WO1995023604A1 - Compositions de stimulation de la croissance cellulaire renfermant de l'aloesine - Google Patents

Compositions de stimulation de la croissance cellulaire renfermant de l'aloesine Download PDF

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Publication number
WO1995023604A1
WO1995023604A1 PCT/KR1995/000016 KR9500016W WO9523604A1 WO 1995023604 A1 WO1995023604 A1 WO 1995023604A1 KR 9500016 W KR9500016 W KR 9500016W WO 9523604 A1 WO9523604 A1 WO 9523604A1
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Prior art keywords
aloesin
cell growth
growth stimulating
cells
composition
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PCT/KR1995/000016
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English (en)
Inventor
Kyeong Ho Kim
Jeong Hill Park
Chung Ki Sung
Jin Woong Kim
Jae Sue Choi
Seung Ki Lee
Yeong Shik Kim
Byeong Hwa Son
Myung Hee Chung
Young In Park
Kyu Won Kim
Chong Kil Lee
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Namyang Aloe Co., Ltd.
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Application filed by Namyang Aloe Co., Ltd. filed Critical Namyang Aloe Co., Ltd.
Priority to AU19613/95A priority Critical patent/AU1961395A/en
Publication of WO1995023604A1 publication Critical patent/WO1995023604A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; 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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60—Sugars; Derivatives thereof
    • A61K8/602—Glycosides, e.g. rutin
    • 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

  • the present invention relates to a cell growth stimu ⁇ lating composition containing aloesin as an active ingre ⁇ host. More specifically, the present invention relates to a composition for stimulating cell growth, especially for stimulating epidermal cells and hepatic cells, which contains aloesin as an active ingredient.
  • Aloe is a medicinal herb belonging to Liliaceae which has been widely and popularly used from ancient times.
  • the record of the first use of aloe as a medicine can be found from a mural painting in Egyptian temple in which Aloe vera is described as the plant of immortality.
  • the specific record of the use of aloe can be found in Sumeri- an clay tablets around 1750 B.C.
  • Egyptian Book of Remedies around 1550 B.C. describes the use of aloe as an agent for treatment of infections and for skin massage and also the method for preparing aloe as a medi ⁇ cine. Meanwhile, in A.D. 74 Dr.
  • Aloe vera is a very effective laxative and has the effect of treating wound, furuncle, chilblained skin, etc., and also alleviating genital ulcers and tonsillitis.
  • various aloe plants for example, Aloe perryi, Aloe forex, Aloe barbadenis, etc., have been popularly used for a long time in the form of suntan ointment, suntan powder, cosmetic cream, lotion, shampoo, and the like.
  • aloe has various pharmacological activities including wound healing effect, anti-inflammatory activity, anti-ulcer activity, anti- cancer activity ( cell differentiation activity) , anti ⁇ microbial activity, immunomodulatory activity, cell growth stimulating activity, skin penetrating activity and the like .
  • wound healing effect anti-inflammatory activity
  • anti-ulcer activity anti- cancer activity
  • anti ⁇ microbial activity anti ⁇ microbial activity
  • immunomodulatory activity cell growth stimulating activity
  • skin penetrating activity and the like .
  • no systematic and definite result of study regarding that each pharmacological activity of aloe plants is derived from what kind of the components has been reported yet.
  • aloesin one of the effec ⁇ tive components of aloe plants , has an excellent cell growth stimulating activity , and then completed the present invention.
  • aloesin as an agent for stimulating cell growth, especially growth of epidermal cells and hepatic cells.
  • Figure 1 is the result of Western Blot Analysis which determines the influence of aloesin on the activity of antibody cdk2 in hepatoma cell line sk-Hep-l according to Example 7-2;
  • Figure 2 is the result of Western Blot Analysis which determines the influence of aloesin on the activity of antibody cyclin E in hepatoma cell line sk-Hep-l according to Example 7-2;
  • Figure 3 is the result of Western Blot Analysis which determines the influence of aloesin on the activity of antibody cdc25 in hepatoma cell line sk-Hep-l according to Example 7-2;
  • Figure 4 is the result of Immune Complex Kinase Assay which determines the influence of aloesin on the activity of antibody cdk2 in hepatoma cell line sk-Hep-l according to Example 7-3.
  • a cell growth stimulating composition containing aloesin, which is one of the effective components of aloe plants, as an active ingredient is provided.
  • the composition of the present invention can effectively stimulate the growth of cells, especially epidermal cells and hepatic cells.
  • aloe has been used generally in the form of a dry extract which is obtained by extracting leaves of various aloe plants belonging to Aloe genus such as Aloe vera with water and then evaporating the resulting aqueous extract to dryness.
  • the dry extract of aloe still contains the other effective components of aloe, in addition to certain components having the desired pharmacological effect, it may occasionally cause the undesirable side effects.
  • the effective component of aloe i.e. aloesin can be utilized without any side effects other than the desired effect.
  • Aloesin isolated and utilized in the present invention is a component extacted from leaves of Aloe vera and is represented by the following structure [molecular formula: C 19 H 22°9' molecular weight: 394] :
  • aloesin has an effect for blocking ultraviolet ray from skin [see Strickland, F.M. , Pelley, R.P., Hill, D. and Kripke, M.L., J. Investigative Dermatology, 98: 653, 1992].
  • aloesin has an activity as mushroom tyrosinase inhibitor [see Akira Yagi, Toshimitsu Kanbara and Naoka Morinobu, Planta Medica, 515- 517, 1987].
  • any pharmacological use of aloesin as a medicinal agent has not been reported yet.
  • aloesin which is obtained by treating aloe plants according to known method and then isolating and purifying aloesin has a potent effect for stimulating cell growth, especially for stimulating growth of skin epithe- lial cells and hepatic cells.
  • aloesin which can be administered to the patients suffer ⁇ ing from skin wound
  • aloesin can be administered gener ⁇ ally in an amount of O.lmg to 500mg, preferably in an amount of 0.4mg to 125mg, per a day, per kg of body weight of adult man to obtain the desired effect.
  • a suitable daily dosage of aloesin can be varied depending on the severity of diseases and is generally O.lmg to 500mg, preferably 0.4mg to 125mg, per kg of body weight of adult man to provide the desired effect for stimulating the growth of hepatic cells.
  • Aloesin which is used in the present invention can be obtained by treating aloe plants according to the method known in this technical field, for example, the method disclosed by Yagi, et al. [see Akira Yagi, Kenji Makino and Itsuo Nishioka, Chem. Pharm. Bull. 25(7), 1771-1776, 1977].
  • aloesin can be obtained by cleanly washing fresh aloe plants, for example, whole leaves of Aloe vera, with water, extracting the washed plant with a lower alcohol such as methanol, ethanol, propanol, buta- nol, etc., a lower ketone such as acetone, methylethyl ketone, etc., or a mixture thereof, suspending the result ⁇ ing extract in water, extracting again the suspension with a lower ether such as diethyl ether, etc., a lower ester such as ethyl acetate, etc.
  • a lower alcohol such as methanol, ethanol, propanol, buta- nol, etc.
  • a lower ketone such as acetone, methylethyl ketone, etc.
  • a mixture thereof suspending the result ⁇ ing extract in water, extracting again the suspension with a lower ether such as diethyl ether, etc., a lower ester such as e
  • a halogenated hydrocarbon such as carbon tetrachloride, dichloromethane, chloroform, etc., or a mixture thereof, extracting again the aqueous layer with a lower alcohol, and then subjecting the alco ⁇ hol extract to silica gel column chromatography eluting with a mixed solvent consisting of halogenated hydrocarbon and lower alkanol in the mixing ratio of 2:1 to 100:1, preferably 5:1 to 20:1, or a mixed solvent consisting of lower ester, lower alcohol and water in the mixing ratio of 20:1:0 to 6:4:2 to isolate aloesin in a pure state.
  • a mixed solvent consisting of halogenated hydrocarbon and lower alkanol in the mixing ratio of 2:1 to 100:1, preferably 5:1 to 20:1, or a mixed solvent consisting of lower ester, lower alcohol and water in the mixing ratio of 20:1:0 to 6:4:2 to isolate aloesin in a pure state.
  • aloesin When aloesin is clinically used, aloesin can be admin ⁇ istered to patients suffering from burn, trauma, etc, who require cell growth stimulating effect, or patients who has a damage of tissue cells caused by hepatoma, hepati ⁇ tis, etc. , in the form of conventional pharmaceutical preparations according to a conventional manner utilized in the field of pharmaceutical preparation.
  • the pre- ferred pharmaceutical preparations which can be suitably used for this purpose include preparations for oral admin ⁇ istration such as tablets, hard or soft capsules, solu ⁇ tions, suspensions, etc., injections for parenteral admin ⁇ istration such as injectable solutions or suspensions, etc., preparations for topical use such as ointments, creams, gels, lotions, etc., and the like.
  • Such pharma ⁇ ceutical preparations can also be prepared by utilizing pharmaceutically acceptable carriers conventionally used in this technical field, for example, excipients, binders, disintegrants, lubricants, solubilizing agents, suspending agents, preservatives or extending agents for preparations for oral administration, stabilizers, preservatives, solu ⁇ bilizing aids, buffers, agents for isotonicity, etc., for injections, or aqueous or oily ointment bases, anti- oxidants, preservatives, extending agents, etc., for topical preparations.
  • pharmaceutically acceptable carriers for example, excipients, binders, disintegrants, lubricants, solubilizing agents, suspending agents, preservatives or extending agents for preparations for oral administration, stabilizers, preservatives, solu ⁇ bilizing aids, buffers, agents for isotonicity, etc., for injections, or aqueous or oily ointment bases, anti
  • the preparation for stimulating growth of epidermal cells and hepatic cells which contains aloesin according to the present invention can be prepared so that a unit dosage form can contain the above-mentioned preferred daily dosage of aloesin or its 1/2, 1/3, 1/4, 1/5 or 1/6. This unit dosage form can be administered 1 to 6 times per day.
  • aloesin for stimulating cell growth can be observed according to the determination of cell count using a he ocytometer or Coulter counter as described by Doyle, et al. [see Doyle, A., Griffiths, J.B. and Newell, D.G., Cell & Tissue Culture: Laboratory Procedures, 10E, WILEY (1993)], the quantitative method by means of a dye such as Hoechst 38825, acridine orange, diaminobenzoic acid (DABA) , a method for determination of DNA synthetic rate using a radioisotope [ 3 H]-methyl-thymidine, and recently a method for determination of cell growth without using any radioisotope, for example, methods for dete ⁇ ai- nation of a cell component BCECF (2 ⁇ ,7 '-bis-(carboxy- ethyl)-5(or 6)-carboxyfluoresin acetoxymethyl ester) [see Kolber et al., Journal of Im un
  • a method for determination of certain components including BCECF, neutral red, 4-methylumbellif- erone phosphate, etc. , takes advantage of characteristic features of these components that they contain a uniform amount of enzymes or shows an increased activity in pro ⁇ portion to the increase of cell counts.
  • these methods have some disadvantages in that their sensitivity is lower than that of methods for determination of induc ⁇ tion rate of a radioisotope [ 3 H] -methyl-thymidine.
  • the following examples of the present inven- tion were conducted utilizing a method for determination of induction rate of [ H]-methyl-thymidine and a MTT assay for determination of activity of cellular reductase to determine the effect of aloesin for stimulating cell growth.
  • Western Blot Analysis and Immune Complex Kinase Assay were conducted to identify that the cell growth stimulating activity of aloesin is derived from the stimulation of DNA synthesis in cells without any cell damage.
  • hepatoma cell line sk-Hep-l skin epithelial cell line HT-1080, kidney epithelial cell line COS-7 and gastric cancer cell line KATO-111 were obtained from Cancer Institute of Seoul National University in Korea, and then incubated with a culture solution, which is prepared by dissolving DMEM (Dulbecco's Modified Ea ⁇ gle's Medium, made by Gibco) in 1 liter of deionized water, adjusting the resulting solution to pH 7.4, adding 10% calf fetal serum, 1X10 "7 M insulin and 50mg/l of gentamycine thereto, and then sterilizing the resulting mixture with a Millipore filter, in an incubator of 37°C while maintaining 5% carbon dioxide.
  • DMEM Dissolving DMEM (Dulbecco's Modified Ea ⁇ gle's Medium, made by Gibco) in 1 liter of deionized water, adjusting the resulting solution to pH 7.4, adding 10% calf fetal serum,
  • the extracted butanol fractions were combined and then subjected to column chromatography on silica gel eluting with a mixed solvent of ethyl acetate/ ethanol/water (10:1:0.5) or a mixed solvent of chloroform/methanol (5:1) to obtain the desired aloesin- containing fraction.
  • the obtained aloesin-containing fraction was subjected again to silica gel column chroma ⁇ tography eluting with a mixed solvent of ethyl acetate/methanol/water (10:1:0.2) or a mixed solvent of chloroform/methanol (7:1) to obtain more purified aloesin- containing fraction.
  • This aloesin-containing fraction was recrystallized from ethyl acetate or a mixture of ether and methanol to obtain about 70mg of the desired aloesin.
  • Aloesin obtained above shows NMR and mass spectrum data which are identical to those disclosed in the refer ⁇ ence [see John M. Conner, Alexander I. Gray, Tom Reynolds and Peter G. Waterman, Phytochemistry , 29, 941-944, 1990]. specifically, when about 3mg of the obtained aloesin is dissolved in 600 ⁇ l of DMSO-d g (99.8%) to measure its NMR spectrum, NMR spectrum of aloesin shows characteristic peaks at 2.20, 2.63, 3.75, 6.07, 6.65 pp . In addition, mass spectrum (EI+, 70eV) of aloesin shows peaks of 394 (molecular ion peak) , 245 (basic peak) , 304, 203, 163, etc.
  • EXAMPLE 2 Effect of aloesin for stimulating cell growth on human hepatoma cells
  • DMEM Dulbecco's Modified Eagle's Medium, made by Gibco
  • DMEM Dulbecco's Modified Eagle's Medium
  • pH 7 . 4 10% calf fetal serum, 1X10 "7 M insulin and 50mg/l of gentamycin were added thereto .
  • the mixture was sterilized through a
  • the treated sample was treated w ith 3 H- label led thymidine in the concentrati on o f l ⁇ Ci /ml .
  • the medium was removed from each well , and the cells were fixed with methanol and then washed with PBS .
  • the cells were washed two times with 10 % trichloroacetate to remove unreacted radioactive thymidine , dissolved in IN sodium hydroxide and then neutralized with IN hydrochloric acid.
  • the radio- activity introduced into DNA was measured with a scintil ⁇ lation counter (Pharmacia 1024 ) .
  • the measured result is described in the following Table 1.
  • EXAMPLE 3 Effect of aloesin for stimulating cell growth on human epithelial cells
  • DMEM Dulbecco's Modified Eagle's Medium, made by Gibco
  • DMEM Dulbecco's Modified Eagle's Medium
  • pH 7.4 10% calf fetal serum
  • 1X10 M insulin and 50mg/l of genta ycin were added thereto.
  • the mixture was sterilized through a Millipore filter to obtain the culture solution.
  • Human epithelial cell line HT-1080 which was obtained from Cancer Institute of Seoul National University was inoculated into the culture solution in the ratio of 1X10 6 cells per 25cm 2 of the surface area of T flask and then incubated for 48 hours in an incubator of 37°C while maintaining 5% carbon dioxide.
  • the cultures were transferred to a 24-well plate, subcultured for one day and then treated with 100 to l ⁇ g/ml of aloesin.
  • the cul ⁇ tures were treated with the same amount of PBS (phosphate buffered saline) instead of aloesin.
  • the treated sample was treated with 3 H- labelled thymidine in the concentration of l ⁇ Ci/ml. After 12 hours, the medium was removed from each well, and the cells were fixed with methanol and then washed with PBS.
  • the cells were washed two times with 10% trichloro- acetate to remove unreacted radioactive thymidine, dis ⁇ solved in IN sodium hydroxide and then neutralized with IN hydrochloric acid. Then, the radioactivity introduced into DNA was measured with a scintillation counter (Phar ⁇ macia 1024) . The measured result is described in the following Table 2.
  • DMEM Dulbecco's Modified Eagle's Medium, made by Gibco
  • DMEM Dulbecco's Modified Eagle's Medium
  • pH 7.4 10% calf fetal serum
  • 1X10 ⁇ 7 M insulin and 50mg/l of gentamycin were added thereto.
  • the mixture was sterilized through a Millipore filter to obtain the culture solution.
  • Mon ⁇ key's kidney cell line C0S7 which was obtained from Cancer Institute of Seoul National University was inoculated into the culture solution in the ratio of 1X10 6 cells per 25cm 2 of the surface area of T flask and then incubated for 43 hours in an incubator of 7°C while maintaining 5% carbon dioxide.
  • the cultures were transferred to a 24-well plate, subcultured for one day and then treated with 100 to O.OOOl ⁇ g/ml of aloesin.
  • the cultures were treated with the same amount of PBS (phos ⁇ phate buffered saline) instead of aloesin.
  • PBS phos ⁇ phate buffered saline
  • the treated sample was treated with 3 H-labelled thymidine in the concentration of l ⁇ Ci/ml.
  • the medium was removed from each well, and the cells were fixed with methanol and then washed with PBS.
  • the cells were washed two times with 10% trichloroacetate to remove unreacted radioactive thymidine, dissolved in IN sodium hydroxide and then neutralized with IN hydrochloric acid. Then, the radio- activity introduced into DNA was measured with a scintil ⁇ lation counter (Pharmacia 1024) . The measured result is described in the following Table 3.
  • EXAMPLE 5 Effect of aloesin for stimulating cell growth in primary cultures of rat hepatic cells
  • hepatic cells were conducted accord ⁇ ing to the method of Dickens and Peterson [see Dickens and Peterson, Biochem. Pharmacol. 29, 1231, 1980].
  • rat was anesthetized with intraperi- toneal injection of urethane in an amount of lg/kg.
  • the abdomen was shaved to remove hairs and then the shaved portion was disinfected with iodine tincture.
  • the subse- guent experimental procedures were carried out under sterilized condition.
  • the abdomen was incised along the center line and then both ends were cut down to expose the hepatic portal to which a 18 gauge catheter was connected.
  • Inferior vena cava was cut down, and then Hank's balance salt solution was infused in the flow rate of 15 to 20ml per minute through the catheter to remove the blood in liver.
  • the inferior vena cava was ligated, the chest was opened to expose the superior vena cava to which a 16 gauge catheter was connected. The fluid flowing out through the superior vena cava was recycled. This procedure should be practiced while maintaining the temperature of 37°C and under continuously injection of a gas consisting of 95% oxygen/5% carbon dioxide at the rate sufficient to saturate the perfusion.
  • Collagense (Sigma, C1030) was added to the perfusion in the concentration of 0.05% to 0.06% and this mixture was perfused for 15 minutes during which the swelling of liver C an be visually observed.
  • the liver was completely extracted, transferred into 60ml of the perfusion and then cut down with a sterilized scissor.
  • the separated cells were filtered with a 250 ⁇ m nylon membrane.
  • the filtrate was centrifuged at 50Xg for 40 minutes and washed three times with the perfusion.
  • the final precipitate was dispersed in the cell culture solution.
  • the survival rate, of the prepared liver cell dispersion was determined by adding 900 ⁇ l of the staining solution consisting of 0.4% tryphan blue in physiological saline to lOO ⁇ l of the dispersion, standing this mixture for 5 minutes and then counting the stained died cells and the unstained living cells under a microscope and calcu ⁇ lating their ratio. Then, only the liver cell dispersion having a survival rate of 90% or more was selected, to which the experiment was continuously practiced.
  • the sample was treated with 3 H-labelled thymi ⁇ dine in the concentration of l ⁇ Ci/ml, and then, after 12 hours, the culture medium was removed from each well.
  • the cells were fixed with methaol and washed with PBS and then washed two times with 10% trichloroacetate to remove unreacted radioactive thymidine.
  • the cells were dis ⁇ solved in IN sodium hydroxide and neutralized with IN hydrochloric acid, and then the radioactivity introduced into the cells was measured by means of a scintillation counter (Pharmacia 1024) . The result thereof is de ⁇ scribed in the following Table 4. Table 4. Amount of radioactive thymidine introduced into DNA of primary cultured hepatic cells depending on aloesin concentration
  • EXAMPLE 6 Effect of aloesin for stimulating cell growth on human hepatoma cells (MTT test.
  • DMEM Dulbecco's Modified Eagle's Medium, made by Gibco
  • DMEM Dulbecco's Modified Eagle's Medium
  • 1X10 "7 M insulin and 50mg/l of gentamycin were added thereto.
  • the mixture was sterilized through a Millipore filter to obtain the culture solution.
  • Human hepatoma cell line sk-Hep-l which was obtained from Cancer
  • the following experiment was conducted to identify that the cell growth stimulating activity of aloesin is derived from the stimulation of DNA synthesis in cells without any cell damage.
  • Hepatoma cell line sk-Hep-l which was obtained from Cancer Institute of Seoul Ntional University was spread over a 10mm culture dish in the concentration of 8-10xl0 5 cells per dish and then incubated in DMEM (Dulbecco's Modified Eagle's Medium, made by Gibco) for 24 hours. Cells were treated with 4mM hydroxyurea for 14 hours to synchronize the cells. Then, the medium in the culture dish was replaced with aloesin-DMEM medium and the cells were incubated again for 24 hours.
  • DMEM Dulbecco's Modified Eagle's Medium, made by Gibco
  • the cells were washed two times with cold PBS, harvested using lOO ⁇ l, per dish, of RIFA buffer (1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15% NaCl, ImM phenylmethyl-sulfonylfluoride, 0.2mM EDTA, 50mM NaF, 0.2mM Na 3 V0 4 , 30mM ⁇ -glycerophosphate, 100 kallikrein inactivat ⁇ ing units [KIU] aprotinin) , and then centrifuged with 12000rpm to separate the supernatant. The separated supernatant was used as the cell lysate which is the test protein in the following Western Blot Analysis and Immune
  • the protein-blotted PVDF membrane was completely dried at room temperature, incubated in 5% nonfat dry milk solution for one hour and then washed three times with TBST solution (50mM Tris, pH 7.4, 150mM NaCl, 0.05% Tween 20) for 5 minutes each time, and primary antibodies cdk2 , cyclin E and cdc25 which were diluted with TBST solution at the ratio of 1:2000, 1:5000 and 1:5000, respectively, were added thereto.
  • TBST solution 50mM Tris, pH 7.4, 150mM NaCl, 0.05% Tween 20
  • the PVDF membrane was incubated for one hour, washed three times with TBST solution for 5 minutes each time, and then incubated with peroxidase-linked secondary antibody (anti- mouse Ig) solution diluted with TBST solution at the ratio of 1:15000 for one hour.
  • the membrane was washed again four times with TBST solution for 5 minutes each time, and then reacted with a mixed solution of the same amounts of reagent 1 and reagent 2 of ECL (Enhance Chemilumines- cence) immunoblotting kit (made by Amersham) .
  • ECL End Chemilumines- cence
  • lOO ⁇ g of the cell lysate isolated from aloesin-treated hepatoma cell sk-Hep-l obtained in the above 1 was pre- purified with BSA precoated-Sepharose, and the supernatant was separated and then incubated with cdk2 antibody (5 ⁇ l) .
  • Immuno-precipitate was obtained by adding the protein A bead and washed three times with RIFA buffer and then three times with two-fold HI kinase assay buffer (lOOmM Tris,pH 7.6, 20mM MgCl 2 , 2mM DTT, l ⁇ g/ml antipain, leupep- tin, pepstatin) .
  • HI kinase assay of the immuno- precipitate was conducted by reacting the immuno- precipitate with buffer solution (50 ⁇ M ATP, lO ⁇ Ci [r- 32 P]ATP) containing 0.6mg/ml of histone HI and, after 12% SDS-PAGE, measuring the phosphorylation of histone HI by means of autoradiography.
  • buffer solution 50 ⁇ M ATP, lO ⁇ Ci [r- 32 P]ATP
  • aloesin increases the activities of cdk2, cyclin E and cdc25 to stimulate the synthesis of DNA, and ulti ⁇ mately exhibits an excellent cell growth stimulating activity.
  • Composition 1 (Soft capsule)
  • the soft capsules containing 5mg of aloesin per cap ⁇ sule were prepared using the following components accord ⁇ ing to the method for capsule preparation described in Korea Pharmacopeia. Component Content (mg/cap.)
  • Composition 2 (Tablet)
  • the tablet preparations containing 5mg of aloesin per tablet were prepared using the following components ac ⁇ cording to the method for tablet preparation described in Korea Pharmacopeia.
  • Composition 3 (Injection)
  • the injections containing 2mg of aloesin per ampoule were prepared using the following components according to the method for injection preparation described in Korea Pharmacopeia. Component Content (mg/amp . )
  • composition 4 (Ointment)
  • ointment preparations containing 5mg of aloesin per lg were prepared using the following components ac ⁇ cording to the method for ointment preparation described in Korea Pharmacopeia.

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Abstract

Composition de stimulation de la croissance cellulaire renfermant de l'aloésine à titre d'ingrédient actif. Plus spécialement, il s'agit d'une composition de stimulation de la croissance cellulaire, et notamment de stimulation des cellules hépatiques et épidermiques, renfermant de l'aloésine à titre d'ingrédient actif. L'aloésine est un constituant de l'aloès vrai dont un effet de protection de la peau contre le rayonnement ultraviolet, ainsi qu'une activité d'inhibition de la tyrosinase des champignons, sont déjà connus de l'état de la technique. Cependant, on a mis en évidence que l'aloésine a une nouvelle utilisation pharmacologique en tant qu'agent de stimulation de la croissance cellulaire qui ne fait l'objet d'aucune divulgation dans l'état de la technique.
PCT/KR1995/000016 1994-03-03 1995-03-02 Compositions de stimulation de la croissance cellulaire renfermant de l'aloesine WO1995023604A1 (fr)

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AU19613/95A AU1961395A (en) 1994-03-03 1995-03-02 Cell growth stimulating compositions containing aloesin

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004024899A1 (fr) * 2002-09-13 2004-03-25 Medi-Cult A/S Facteur de croissance pour culture cellulaire
EP1503777A4 (fr) * 2002-05-03 2008-02-20 Unigen Pharmaceuticals Inc 7-hydroxy chromones utilises en tant que puissants antioxydants
CN102584807A (zh) * 2012-02-06 2012-07-18 上海华震科技有限公司 一种芦荟苦素的乙醇浸提工艺
US8852657B2 (en) 2007-01-09 2014-10-07 Unigen, Inc. Chromones as therapeutic agents
US20170014461A1 (en) * 2015-07-15 2017-01-19 Unigen, Inc. Compositions, Methods, and Medical Compositions for Treatment of and Maintaining the Health of the Liver
CN109498538A (zh) * 2018-12-28 2019-03-22 广州市白云区研美化妆品厂 具有保湿美白功效的组合物、化妆品、精华液及其制备方法

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EP1503777A4 (fr) * 2002-05-03 2008-02-20 Unigen Pharmaceuticals Inc 7-hydroxy chromones utilises en tant que puissants antioxydants
US7678772B2 (en) 2002-05-03 2010-03-16 Unigen Pharmaceuticals, Inc. 7-hydroxy chromones as potent antioxidants
US8110555B2 (en) 2002-05-03 2012-02-07 Unigen, Inc. 7-hydroxy chromones as potent antioxidants
US9078891B2 (en) 2002-05-03 2015-07-14 Unigen, Inc. 7-hydroxy chromones as potent antioxidants
WO2004024899A1 (fr) * 2002-09-13 2004-03-25 Medi-Cult A/S Facteur de croissance pour culture cellulaire
US8852657B2 (en) 2007-01-09 2014-10-07 Unigen, Inc. Chromones as therapeutic agents
CN102584807A (zh) * 2012-02-06 2012-07-18 上海华震科技有限公司 一种芦荟苦素的乙醇浸提工艺
US20170014461A1 (en) * 2015-07-15 2017-01-19 Unigen, Inc. Compositions, Methods, and Medical Compositions for Treatment of and Maintaining the Health of the Liver
RU2755478C2 (ru) * 2015-07-15 2021-09-16 Юниджен, Инк. Композиции, способы и медицинские композиции для лечения и поддержания здоровья печени
US11331359B2 (en) * 2015-07-15 2022-05-17 Unigen, Inc. Compositions, methods, and medical compositions for treatment of and maintaining the health of the liver
US20220265747A1 (en) * 2015-07-15 2022-08-25 Unigen, Inc. Compositions, Methods, and Medical Compositions for Treatment of and Maintaining the Health of the Liver
CN109498538A (zh) * 2018-12-28 2019-03-22 广州市白云区研美化妆品厂 具有保湿美白功效的组合物、化妆品、精华液及其制备方法

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