WO2000061162A1 - Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa - Google Patents

Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa Download PDF

Info

Publication number
WO2000061162A1
WO2000061162A1 PCT/US2000/008711 US0008711W WO0061162A1 WO 2000061162 A1 WO2000061162 A1 WO 2000061162A1 US 0008711 W US0008711 W US 0008711W WO 0061162 A1 WO0061162 A1 WO 0061162A1
Authority
WO
WIPO (PCT)
Prior art keywords
mixture
curcumin
tetrahydrocurcuminoids
curcuminoids
linking
Prior art date
Application number
PCT/US2000/008711
Other languages
French (fr)
Other versions
WO2000061162A9 (en
Inventor
Muhammed Majeed
Vladimir Badmaev
Original Assignee
Sabinsa Corporation
Sami Chemicals And Extracts (P) Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to US12854099P priority Critical
Priority to US60/128,540 priority
Application filed by Sabinsa Corporation, Sami Chemicals And Extracts (P) Ltd. filed Critical Sabinsa Corporation
Publication of WO2000061162A1 publication Critical patent/WO2000061162A1/en
Priority claimed from US09/972,150 external-priority patent/US6653327B2/en
Publication of WO2000061162A9 publication Critical patent/WO2000061162A9/en

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q17/00Barrier 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
    • A61Q17/04Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/12Ketones
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K41/00Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/30Cosmetics or similar toilet preparations characterised by the composition containing organic compounds
    • A61K8/33Cosmetics or similar toilet preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/35Ketones, e.g. benzophenone
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/02Preparations for care of the skin for chemically bleaching or whitening the skin

Abstract

The present invention is directed to a mixture of tetrahydrocurcuminoids which can be used for regulating random intracellular protein crosslinking, optimizing cell electric potential, treating a premalignant state or a malignancy, increasing intracellular transglutaminase activity, and reducing 'age spots' due to lipofuscin formation.

Description

USE OF TETRAHYDROCURCUMINOIDS TO REGULATE PHYSIOLOGICAL AND PATHOLOGICAL EVENTS IN THE SKIN AND MUCOSA
Background of the invention
A body cell owes its functioning in large part to its cytoskeleton, the structural
framework of the cell. The cytoskeleton is formed by components in the cytoplasm
that maintain cell shape, stabilize cell attachments to neighboring cells, and facilitate
specialized cell functions like endocytosis, exocytosis and cell motility. The
cytoskeleton includes several filamentous structures like microtubules,
microfilaments, intermediate filaments, and microtrabecular lattice. Proteins
participating in the formation of a cytoskeleton, especially microtubular-associated
proteins (MAPS) after being manufactured by the cell, are further adapted to the
cytoskeleton function in a process called post-translational modification, or cross-
linking. This process of proteins cross-linking is essential to stabilizing the
cytoskeleton, the event necessary to maintain cell homeostasis, physiological
processes, and cell development and maturation.
The post-translational modification of proteins involves several chemical
reactions. Protein phosphorylation exemplifies one of the most common
mechanisms of post translation modification of cellular proteins. Other examples
include methylestrification of proteins, ADP-ribosylation, protein glycosylation,
histone acetylation, hypusine formation and transglutaminase-catalyzed reactions.
The electric potential of a cell is an aspect of cellular physiology important to
the understanding of the mechanism of the invention. The electric potential of a cell is maintained by a complex balance between negatively and positively charged
molecules which results in a physiologic resting potential inside the cell of
approximately - 70 mV. This value is indicative of the cell's well-being. On the other
hand, the inability of a cell to maintain an optimal resting potential occurs in
pathological conditions, e.g. physical or chemical assaults, a disease process, and
the wear and tear due to aging. Optimal resting potential is conductive to the
physiological processes including post-translational protein modification and cross-
linking. It also holds true that the intact physiological cross-linking of proteins is
indicative of a cell's ability to maintain optimal resting electric potential,
Positive effects of cross-linking
Cross-linking of proteins, as it has been previously mentioned, is an outcome
of the post translational modification of proteins. Some of the most common cross¬
links identified in proteins forming cytoskeleton are Neee-(ggg-glutamyl)lysine and
N,N-bis(gggglutarnyl)polyamine. The formation of these two types of cross-links is
facilitated by the catalytic activity of a group of enzymes known as
transglutaminases. One of the well known transglutaminases in the body is
coagulation protein Factor XIII. This Factor causes cross-linking of the fibrin, which
acts as a stabilizing agent on blood clot formation. This type of modification is an
example of a life saving mechanism-based on protein cross-linking, that prevent
exsanguination in a cut or a wound.
Transglutaminases are responsible for facilitating a physiological process of
protein cross-linking, which under normal conditions leads to cell differentiation, thus the cell is able to fulfill its purpose, e.g. mucus secreting cell, phagocytic cell,
epithelial and tegumen cell protecting the tissue structure. Transglutaminase
activity, or its absence, plays a critical role in the differentiation of cancer cells,
especially skin cancer melanoma. Melanoma is a rapidly spreading cancer
consisting of undifferentiated skin cells, and the degree of poor cell differentiation
correlates well with the suppressed activity of transglutaminase. On the other hand,
experimental induction of transglutaminase activity led to promising results in
slowing-down or reversing melanoma progression. Examples of transglutaminase
inducers are retinoic acid and methylxantines. It appears that the therapeutic
potential of retinoids in melanoma is related to their ability to activate intracellular
transglutaminases.
Detrimental cross-linking
Cross-linking of intracellular proteins is, however, not always a positive event.
When a random and massive cross-linking is precipitated by an abrupt, non-
physiological stimulus, an accelerated aging of the cell occurs. Due to the nature of
random crosslinking, which defies the purpose of cell organization and
differentiation, this may lead not only to premature cell aging and death, but also can
lead to a pathology including malignancy. For example, the UVA-generated free
radicals in skin cells can cause premature cross-linking of proteins and liberation of
proteolytic enzymes. This latter event leads to the destruction of the cytoskeleton,
and the rapid deterioration of cell organization which compromises cell functioning.
It is of special interest to mention here the term lipofuscin, nicknamed "the aging pigment" or "liver spots", which occurs in the epidermis and mucous
membranes probably as a result of disrupted cellular and systemic metabolism.
Lipofuscin has been shown to result from the oxidative degeneration of mitochondria
and/or lysosomes in the cells. Lipofuscin is postulated to be a result of a metabolic
error which results in hypoanabolism combined with hyperkatabolism. It is
postulated here that lipofuscin formation is in part a manifestation of the disrupted
process of post-translational protein modification, and that this process intensifies
with any systemic disease process, including the aging process.
Related art
As previously mentioned, transglutaminases are responsible for facilitating
the physiological process of protein cross-linking, which under normal conditions
leads to cell differentiation, maturation and timely aging. There is considerable
evidence that intracellular transglutaminase activity may be inhibited in certain
pathology, for example malignancy, e.g. melanoma. It appears that therapeutic
potential of transglutaminase inducers like retinoids and methylxantines in
melanoma is related to their ability to activate intracellular transglutaminases. This
in turn would lead to differentiation of cells, which in clinical practice would lead to a
slow down of the disease process due to a decrease in the number of
undifferentiated malignant cells.
Curcumin derivatives including tetrahydrocurcumin and curcuminoids are well
recognized in literature as a group of phenolic antioxidants (Majeed et al.
Curcuminoid book). Majeed et al., U.S. Patent No. 5,861 ,415 established that the combination of naturally occurring curcuminoids isolated from turmeric root, i.e.
curcumin, demethoxy curcumin and bisdemethoxy curcumin produce a broad range
of antioxidant protection described as bioprotectant action. Bioprotectant action,
characterized by the prevention of free radical formation and intervention in
scavenging free radicals, is exerted by tetrahydrocurcumin(oids) (THC) as well as
curcumin(oids) (Majeed et a\.,Tumeric and the Healing Curcuminoids, Keats
Publications 1996 and Majeed et al., Curcuminoids Antioxidant Phytonutrients,
NutriScience Publishers 1995). However, tetrahydrocurcumin(oids) show superior
free radical scavenging properties, as compared to curcumin or curcuminoids
(Majeed et al. Curcuminoid book). Another advantage of tetrahydrocurcuminoids is
that they are not as highly colored curcuminoids and thus can be applied topically
without significant staining.
Recently curcumin but not tetrahydrocurcumin was found to have a
preventive effect on cross-linking of collagen in diabetic rats. (Sajithlal GB, Chithra
P, Chandrakasan G., Biochem Pharmacol 1998 Dec 15;56(12):1607-14). However,
the anti cross-linking effect of curcumin was attained with a very high oral dose of
the compound (200 mg/kg body wt.). It was also noted that the preventive effect of
curcumin on the advanced cross-linking of collagen was more pronounced than its
therapeutic effect. The authors of this study explained that the anti cross-linking
mechanism of curcumin is due to quenching the free radicals, which in turn would
prevent the accelerated cross-linking of collagen in diabetes. This study supports the
use of curcumin to prevent and to some degree treat excessive cross-linking of
proteins in conditions characterized by accelerated cross-linking such as diabetes. Description of the invention
The mechanism of the invention is in preserving homeostasis and functioning
of cells based on its regulatory, both inhibitory and stimulating, effect on the cross-
linking of intracellular proteins. This regulatory mechanism, previously unknown for
curcumin and its derivatives, is also unrelated to the well researched and described
antioxidant properties of curcumin(oids) and tetrahydrocurcumin(oids). It is a novel
way of accomplishing a broad regulatory effect on biological processes. A
comparable regulatory effect on cross-linking of proteins in the organism was not
described previously with any single compound or with compounded products. In
essence the invention differs from other compounds affecting the cross-linking
process in that the previously described compounds would stimulate or inhibit
intracellular cross-linking, while the invention would act as a cross-linking modifier
(CLM) or "crossregulin". These terms are used here for the first time to describe the
mechanism of the invention. CLM or crossregulin indicates that a composition of the
present invention exerts a positive influence on the post-translational protein
modification and cross-linking process in both physiological and pathological
conditions alike. The CLM effect of the invention is accomplished by using a specific
composition of the product as described in the section "Process of Obtaining THC".
The mechanism also occurs with modifications of the THC molecule, for example
ether THC, trimethyl ether THC, dimethyl ether THC, benzyl ether THC and
potassium, magnesium and copper salts of THC. The mechanism of the invention
has been demonstrated in in vivo experimental conditions such as preventing UV sunburn and cross-linking (percent of thymine dimer cells) in the skin cells exposed
to UV radiation. The biological properties of the invention were compared with those
of curcuminoids, and despite the fact that the present composition is devoid of
yellow color it still produces comparable protective effects against UV radiation as
that of yellow curcuminoids. This activity appears to be different from the antioxidant
effects characteristic of THC and curcuminoids.
The following describes the mechanism of the invention in detail:
1. It is proposed that in physiologic states characterized by optimal post-
translational protein modification, protein cross-linking, as well as the optimal
cell electric potential, THC would not interfere with the above exemplified
states of cellular well-being.
2. However, in the states, characterized by poor morphological and functional
differentiation of cells, e.g. pre-malignant states or malignancy, THC would
contribute to protein post-translational modification and cross-linking and
optimization of cell electric potential. This would be accomplished by
interaction between phenolic rings and olefinic bonds of THC and NH2 as well
as COO groups of the protein amino acids. The resulting bonds
would act as an "anchor" that would initiate cross-linking with other proteins.
3. In the states of cell aging and in the states where random cross-linking is precipitated by a thermal injury due to the action of UV rays, for example, the
undesired process would be prevented by the invention. THC would prevent
undesired cross-linking by attaching to amino and carboxy groups of the
protein amino acids. This would result in interference with the random cross-
linking, a mechanism comparable to a "buffering" action. This "buffering"
action would effectively stop the chain reaction which otherwise would lead to
deterioration of cell homeostasis, premature aging and premature cell death.
4. The invention also has a supportive mechanism in preventing random cross-
linking of intracellular proteins which is based on the antioxidant properties of
THC, specifically on the invention's ability to scavenge free-radicals. It is
proposed that due to this antioxidant action it would facilitate disruption of the
electric charges of "supercharged" protein molecules, which is one of the
reasons why random cross-linking is accelerated. This combined mechanism
of crossregulin and antioxidant action is particularly applicable in preventing
"age spots" due to lipofuscin formation, and inflammatory conditions such as
psoriasis.
5. The source of oxygen moieties in THC for binding to NH2 or COO groups
would be from the para hydroxy group no longer in conjugation with the olefinic
bond, and from the hydroxy groups of phenolic rings. 6. Unlike the curcumin(oid) compound(s), THC molecule(s) can rotate more
freely, thus providing better access to the reactive groups for "anchor" and/or
"buffering" reactions.
7. Whereas, as previously mentioned, the invention is relatively inactive in
states of cell physiology, its crossregulin mechanism would be gradually
expressed proportionate to the degree of stress acting upon the cell. Thus the
crossregulin mechanism of THC in the biological system would depend on the
kind of injurious action experienced by the biological system.
Process of obtaining THC:
Tetrahydro curcuminoids of the invention are products of the chemical
reduction of curcuminoids, obtained from the rhizomes of Curcuma longa, commonly
known as turmeric, or any other suitable plant from the botanical family
Zingiberaceae.
The prior art teaches that when curcuminoids obtained from Curcuma longa
are catalytically reduced and then isolated in the process of crystallization,
tetrahydrocurcumin (THC) is recovered without a loss, but with substantial losses of
tetrahydrobisdemethoxy curcumin (THBDC) and tetrahydrodemethoxy curcumin
(THDC). As a result, the ratio among tetrahydrocurcuminoids is significantly
changed as compared to tile original ratio of parent compounds, curcuminoids. The
ratio of curcuminoids is preferred, because it correlates with the optimal biological activity of curcuminoids. U.S. Patent No. 5,861 ,415 describes curcuminoids in a
specific and optimal ratio as Bioprotectants, providing maximal protective effect of
the living cell.
The present invention provides a new, more efficient method of recovering
THCs. This process prevents the loss of TBDMC and TDMC, and provides a
Bioprotectant composition of THCs.
THCurcuminoids % content prior art % content invention
Tetrahydro curcuminoids 90 75 - 85
Tetrahydro demethoxy curcumin 9 10 - 20
Tetrahydro bisdemethoxy curcumin 1 2 - 4.5
Brief outline of the prior art process for producing THCs
Natural curcuminoids (95 %) - 100 gm
Acetone - 1 liter
Pd-C (catalyst) 5% - 2.5 gm
The solution of natural curcuminoids was charged into a hydrogenator. The
catalyst was added carefully and the pressure of hydrogen was maintained at about
2 Kg/cm2. This reaction is completed in 3 to 4 hrs. The catalyst is then filtered out and removed. The acetone solution is concentrated under vacuum. The residue is
crystallized from toluenes. The tetrahydro compounds are crystallized out as a
creamy white powder having a melting point of 92-94°C.
Process of the Present invention
This process utilizes a novel method of saturation of two olefinic bonds in
curcuminoids in the catalytic hydrogen transfer reaction known as a hydride transfer
reaction. The following is an example of a hydride transfer method.
1. A 3 liter round bottom flask with a stirrer and a reflux condenser is charged
with 1 liter ethyl acetate;
2. The solution is charged with 250 gm of natural curcuminoids in a previously
described Bioprotectant composition, i.e. curcumin (C) 70-80%, Demethoxy
curcumin (DMC) 15-20% and Bisdemethoxy curcumin (BDMC 2.5-6.5% and
stirred to uniformity at room temperature;
3. This mixture is charged with 1 liter of triethylamine followed by 12 gm of
Palladium carbon and 80 ml of formic acid; 4. The mixture was stirred and subsequently refluxed at 80 deg. Celsius for
12 hours.
5. Periodically formic acid in 5 ml aliquots was charged at 2 hours intervals
while a reflux was carried on;
6. TLC was checked to monitor the completion of the reaction, (system: Silica
Precoated 0.25 mm thick plates/chloroform : methanol = 9: 1 );
7. After completion of the reaction, the reaction mixture was cooled to room
temperature. It was filtered to remove palladium catalyst and washed with 50
ml of ethyl acetate;
8. Ethyl acetate and triethylamine were distilled off ( 2 liters of distillate was
collected);
9. The crude mass was then extracted with toluene (3 x 500 ml);
10. The toluene extract was washed with 200 ml of 5% HCI and 3 x 500 ml of
water and dried over sodium sulphate; 11. The solvent was removed under vacuum to get a pale yellow paste
approximately 200 gm.
12. The paste was slurred with 200 ml of ether, filtered and dried under
vacuum to obtain a pale yellow powder of tetrahydrocurcuminoids, Yield 175
gm (70%).
Assay:
THC 75 to 85%
THDHC 10 to 20%
THBDMC 2 to 4.5%
In another variation of hydride transfer reaction, curcuminoids were treated
with ammonium formate or sodium dihydrogen phosphite (as a catalytic hydrogen
transfer reagent) in the presence of Pd/C and acetic acid. The reduction is complete
in 20 to 30 minutes at 100 to 110 Celsius degrees. This method yields 50% of
tetrahydrocurcuminoids.
THC can be administered topically in a suitable vehicle or with an application
device, e.g. a transdermal patch, in a concentration ranging from 0.025% to 5%; in
the form of subcutaneous injections or a transdermal patch in a concentration ranging from 0.025% to 5%; or in orally administered dosage form in a concentration
ranging from 5 mg to 500 mg per dose or 0.083 - 8.3 mg/kg of body weight. THC is
suitable for human and animal use.
Other routes of administration include but are not limited to: aerosol or other
device for delivery to the lungs, nasal spray, intravenous, intramuscular,
intraperitoneal, intrathecal, vaginal, and rectal. Excipients which can be used as a
vehicle for the delivery of the phage will be apparent to those skilled in the art.
Compositions of tetrahydrocurcuminoids for topical administration should be
essentially colorless. The term "essentially colorless" is intended to mean that the
composition contains no color or such a small amount of color that the composition
does not stain the skin upon topical administration.
The term "color removed curcumin" refers to the tetrahydrocurcuminoid
mixture according to the present invention.
THC molecule acting as an "anchor" for amino acids of proteins, to initiate protein crosslinks. This mechanism would contribute to the physiological cross-links.
Figure imgf000017_0001
Legend amino acid/protein
THC molecule acting as a buffer in preventing random cross-links in UN precipitated cross-links. This mechanism would contribute to die prevention of pathological crosslinks.
Figure imgf000017_0002
THC
Legend
-amino acid protein INTRODUCTION
The study was designed to determine the acute oral toxiάty of Colour Removed Curcumin to Sprague Dawiey rats. The guideϋnes followed were as prescribed by die Gaitonde Committee Guidefines, CH, New Delhi
PROJECT N0.4952
MATERIALS AND METHODS
TEST SUBSTANCE
Sponsor Sa i Chemicals and Extracts LtcL, Bangalore.
Label on Sample : Colour Removed Cαrcnmin Batch No.- CRC/65 ARNo. - BL8C 0445
Description Creamy white crystalline powder
Identification IR Spectrum : Complies
Solubility Soluble in methanol
Purity by HPLC 99.0%
PROJECT N0.4 52
SUMMARY
The study now reported was designed to determine die acute oral toxiάty of Colour Removed Curcumin to Sprague Dawiey rats. The test substance suspended in co oil was administered by oral route to rats. The rats were observed for 14 days after treatment for the product related symptoms. The test snbstance caused salivation all animals and polyurea in two animals with onset at 2 hours after the treatment. AH *t n_. survived through the study period of 14 days and were free of intoxicating signs 24 hours after the treatment.
The LD50 value of Colour Removed Curcumin in rats by oral route was found to be greater than 5000 mg/kg. TEST SYSTEM
Species Rat
Strain Sprague Dawiey
Source LIT. Animal house
Sex Male and Female
Age 6 to 8 weeks
Weight range 120.5 to 141.9 gms
No. of animals per dose 5 per sex
Randomization Randomly selected in groups of five of like sex at the time of initiating the study.
Accfimatiσn Not done as the animals were bred and reared in die same Institute.
Identification of antmqls By cage number and individual marking on for. Diet Pelleted feed supplied by Nav Maharashtra Chakan Oil Mills Ltd., Pune.
Water Aquaguard pure water in glass bottles ad libitum.
Management The rats were housed S each, of the same sex in polypropylene cages provided with bedding of husk. The temperature was maintained between 20 &-24 °C and relative humidity between 50 and 60%; 12 hours each of dark and light cycle was maintained.
Route Oral gavage
Vehicle used Com oil
Dose volume 10 ml/kg The test substance, suspended in coin oil was administered once each by oral gxvage in rats of both saxes. The rats were starved for 16 hours before and 2 hours after the administration of the test substance. The animals woe observed for 14 days after the dosing for the product related toxic symptoms and mortality. Necropsy was carried out animals died during the observation period. At the end of the observation period the surviving test animals were sacrificed, dissected and examined for gross pathological abnormality, if any.
Figure imgf000020_0001
Ten rats (5 male and 5 female) was allocated to treatment as follows.
Group No._ No.of rats per sex Dosc ft/ty
1 5000
PROJECT NO.4 52
RESULTS
LDso in rats
Group No. Dose (mg kg) No. of aiiml. _ Mortality
No. of animals treated
5000 0/10
Symptoms :
Group 1 Salivation in all animals and polyurea in two animals were observed with onset at 2 hours after the treatment. All animals survived through the study period of 14 days and were free of intoxicating signs 24 hours after the treatment.
Necropsy findings : No significant changes could be seen with naked eye.
Figure imgf000020_0002
SUMMARY
The study was deagned to determine the primary skm irritation potential of Colour Removed Cuxαmin supplied by Sami Chemicals and Extracts Ltd* Bangalore. The test substance in die amount of 500 mg was applied to shorn back dm both intact and abraded site of three rabbits per sex. Each site of application was carefhHy observed and the reaction evaluated according to Drake's method at 24 and 72 hours. Colour Removed Curcumin did not cause irritation to skin in rabbits. Primary skin irritation score - 0.00.
MATERIALS AND METHODS
TEST SUBSTANCE
Sponsor Sami Chemicals and Extracts Ltd*, Bangalore.
Label on Sample : Colour Removed Curcumin Batch No.- CRC/65 ARNo. - BL8C 0445
Description Creamy white crystalline powder
Identification IR Spectrum : Complies
Solubility : Soluble in methanol
Purity by HPLC : 99.0% TEST SYSTEM
Species Rabbit
Strain New Zealand White
Source Serum Institute of India Ltd., Pane.
Sex Male and Female
Age Healthy adult
Weight range 2.00 to 2.40 kg
No. of animals per dose 3 per sex
Randomization Randomly selected in groups of three of like sex at the time of initiating the study.
Acclimation Seven days prior to treatment.
Identification of animals By cage number and individual marking on ear.
Diet Pelleted feed supplied by Nav Maharashtra Chakan Oil Mills Ltd., Pone.
Water Aquaguard pure water in glass bottles ad libitum.
Management The rabbits were housed individually each, in stainless steel cages provided with stainless steel mesh bottom. The temperature was maintained between 20 & 23 "fiend relative humidity between 50 and 60 ; 12 hours each of dark and light cycle was maintained.
Route Dermal
Dose volume 500 tog/she. EXPERIMENTAL PROCEDURE
Three rabbits per sex were used for this study. The hair on both die sides were removed by electric clippers one day before the treatment The test snfwtmrr in die amount of 500 mg was applied to the intact and abraded slrin sites of each test animaL Care was taken to note that the abrasions penetrate die Stratum coπieum but not he dezmis. A gauze patch was secured over each treated area by means of adhesive tape. The animals were housed singly with plastic collar around their necks for 24 hours α order to avoid die mgestion of test substance. The patch and unabsorbed test substance was removed after 24 hours. Each site of application was carefully observed and reaction evaluated according to Draize's method at 24 and 72 hours.
Braize Evaluation of Skin Reactions -
Grading of skin reaction
1. Erythema and Eschar Formation
No erythema 0
Very slight erythema (barely perceptible) I
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4
Total n -r miiTT- possible erythema scor : 4
2. Edema Formation
No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well defined by definite raising) 2
Moderate edema (raised approximately 1 mm 3
Severe edema (raised more than 1 mm and extending beyond area of exposure) 4
Total maximum possible edema score : 4 RESULTS
Summary of mdividual dermal irritation score following appScatiøn of Colour Removed Curcumin on rabbits.
Figure imgf000024_0002
Primary skin irritation score 0.00 / 4 = 0.00.
CONCLUSION
It is concluded based on the presem study that of the test substance Colour Removed Curcumin supplied by Sami Chemicals and Extracts Ltd., Bangalore did not cause irritation to skin in rabbits. Primary skin irritation score : 0.00.
Figure imgf000024_0001
Figure imgf000025_0001
*NTC: natural STC: synthetic.
O 00/61162
Effect of topical application of curcumin and tetrahydrocurcumin on ultraviolet B light-induced formation of thymine dlmers in mouse epidermis
Treatment Number of Percent of Percent of per group thymine dimer cells protection
1. No UVB 3 0
2. Acetone +UVB 7 8.9 ± 0.02 0
3. THC (10 μ ol) + UVB β 7.3 * 0.03 24.7
4. THC (20 μmol) + UVB 6 1.6 ± 1.30 82.0 δ. Curcumin (10 μmol) + UVB 5 1.6 ± 0.01 82.0 6. Curcumin (20 μmol) + UVB 5 1.2 * 1.69 86.5
Female SKH-1 mice (8-9 weeks old) were treated topically with 100 μl acetone, or test compound in 100 μl acetone. Five minutes later, the mice were irradiated with a βlnglβ dose of UVB (30 mJ/cm2). The mice were killed one hour later, and the skin samples were stored in a 10% formalin-phosphate buffer for thymine dimβrs assay.
Effect of UVB-lnducβd formation of skin sunburn cells in mouse epidermis
Treetmeπt Number of Percent percent per group of sunburn cells of inhibition
No UVB 0.1
Aoetone + UVB (60 mJ/cm2)
DB + (10 μmol) 5 1 .2 60 DBM + (20 μmol) 5 0.9 70
Tetrahydrocurcumin (10 μmol) UVB 5 1 .6 47 Tetrahydrocurcumin (20 μmol) UVB 5 1 .4 53
Female Sencar mice were irradiated with UVB (80 mJ/cn.2), The mice were treated topically with 100 μl acetone or inhibitor in 100 μl aoetone at 5 min before the UVB irradiation. Eight hours later, the mice were killed and skin samples were removed and kept in 10 % formalin phosphate buffer for sunburn cells assay.

Claims

1. A method for regulating random intracellular protein cross-linking and
optimizing cell electric potential in a patient, comprising administering an effective
amount of a mixture of tetrahydrocurcuminoids to a patient in need of such
regulation.
2. The method according to claim 1 , wherein said random intracellular protein
cross-linking is due to exposure to UV rays.
3. The method according to claim 1 , wherein said random intracellular protein
cross-linking is due to the presence of a malignancy.
4. The method according to claim 1 , wherein said intracellular protein
crosslinking is regulated by bonding said tetrahydrocurcuminoids to amino and
carboxy groups of amino acids.
5. A method for treating a premalignant state or a malignancy in a patient
comprising administering an effective amount of a mixture of tetrahydrocurcuminoids
to a patient suffering from a premalignant state or a malignancy.
6. The method according to claim 5, wherein said malignancy is melanoma.
7. A method for increasing intracellular transglutaminase activity in a cell
comprising administering an effective amount of a mixture of
tetrahydrocurcuminoids to a cell with decreased intracellular transglutaminase
activity.
8. A method for producing tetrahydrocurcuminoids by saturating two olefinic
bonds in a mixture of curcuminoids, comprising the steps of
mixing ethyl acetate and curcuminoids at room temperature;
adding a catalytic hydrogen transfer reagent followed by palladium carbon
and an acid;
stirring and subsequently refluxing the mixture while periodically adding
aliquots of said acid;
cooling the resulting mixture to room temperature;
filtering the mixture to remove any palladium carbon;
washing the mixture in ethyl acetate;
distilling off any ethyl acetate;
extracting the resulting crude mass with toluene to produce a toluene extract;
washing the toluene extract with HCI and water;
drying said extract;
removing any remaining toluene under vacuum to produce a paste;
mixing said paste with ether to produce a slurry, and filtering and drying said slurry under vacuum to obtain tetrahydrocurcuminoids.
9. The method according to claim 8, wherein said curcuminoids comprise
70-80% curcumin, 15-20% demethoxy curcumin, and 2.5-6.5% bisdemethoxy
curcumin.
10. The method according to claim 8, wherein said catalytic hydrogen transfer
reagent is selected from the group consisting of triethylamine, ammonium formate
and sodium dihydrogen phosphite.
11. The method according to claim 8, wherein said acid is selected from the
group consisting of formic acid and acetic acid.
12. A method for producing tetrahydrocurcuminoids by saturating two olefinic
bonds in a mixture of curcuminoids, comprising the steps of
mixing ethyl acetate and curcuminoids at room temperature;
adding a catalytic hydrogen transfer reagent followed by palladium carbon and
an acid;
heating the resulting mixture;
cooling the heated mixture to room temperature;
filtering the mixture to remove any palladium carbon; and recovering any tetrahydrocurcuminoids.
13. A mixture comprising 75-85% tertahydrocurcumin, 10-20%
tetrahydrodemethoxy curcumin, and 2.0-4.5% tetrahydrobisdemethoxy curcumin,
wherein said mixture is substantially colorless.
14. The mixture according to claim 13, wherein said mixture is in a form suitable
for topical administration.
15. A method for reducing "age spots" due to lipofuscin formation in a patient in
need of such treatment, comprising administering an effective amount of a mixture
comprising 75-85% tertrahydrocurcumin, 10-20% tetrahydrodemethoxy curcumin,
and 2.0-4.5% tetrahydrobisdemethoxy curcumin to said patient.
16. The method according to claim 13, wherein said composition is administered
topically.
17. A colorless mixture comprising tertrahydrocurcumin, tetrahydrodemethoxy
curcumin, and tetrahydrobisdemethoxy curcumin.
PCT/US2000/008711 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa WO2000061162A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US12854099P true 1999-04-09 1999-04-09
US60/128,540 1999-04-09

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
CA002369381A CA2369381A1 (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
EP00921582A EP1171144B1 (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
AU41879/00A AU4187900A (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
NZ514884A NZ514884A (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
DK00921582T DK1171144T3 (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids for the regulation of physiological and pathological conditions in the skin and mucous membranes
JP2000610495A JP2002541205A (en) 1999-04-09 2000-04-07 Methods of using tetrahydrocurcuminoids to control physiological and pathological events involving skin and mucous membranes and methods of making the same
DE60033383T DE60033383T2 (en) 1999-04-09 2000-04-07 USE OF TETRAHYDROCURCUMINOIDS FOR THE REGULATION OF PHYSIOLOGICAL AND PATHOLOGICAL CASES IN THE SKIN AND MUCOSA
US09/972,150 US6653327B2 (en) 1999-04-09 2001-10-09 Cross-regulin composition of tumeric-derived tetrahydrocurcuminoids for skin lightening and protection against UVB rays

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US09/972,150 Continuation-In-Part US6653327B2 (en) 1999-04-09 2001-10-09 Cross-regulin composition of tumeric-derived tetrahydrocurcuminoids for skin lightening and protection against UVB rays

Publications (2)

Publication Number Publication Date
WO2000061162A1 true WO2000061162A1 (en) 2000-10-19
WO2000061162A9 WO2000061162A9 (en) 2001-11-29

Family

ID=22435823

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/008711 WO2000061162A1 (en) 1999-04-09 2000-04-07 Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa

Country Status (11)

Country Link
EP (1) EP1171144B1 (en)
JP (1) JP2002541205A (en)
AT (1) AT353662T (en)
AU (1) AU4187900A (en)
CA (1) CA2369381A1 (en)
DE (1) DE60033383T2 (en)
DK (1) DK1171144T3 (en)
ES (1) ES2282101T3 (en)
NZ (1) NZ514884A (en)
PT (1) PT1171144E (en)
WO (1) WO2000061162A1 (en)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1108419A1 (en) * 1999-12-14 2001-06-20 Avon Products, Inc. Cosmetic composition and methods of use
WO2002032415A2 (en) * 2000-10-19 2002-04-25 Sabinsa Corporation Process of making and method of use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa cells
DE10121069A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Use of tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin to prepare a cosmetic or dermatological composition for treating or preventing skin aging
DE10121093A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin is/are used in production of cosmetic or dermatological skin barrier preparations
DE10121067A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Cosmetic or dermatological preparations acting against skin pigmentation or tanning contain synergistic combinations of L-tyrosine sulfate or tyrosine O-sulfate ester with curcuminoids
DE10121090A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag New cosmetic or dermatological compositions comprise a synergistic combination of sericoside and tetrahydrocurcumin or its derivatives, useful for treating e.g. skin inflammation and aging symptoms
DE10121089A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Cosmetic or dermatological compositions, useful against, e.g. skin inflammation, containing synergistic combination of nitrogen monoxide synthase inhibitor and tetrahydrocurcumin or derivative
DE10121070A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Use of tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin to prepare a cosmetic or dermatological composition for stimulating skin ceramide biosynthesis
US20110077275A1 (en) * 2008-11-26 2011-03-31 Jan Zielinski Antioxidant compositions for soft oral tissue and methods of formulation and use thereof
WO2013049507A1 (en) 2011-09-30 2013-04-04 Perio Sciences, Llc Antioxidant compositions for treatment of inflammation or oxidative damage
FR2983402A1 (en) * 2011-12-05 2013-06-07 Oreal CAPILLARY COLORING COMPOSITION COMPRISING A TETRAHYDROCURCUMINOID, A CAPILLARY COLOR, AND A LIQUID MONOALCOHOL

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106266988A (en) * 2016-09-12 2017-01-04 四川聚豪生物科技有限公司 A kind of for Chinese medicine composition treating goiter due to disorder of QI and preparation method thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5266344A (en) * 1988-08-12 1993-11-30 Kabushiki Kaisha Kobe Seiko Sho Method for making tetrahydrocurcumin and a substance containing the antioxidative substance tetrahydrocurcumin
JPH06128133A (en) * 1992-10-20 1994-05-10 Kobe Steel Ltd External agent for preventing ultraviolet hazard
WO1995018606A1 (en) * 1994-01-06 1995-07-13 Research Development Foundation Curcumin, analogues of curcumin and novel uses thereof
WO1997003674A1 (en) * 1995-07-14 1997-02-06 Sabinsa Corporation Bioprotectant composition, method of use and extraction process of curcuminoids

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5266344A (en) * 1988-08-12 1993-11-30 Kabushiki Kaisha Kobe Seiko Sho Method for making tetrahydrocurcumin and a substance containing the antioxidative substance tetrahydrocurcumin
JPH06128133A (en) * 1992-10-20 1994-05-10 Kobe Steel Ltd External agent for preventing ultraviolet hazard
WO1995018606A1 (en) * 1994-01-06 1995-07-13 Research Development Foundation Curcumin, analogues of curcumin and novel uses thereof
WO1997003674A1 (en) * 1995-07-14 1997-02-06 Sabinsa Corporation Bioprotectant composition, method of use and extraction process of curcuminoids

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
MENON L G ET AL: "Anti-metastatic activity of curcumin and catechin.", CANCER LETTERS, (1999 JUL 1) 141 (1-2) 159-65., XP000933941 *
MENON L G ET AL: "Inhibition of lung metastasis in mice induced by B16F10 melanoma cells by polyphenolic compounds.", CANCER LETTERS, (1995 AUG 16) 95 (1-2) 221-5., XP000934170 *
MULKY, N. ET AL: "Antimutagenicity of curcumins and related compounds: the structural requirement for the antimutagenicity of curcumins", INDIAN DRUGS (1987), 25(3), 91-5, XP000934197 *
NAGABHUSHAN, M. ET AL: "Curcumins as inhibitors of nitrosation in vitro", MUTAT. RES. (1988), 202(1), 163-9, XP000933930 *
OSAWA, TOSHIHIKO ET AL: "Antioxidative activity of tetrahydrocurcumin", INT. CONGR. SER. - EXCERPTA MED. (1992), 998(OXYGEN RADICALS), 801-4, XP000933940 *
RUBY, A. J. ET AL: "Antitumor and antioxidant activity of natural curcuminoids", CANCER LETT. (SHANNON, IREL.) (1995), 94(1), 79-83, XP000933937 *
SHARMA, O. P.: "Antioxidant activity of curcumin and related compounds", BIOCHEM. PHARMACOL. (1976), 25(15), 1811-12, XP000933938 *
WALKER, M. J. ET AL: "Curcumin in a chemoprevention model of melanoma.", PROCEEDINGS OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH ANNUAL MEETING, (MARCH, 1998) VOL. 39, PP. 19. MEETING INFO.: 89TH ANNUAL MEETING OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH NEW ORLEANS, LOUISIANA, USA MARCH 28-APRIL 1, 1998 AMERICAN, XP000929727 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6521668B2 (en) 1999-12-14 2003-02-18 Avon Products, Inc. Cosmetic composition and methods of use
EP1108419A1 (en) * 1999-12-14 2001-06-20 Avon Products, Inc. Cosmetic composition and methods of use
WO2002032415A3 (en) * 2000-10-19 2003-04-17 Sabinsa Corp Process of making and method of use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa cells
WO2002032415A2 (en) * 2000-10-19 2002-04-25 Sabinsa Corporation Process of making and method of use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa cells
DE10121093A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin is/are used in production of cosmetic or dermatological skin barrier preparations
DE10121067A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Cosmetic or dermatological preparations acting against skin pigmentation or tanning contain synergistic combinations of L-tyrosine sulfate or tyrosine O-sulfate ester with curcuminoids
DE10121090A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag New cosmetic or dermatological compositions comprise a synergistic combination of sericoside and tetrahydrocurcumin or its derivatives, useful for treating e.g. skin inflammation and aging symptoms
DE10121089A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Cosmetic or dermatological compositions, useful against, e.g. skin inflammation, containing synergistic combination of nitrogen monoxide synthase inhibitor and tetrahydrocurcumin or derivative
DE10121070A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Use of tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin to prepare a cosmetic or dermatological composition for stimulating skin ceramide biosynthesis
DE10121069A1 (en) * 2001-04-26 2002-10-31 Beiersdorf Ag Use of tetrahydrocurcumin, tetrahydrodemethoxycurcumin and/or tetrahydrobisdemethoxycurcumin to prepare a cosmetic or dermatological composition for treating or preventing skin aging
US20110077275A1 (en) * 2008-11-26 2011-03-31 Jan Zielinski Antioxidant compositions for soft oral tissue and methods of formulation and use thereof
WO2013049507A1 (en) 2011-09-30 2013-04-04 Perio Sciences, Llc Antioxidant compositions for treatment of inflammation or oxidative damage
US9421180B2 (en) 2011-09-30 2016-08-23 Perio Sciences, Llc Antioxidant compositions for treatment of inflammation or oxidative damage
US10918613B2 (en) 2011-09-30 2021-02-16 Perio Sciences, Llc Antioxidant compositions for treatment of inflammation or oxidative damage
FR2983402A1 (en) * 2011-12-05 2013-06-07 Oreal CAPILLARY COLORING COMPOSITION COMPRISING A TETRAHYDROCURCUMINOID, A CAPILLARY COLOR, AND A LIQUID MONOALCOHOL
WO2013083388A1 (en) 2011-12-05 2013-06-13 L'oreal Hair dyeing composition comprising a tetrahydrocurcuminoid, a hair dye and a liquid monoalcohol

Also Published As

Publication number Publication date
DE60033383T2 (en) 2007-05-31
JP2002541205A (en) 2002-12-03
WO2000061162A9 (en) 2001-11-29
NZ514884A (en) 2008-05-30
AT353662T (en) 2007-03-15
AU4187900A (en) 2000-11-14
DK1171144T3 (en) 2007-06-11
EP1171144B1 (en) 2007-02-14
PT1171144E (en) 2007-04-30
DE60033383D1 (en) 2007-03-29
ES2282101T3 (en) 2007-10-16
CA2369381A1 (en) 2000-10-19
EP1171144A1 (en) 2002-01-16

Similar Documents

Publication Publication Date Title
JP2019112422A (en) Melanin modification compositions and methods of use
EP1328263B1 (en) Process of making and method of use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa cells
EP1171144B1 (en) Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
US8492578B2 (en) Compounds useful in therapeutic and cosmetic methods
NZ207686A (en) Phosphorylated derivatives of 3-(3,4-dihydroxy) -phenyl-alanine and compositions for increasing skin melanisation
DE2817133C2 (en)
Hussain et al. Fabrication of anti-vitiligo ointment containing Psoralea corylifolia: in vitro and in vivo characterization
JPH11503111A (en) Treatment of epithelial hyperplasia by topical application of hydroxylated aromatic protein-crosslinking compounds
KR102156731B1 (en) Bakuchiol compositions for treatment of post inflammatory hyperpigmentation
EP2762131B1 (en) Use of Salvia Haenkei extracts in compositions for antisenescence
VON et al. PATHOLOGICAL EVENTS IN THE SKIN AND MUCOSA
EP0526502B1 (en) Phenolic amine depigmenting and antimelanoma agents
AU2006235807B2 (en) Use of tetrahydrocurcuminoids to regulate physiological and pathological events in the skin and mucosa
KR20010079933A (en) Use of a boldo extract in a cosmetic or dermatological product
DE2616478A1 (en) MEDICINAL PRODUCTS AND COSMETICS CONTAINING FLUORACYL RESORCINES
ZUR et al. REGULATE PHYSIOLOGICAL AND PATHOLOGICAL EVENTS IN THE SKIN AND MUCOSA CELLS
KR100975078B1 (en) A Cosmetic composition containing Azolla imbricata extract
US20160106661A1 (en) Screening method and substances for contrasting aging
US20210113523A1 (en) Compositions For Treating And/Or Preventing Photodynamic Therapy Side Effects
KR20030011980A (en) Hyperpigmentation treatment composition containing stilbene derivatives as an effective component
Romiszewska et al. The use of 5-aminolevulinic acid and its derivatives in photodynamic therapy and diagnosis
KR100278747B1 (en) Whitening cosmetic compositions containing ascorbic acid-aminopropanol phosphoric acid diester
CN110615826A (en) Novel compound, cosmetic composition and pharmaceutical composition containing same
WO2021133307A1 (en) Cosmetic formulation containing the smoke tree extract
JPH08231368A (en) Light-aging preventing agent and skin cosmetic containing the agent

Legal Events

Date Code Title Description
AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase in:

Ref document number: 2369381

Country of ref document: CA

Ref country code: JP

Ref document number: 2000 610495

Kind code of ref document: A

Format of ref document f/p: F

Ref country code: CA

Ref document number: 2369381

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 514884

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 2000921582

Country of ref document: EP

AK Designated states

Kind code of ref document: C2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

COP Corrected version of pamphlet

Free format text: PAGES 1-24, DESCRIPTION, REPLACED BY NEW PAGES 1-24; DUE TO LATE TRANSMITTAL BY THE RECEIVING OFFICE

AL Designated countries for regional patents

Kind code of ref document: C2

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

WWP Wipo information: published in national office

Ref document number: 2000921582

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWG Wipo information: grant in national office

Ref document number: 2000921582

Country of ref document: EP

DPE2 Request for preliminary examination filed before expiration of 19th month from priority date (pct application filed from 20040101)