.
  • the resulting liquid cleanses and smoothes the skin and heightens the transparency of the complexion. It serves also as a base for make-up (e.g., powder) and protects the skin from the harmful action of cosmetic powders which consist of minerals which have a hardness value of 1 on the Mohr scale.
  • a suitablle formulation comprises about 2 ml of gastropod biological fluid in about 200 g total product.
  • the clarifying and moisturizing liquid preferably includes about 0.5%, alternatively about 1 .0%, alternatively about 2%, alternatively about 5%, alternatively about 10%, alternatively about 15% of the gastropod biological fluid of the present invention.
  • a cosmeceutical of the present technology may be a toning and cleansing liquid.
  • the liquid is recommended for use on delicate skin or skin where acne is present. It produces an intense refreshing feeling and a soothing, restful sensation
  • a cosmeceutical of the present technology may be an anti-wrinkle cream for daily or weekly use.
  • the cream is easy to apply and best if used only on areas where wrinkles exist.
  • the gastropod biological fluid active ingredient is believed to be effective in reducing wrinkling, whereas the other components produce an astringent action. The effect of the cream lasts at least one week after each application.
  • An excipient specially designed for the preservation of biological substances and for use in dermatological and cosmetic treatments is used in YoungerTM Biocream commercially available from Pharmacom located in Iowa City, Iowa.
  • This preferred excipient is made from a polyethylene glycol mixture having a molecular weight range of about 900 to about 1300, cetyl alcohol, glyceryl monostearate, selected mineral oils, calcium carbonate in micropowder form, zinc oxide in micropowder form, and an aqueous phase.
  • the amount of the aqueous base should be an amount which is sufficient to give the composition of the desired consistency. If the amount of the aqueous base is above 35%, a preservative should be used.
  • the aqueous base comprises water, a saline solution [e.g., a solution of 0.9% sodium chloride which contains no pyrogenous matter], or physiological serum. The percentages are by weight and total 100%. The percentages can be easily adjusted with minimal experimentation to provide the desired creamy consistency.
  • the excipient is odorless and white and unaffected by ultraviolet radiation, x-rays, and gamma rays. It can be used in the preparation of creams, milks, pomades and the like.
  • the excipient promotes the absorption of the active ingredient through the skin, does not break down between about -20 ° C and about +70 ° C, and is unaffected by humidity, and thus may be used in many climatic extremes. Further, the excipient is stable and not affected by decimeter waves, ultraviolet radiation, gamma radiation of 1 .3 MeV, and photons. Preferably, is also unaffected by the container in which it is stored. The importance of this excipient is not based on exotic components, but on their proportions which is the result of the numerous experimental formulations.
  • Optional ingredients which can be used in the cosmeceuticals for the cases where the aqueous base is greater than 35% include a preservative such as a methyl paraben, an oxidation inhibitor such as sodium bisulfite, and other ingredients typically included in therapeutic compositions.
  • Optional ingredients which can be used in the compositions, cosmeceuticals, or formulations of the present technology include mineral oils such as paraffin, turtle, or other oils in amounts of about 1% to about 10%, preferably about 2%; boric acid in amounts of about 0.1 % to about 0.6%; preferably about 0.3%; chlorophyll in amounts of about 0.25% to about 1 .5%, preferably about 1 .0%; formol (e.g., about 30% concentration) in amounts of about 0.3% to about 3%, preferably about 0.4%; glycerin in amounts of about 0.1 % to about 4.0%, preferably about 0.5%; heparin in amounts of about 0.01 % to about 0.4%, preferably about 0.1%; lanolin in amounts of about 0.1 % to about 6%, preferably about 0.5%; menthol in amounts of about 0.25 to about 2.5%, preferably about 1 %; panthotenic acid in amounts of about 0.01 % to about 0.5%, preferably about 0.1%; potassium aluminum s
  • Helix Aspersa M ⁇ ller originated from France and were grown on protein- rich food. Helix Aspersa M ⁇ ller snails were centrifuged at 2G for 10 minutes with 3 pulsations. A pulsation was performed by accelerating the centrifuge to 2G, decelerating the centrifuge and re-accelerating the centrifuge to 2G. During each pulsation, the centrifuge was accelerated to 2G, decelerated and re-accelerated to 2G. After centrifugation, the fluid secretion of the snail was collected by use of a straw for removing the fluid from the snail and collected into a centrifuge-compatible tube. The fluid secretion was centrifuged at 2000 rpm for 10 minutes to remove any large particles.
  • the supernatant was decanted into a cylinder containing a 0.1 ⁇ m Millipore filter.
  • the cylinder was hermetically sealed with a closure bearing a connection to a compressed air system to facilitate filtration.
  • the filtrate was then used as the gastropod biological fluid of the presently described technology.
  • Aspersa M ⁇ ller snails were processed to determine their GAG (glycosaminoglycan) concentration.
  • the shell of the snail was removed, and the whole soft body was defatted using three, 24-hour extractions with acetone.
  • the fat-free dried snail was cut into a fine powder using a razor blade.
  • Approximately 4 g of dried, defatted, pulverized powder was suspended into 40 ml of 0.05 M sodium carbonated buffer (pH 9.2). The suspension was shaken for 48 hours at 200 rpm at 60 ° C after adding 2 ml of Alcalase (2.4 Anson units/g).
  • the digestion mixture was cooled to 4 °C, and trichloroacetic acid was added to a final concentration of 5%.
  • the capillary (75- ⁇ m inner diameter, 375- ⁇ m outer diameter, 68 cm long) was manually washed before use with 0.5 ml of 0.5 M sodium hydroxide followed by 0.5 ml of distilled water, and then 0.5 ml running buffer. Samples were applied using gravity injection (20 seconds) by hydrostatic pressure (45 mm), resulting in a sample volume of 9.2 ml. Each experiment was conducted at a constant 18,000 V. Data collection was at 232 nm. Peaks were identified by co-injection with disaccharide and oligosaccharide standards prepared and characterized in our laboratory.
  • the size of products formed on treating gastropod GAG with heparin lyase Il was determined by gel permeation chromatography on a Sephadex G-50 (superfine) column (1 .5 ⁇ 25 cm) eluted with 0.2 M sodium chloride and monitored using a carbazole assay.
  • the disaccharide product of heparin lyase Il treatment was prepared for analysis by semipreparative strong anion exchange HPLC and desalted by chromatography on a 5- cm x ⁇ .5-m Bio-Gel P-2 column.
  • the reactions to show acid glycosaminoglycans revealed five types of mucous cells in the mantle: cells with glycoproteins only, with sulphated acidic proteoglycans, with proteoglycans containing carboxyl groups, and with glycoproteins and proteoglycans containing carboxyl groups (see e.g., Fig. 2-3).
  • Some mucous cells of the foot epithelium contained carboxyl acidic proteoglycans (see e.g., Fig. 2-4) others also glycoproteins (see e.g., Fig. 2-5). Sialic acid and hyaluronic acid were not found in the tegument of the mantle and foot.
  • Example 3 Gastropod biological fluid Induces Fibroblast Proliferation in vitro and Regulates Fibroblast Cytoskeleton Reorganization. [0090] To investigate if the regenerative properties of gastropod biological fluid
  • Fibronectin was stained with the anti-80 kDa polyclonal antibody followed by incubation with Alexa488-conjugated anti-rabbit antibody. Samples were mounted in Mowiol, and were examined in a Leica DMR photomicroscope with 63x and 100x immersion objectives. Images were processed using Leica QFish software and results are shown in Fig. 4. Gastropod biological fluid from Helix Aspersa M ⁇ ller induced fibronectin assembly compared to control cells. Interestingly, biological fluid appeared to increase fibronectin secretion since it increased fibronectin deposition by the cells in the absence of exogenous fibronectin.
  • GST Glutathione-S-Transferase
  • SOD Superoxide dismutase
  • Gastropod biological fluid filtrate (about 100 ml to about 150ml)
  • Glycerin (about 1 10g to about 13Og)
  • Corundum oxide (about 4g to about 6g)
  • Sorbitan Olivate from Olive Oil (about 10g)
  • Sorbitan Olivate from Olive Oil (about 1 Og)
  • Gastropod biological fluid (about 100 ml)
  • Aromatic Essence (5 ml) Phenoxyethanol & Ethylhexygycerin (about 1 Og)
  • Distilled water (about 625 ml) plus enough water to bring the weight of the product to about 100Og.
  • Aromatic Essence (about 5 ml)
  • Distilled water (about 600 ml) plus enough water to bring the weight of the product to 100Og.
  • Rose Hip Seed Oil (about 8 to about 10 ml) Gastropod biological fluid filtrate (about 100 to about 150 ml) Olive Oil emulsifier (about 8 to abouti Og) Seaweed Extract (about 18g to about 22g) Hyaluronic Acid (about 8g to about 1 Og) Chamomile (about 28g to about 32g) Saccharide lsomerate (about 38g to about 42g), Licorice (Glycyrrhiza inflata) Extract (about 22g to about 25g)
  • a preservative (about 18g to about 22g): made with the following botanical extracts in minute quantity: Origanum Vulgare (Oregano) Leaf Extract, Thymus Vulgaris (Thyme) Extract, Cinnamomum Zeylanicum (Cinnamon) Bark Extract, Rosmarinus Officinalis (Rosemary) Leaf Extract, Lavandula Augustifolia (Lavender) Flower Extract, Citrus Medica Limonum (Lemon) Peel Extract, Mentha Piperita (Peppermint) Leaf Extract, Hydrastis Canadensis (Golden Seal) Root Extract, and Olea Europaea (Olive) Leaf Extract
  • Distilled water (about 600 ml) plus enough water to bring the weight of the product to about 100Og.
  • a preservative ( about 2Og): made with the following botanical extracts in minute quantity: Origanum Vulgare (Oregano) Leaf Extract, Thymus Vulgaris (Thyme) Extract, Cinnamomum Zeylanicum (Cinnamon) Bark Extract, Rosmarinus Officinalis (Rosemary) Leaf Extract, Lavandula Augustifolia (Lavender) Flower Extract, Citrus Medica Limonum (Lemon) Peel Extract, Mentha Piperita (Peppermint) Leaf Extract, Hydrastis Canadensis (Golden Seal) Root Extract, and Olea Europaea (Olive) Leaf Extract
  • an excipient preferred for use in therapeutic compositions of the present technology containing the gastropod biological fluid of the present technology was made by the following process: Cetyl alcohol (15 g.) was fused by standard processes and then 50 g of a mixture of polyethylene glycols [polyethylene glycol 400 (49.%), 1500 (1 .0%) and 4000 (49.5%)] was added to form a first mixture. When the fusion point was reached, this mixture was transferred to a commercial pharmaceutical laboratory. A second mixture was prepared by fusing 5 g. of glycerol monostearate and adding 12.6 g (12.6 ml) of distilled water or preferably a physiological serum previously heated to 80 ° C.
  • the second mixture was added to the first mixture.
  • 7 g of calcium carbonate and 0.4 g of zinc oxide, both in the form of micropowders, and 10 ml of paraffin oil were added to the mixture.
  • a total of about 25 to about 30 ml of the gastropod biological fluid (or about 25 to about 30 g of the freeze-dried fluid) was mixed with about 500 to about 600 g of the excipient described above at about 30 ° C to about 40 ° C in a mixer.
  • a total of about 500 to about 700 g of cream was obtained.
  • Example 10 Preparation of a nourishing hand cream
  • the cream was prepared by mixing about " ! 00 g total, 5 g of the gastropod biological fluid, 4 cc. of glycerin, and 0.5 g of lanolin.
  • the resulting cream was very smooth and it spread well. It can be applied in the day or at night to regenerate, rejuvenate, and smooth the skin.
  • Example 11 Preparation of a clarifying and moisturizing liquid
  • This example describes the preparation of a clarifying and moisturizing liquid for dry, normal or oily skin.
  • the liquid was prepared by mixing 200 g. of the excipient, 20 cc. of the gastropod biological fluid, 0.3 g of potassium aluminum sulfate, and 10 c. of rosewater.
  • the liquid was prepared as above except that 2 cc. of a mineral oil, e.g., paraffin oil, was added. Colorants may optionally be added.
  • the resulting liquid cleanses and smoothes the skin and heightens the transparency of the complexion. It also serves as a base for make-up (e.g., powder). Finally, it protects the skin from the harmful action of cosmetic powders which consist of minerals which have a hardness value of 1 on the Mohr scale.
  • the moisturizing kinetic study was extended to 21 days.
  • the study participants applied the formulations (10% gastropod biological fluid or Placebo) twice a day to their skin.
  • the moisture level was measured on day 22 for each study participant and the results averaged as shown in Fig. 8.
  • the moisturizing level of the gastropod biological fluid (snail serum) on the treated area on day 22 was clearly higher (+12.5%) as compared with the initial values of each volunteer before the start of the study (D0/T0).
  • Example 12 Preparation of a toning and cleansing liquid
  • the liquid was prepared by mixing 100 g total, 10 cc. of the gastropod biological liquid, 100 cc. of physiological serum, 0.3 g. of potassium aluminum sulfate, 0.5 g. of boric acid, and 0.1 g. of copper sulfate.
  • the liquid is recommended for use on delicate skin or skin where acne is present. It produces an intense refreshing feeling and a soothing, restful sensation.
  • An anti-wrinkle cream for daily use was prepared by mixing 250 g total, 5 cc. of the gastropod biological fluid, 7 g. of lanolin, and 1 .56 g. of potassium aluminum sulfate can be added to the base excipient or a physiological serum.
  • a concentrated cream for use no more often than once a week is prepared as above except that 15 g. of the active principal, 15 g. of lanolin, and 2 g. of potassium aluminum sulfate are used.
  • the cream is easy to apply and is used only on areas where wrinkles exist.
  • the active ingredient is effective in reducing wrinkling, whereas the other components produce an astringent action.
  • each pixel is composed of two adjacent metal plates.
  • the gray level values of the output image represent the 3D skin sensor gives a map of the skin surface by modulating the gray level values of each pixel in function of the capacitance measured, which is in turn a function of the distance between the sensor plate and the 3D profile of the skin surface.
  • the first part of experiments is to measure planar distances between well identified topographic structures. We paid our attention on wrinkles, identified by high gray level values with respect to the surrounding skin, which in contrast appear darker in the capacitive images.
  • a preprocessing step is performed with the aim of reducing misleading factors described above.
  • wrinkle's profiles which are transformed in an analytical form through using natural cubic spline.
  • what we measure is the inter-wrinkle distance (hereinafter, brief Iy IWD), that is the distance, in micron, between axes passing by two following local minima extracted from the 2D profile and belonging to adjacent wrinkles.
  • the second part of the experiments is devoted to validate these measures by comparing them with measures of the same body site area achieved through profilometric analysis.
  • Silicone skin replicas have been analyzed with an optical profilometer equipped with a 20 ⁇ magnification objective and a .5 ⁇ scale factor reducer lens, thus giving an overall magnification factor of 10 ⁇ .
  • the system based on white light interferometry, provides accurate surface data depths of a limited region of the replica, being the field of view of 0.62 x 0.47mm2 and 640 x 480 pixel the image resolution.
  • the concentrated cream was tested on more than 200 healthy woman volunteers.
  • the group was divided and tested with anti-wrinkle cream describe above or placebo for 30 days.
  • the anti-wrinkle effect lasted for approximately one week after each application.
  • the depth of wrinkles was diminished after 15 and 30 days for skin treated with the anti-wrinkle cream (Fig. 9E and F) as compared with placebo (Fig. 9B and C). This observation that the depth of the wrinkle is significantly decreased after 30 days of treatment confirms the anti-wrinkle benefits of cream containing the gastropod biological fluid of the present technology.
  • a sunscreen cream was prepared by mixing 250 g total with 5 ml of the gastropod biological fluid filtrate.
  • a sunscreen tanning cream is prepared as above except that 1 g of metallic iodine and 0.3 g of Vitamin C are added.
  • 1 g of metallic iodine and 0.3 g of Vitamin C are added.
  • one thin application of the sunscreen before exposure to the sun is sufficient. If one has suffered sunburn, a thin coating of the cream may be applied every six hours.
  • the sunscreen cream was used in the treatment of 306 fair-complexioned test subjects.
  • the products were used preventively in 6% of the cases and curatively in 94% of the cases.
  • the sunscreen was used in treating test subjects who suffered sunburns at swimming pools with highly chlorinated water, where the subjects showed blistering (phlytena), and persons who suffered sunburns at beaches.
  • the cream eliminated the pain and burning sensation within seconds of being applied, blisters disappeared in 5 to 6 hours; and the sunburn was gone in 24 hours.
  • Ultraviolet irradiation causes adverse effects like sunburn, photosensitivity reactions or immunologic suppression.
  • the aim of this study was to evaluate the photo- protective outcome of Pharmacom TMYounger Sunscreen Cream by the determination of erythema indexes and the assessment of the high molecular weight antioxidants and its metabolites in human dermis. These substances were used as markers of oxidative effect.
  • Eight healthy female subjects were enrolled in this study. Two abdominal areas were exposed to solar simulated irradiation with three minimal erythema doses, one with the Sunscreen Cream application and the other site without the Sunscreen Cream application. Two other areas were used as control, one without SPF8 application and the other site after SPF8 application.
  • Ascorbic acid and its metabolites were collected from human dermis by microdialysis and assessed by gas chromatography mass spectrometry. Irradiated site without sunscreen application had significantly demonstrated lower dermis ascorbic acid concentrations and a higher erythema index than the three other sites (P ⁇ 0.05). Threonic acid, oxalic acid and xylose dermis concentrations were significantly higher in site III than in the control site I (P ⁇ 0.05). The protected-irradiated site did not show erythema formation and there was stability of ascorbic acid dermis concentrations with non-variation in its metabolites. These results suggest that sunscreen compositions of the present technology may act as a sunscreen and prevent oxidative and UV damage to the skin.

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Abstract

L'invention concerne de manière générale des compositions de soins de la peau, des produits cosmétiques ou des formulations et des procédés pour les fabriquer ou les utiliser. Plus spécifiquement, la présente invention concerne de manière générale des procédés de fabrication et d'utilisation de compositions, de produits cosmétiques ou de formulations comprenant une composition unique collectée et raffinée à partir d'un gastéropode, à savoir Helix Aspersa Mller, entre autres.
PCT/US2008/068047 2007-06-23 2008-06-24 Fluide biologique de gastéropode, procédé de fabrication et de raffinage, et utilisation WO2009002982A2 (fr)

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US12/643,751 US20100233111A1 (en) 2008-04-06 2009-12-21 Gastropod biological fluid, method of making and refining and use

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Cited By (7)

* Cited by examiner, † Cited by third party
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WO2012097466A1 (fr) * 2011-01-21 2012-07-26 Comercializadora Muciderm Ltda. Composition pharmaceutique et dispositif pour prévenir, traiter et soigner des ulcères du pied diabétique et autres plaies, à base de bave d'escargot de l'espèce cryptophalus aspersus ou helix aspersa müller et d'excipients et/ou d'additifs acceptables sur le plan pharmaceutique
WO2016029328A1 (fr) * 2014-08-25 2016-03-03 Muciderm S.A Composition pharmaceutique pour la prévention, le traitement et la suppression du psoriasis, contenant de la bave d'escargot, de la camomille et du miel
WO2016054757A1 (fr) * 2014-10-10 2016-04-14 Muciderm S.A. Composition pharmaceutique pour la prévention, le traitement et la suppression de la rosacee, contenant de la bave d'escargot, de la camomille et du miel
EP3124030A1 (fr) * 2015-07-30 2017-02-01 Aristotle University Of Thessaloniki-Research Committee Procédé de production d'un extrait d'escargot petit-gris (cornu aspersum) et ledit extrait
IT201700122874A1 (it) * 2017-10-31 2019-05-01 Roberto Leone Composizione edibile a base di bava di lumaca per la conservazione degli alimenti e per la sanificazione di superfici, ambienti e strumenti
CN110314247A (zh) * 2019-08-02 2019-10-11 深圳国佳产业基金管理有限公司 蜗牛精华提取物活性敷料及其制备方法
WO2022150018A1 (fr) * 2021-01-08 2022-07-14 Aden International Co.,Ltd. Nouveau procédé d'isolement, de purification et de caractérisation de substances semblables à l'héparine provenant de mucus d'escargot (achatina fulica) et utilisations associées

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CN102079979B (zh) * 2010-12-08 2013-04-24 华南农业大学 角鲨烯与β-胡萝卜素的组合物及其在卷烟中的应用
CN104975001A (zh) * 2015-07-15 2015-10-14 中国海洋大学 一种无损伤提取脉红螺基因组dna的方法
CN111679000A (zh) * 2020-05-29 2020-09-18 费森尤斯卡比华瑞制药有限公司 检测肠内或肠外营养制剂中维生素c杂质的方法

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US6946551B2 (en) * 2003-03-12 2005-09-20 New Life Resources, Llc Preparation of hyaluronic acid from eggshell membrane

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US3899006A (en) * 1971-03-29 1975-08-12 Pneumatiques Caoutchouc Mfg Tubes provided with connecting flanges
US5538740A (en) * 1991-03-01 1996-07-23 Atherton Investments, Ltd. Therapeutic and cosmetic compositions for treatment of skin
US6946551B2 (en) * 2003-03-12 2005-09-20 New Life Resources, Llc Preparation of hyaluronic acid from eggshell membrane

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012097466A1 (fr) * 2011-01-21 2012-07-26 Comercializadora Muciderm Ltda. Composition pharmaceutique et dispositif pour prévenir, traiter et soigner des ulcères du pied diabétique et autres plaies, à base de bave d'escargot de l'espèce cryptophalus aspersus ou helix aspersa müller et d'excipients et/ou d'additifs acceptables sur le plan pharmaceutique
US20130309296A1 (en) * 2011-01-21 2013-11-21 Elmo Ernesto Moreno González Pharmaceutical composition and device for preventing, treating and curing ulcers on a diabetic foot and other wounds, which includes snail slime from the species cryptophalus aspersus or helix aspersa muller and pharmaceutically acceptable carriers and/or additives
US9125964B2 (en) * 2011-01-21 2015-09-08 Muciderm S.A. Pharmaceutical composition and device for preventing, treating and curing ulcers on a diabetic foot and other wounds, which includes snail slime from the species Cryptophalus aspersus or Helix aspersa muller and pharmaceutically acceptable carriers and/or additives
WO2016029328A1 (fr) * 2014-08-25 2016-03-03 Muciderm S.A Composition pharmaceutique pour la prévention, le traitement et la suppression du psoriasis, contenant de la bave d'escargot, de la camomille et du miel
US10350247B2 (en) 2014-08-25 2019-07-16 Muciderm S.A. Pharmaceutical composition for preventing, treating, and curing psoriasis including snail slime, chamomile, and honey
WO2016054757A1 (fr) * 2014-10-10 2016-04-14 Muciderm S.A. Composition pharmaceutique pour la prévention, le traitement et la suppression de la rosacee, contenant de la bave d'escargot, de la camomille et du miel
US20170281690A1 (en) * 2014-10-10 2017-10-05 Muciderm S.A. Pharmaceutical composition for preventing, treating and curing rosacea, comprising snail slime, chamomile and propolis
US11045503B2 (en) 2014-10-10 2021-06-29 Muciderm S.A. Pharmaceutical composition for preventing, treating and curing rosacea, comprising snail slime, chamomile and propolis
EP3124030A1 (fr) * 2015-07-30 2017-02-01 Aristotle University Of Thessaloniki-Research Committee Procédé de production d'un extrait d'escargot petit-gris (cornu aspersum) et ledit extrait
IT201700122874A1 (it) * 2017-10-31 2019-05-01 Roberto Leone Composizione edibile a base di bava di lumaca per la conservazione degli alimenti e per la sanificazione di superfici, ambienti e strumenti
CN110314247A (zh) * 2019-08-02 2019-10-11 深圳国佳产业基金管理有限公司 蜗牛精华提取物活性敷料及其制备方法
WO2022150018A1 (fr) * 2021-01-08 2022-07-14 Aden International Co.,Ltd. Nouveau procédé d'isolement, de purification et de caractérisation de substances semblables à l'héparine provenant de mucus d'escargot (achatina fulica) et utilisations associées

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