UTILISATION D'UN EXTRAIT PARTIEL OU COMPLET DE CAMELLIA SINENSIS L. NON FERMENTE POUR LA PREPARATION D'UN MEDICAMENT, D'UN PRODUIT DE SOINS MEDICAUX, D'UNE PREPARATION COSMETIQUE OU D'UN PRODUIT ALIMENTAIRE COMPLEMENTAIRE

USE OF A PARTIAL OR COMPLETE EXTRACT OF NOT FERMENTED CAMELLIA SINENSIS L. FOR THE PREPARATION OF A MEDICAMENT, A MEDICAL CARE PRODUCT, A COSMETIC PREPARATION OR A FOOD COMPLEMENTARY PRODUCT

Abrégé français

La présente invention concerne l'utilisation d'un extrait partiel ou complet de Camellia sinensis L. non fermenté pour la préparation d'un médicament, d'un produit de soins médicaux, d'une préparation cosmétique ou d'un produit alimentaire complémentaire; ces préparations préviennent, ou du moins réduisent considérablement, la formation de nécroses et/ou d'atrophies dans les tissus humains ou animaux et/ou la mort prématurée des cellules et tissus/colonies cellulaires vascularisés et non vascularisés dans l'organisme humain ou animal, et non seulement elles favorisent l'adhésion entre des cellules uniques ou groupes de cellules appartenant à des types de tissus identiques, mais elles empêchent également, ou du moins réduisent considérablement, l'adhésion entre des cellules uniques ou des groupes de cellules appartenant à des types de tissus différents non histocompatibles.

Abrégé anglais

The present invention is directed to the use
of a partial or complete extract of not fermented
Camellia sinensis L. for the preparation of a medicament, a
medical care product, a cosmetic preparation or a food
complementary product, whereby these preparations
- prevent or at least reduce considerably the
formation of necrosis and/or atrophies in human or
animal tissues and/or the premature mortification of
vascularized and non-vascularized cells and cellular
tissues/colonies in the human or animal body, and
- not only promote the adhesion between
single, to the same tissue type belonging cells or cell
unions,
- but also prevent or at least reduce
considerably the adhesion between single, to different not
histo-compatible tissue types belonging cells or cell
unions.

Revendications

13
Claims
1. Use of a partial or complete extract of un-
fermented Camellia sinensis L. for the preparation of a
medicament, a medical care product, a cosmetic prepara-
tion or a food complementary product, whereby these pre-
parations
- prevent or at least reduce considerably the
formation of necrosis and/or atrophies in human or ani-
mal tissues and/or the premature mortification of vascu-
larized and non-vascularized cells and cellular tissu-
es/colonies in the human or animal body, and
- not only promote the adhesion between sing-
le, to the same tissue type belonging cells or cell uni-
ons,
- but also prevent or at least reduce consi-
derably the adhesion between single, to different not
histo-compatible tissue types belonging cells or cell
unions.
2. Use according to claim 1, characterized in
that the medicament or the medical care product is in a
pharmaceutically acceptable administrative form.
3. Use according to claim 2, characterized in
that the pharmaceutical acceptable administrative form
is a
- solid administrative form for oral applica-
tion,

14
- liquid administrative form for oral, paren-
teral, rectal, vaginal and tropic application,
- semisolid administrative form for topic,
oral, rectal and vaginal application.
4. Use according to claim 3, characterized in
that the solid administrative form is a tablet, a film-
tablet, a dragee, a pellet, a hard gelatin-capsule or a
soft gelatin-capsule.
5. Use according to claim 3, characterized in
that the liquid administrative form is a dropping solu-
tion, a spray, an injection solution or a syrup.
6. Use according to claim 3, characterized in
that the semisolid administrative form is a cream, a
gel, an ointment, a paste or a suppository.
7. Use according to claim 1, characterized in
that the cosmetic preparation or the food complementary
product is in the form of a solution, a spray, a cream,
a gel, an ointment, a paste, a dragee, a capsule, an am-
pule or a shampoo.
8. Use according to any one of claims 1 to 7, cha-
racterized in that the partial or complete extract is
incorporated into nano-capsules or into liposomes.
9 . Use according to any one of clam 1 to 8, cha-
racterized in that said preparations contain additional-
ly at least one additive and/or auxiliary agent.
10. Use according to claim 9, characterized in
that, the additive and/or auxiliary agent is selected

15
from the group, consisting of emulsifiers, stabilizers,
antioxidants, dyestuffs, aromas, disintegration agents,
solvents, lubricants and surfactants.
11. Use according to any one of claims 1 to 10,
characterized in that the partial or complete extract is
contained in an amount from 0.1 % by weight to 95 % by
weight, referred to the total weight of the preparation.
12. Use according to any one of claims 1 to 11,
characterized in that the partial or complete extract
contains besides native polyphenols and purine alkaloids
additionally at least one further native compound, se-
lected from the group consisting of glutamic acid ethyl-
amide, glutamic acid, including their physiologically
salts, and glutamine.
13. Use according to any one of claims 1 to 12,
characterized in that said preparations serve for the
prevention, treatment or post-treatment of
- cachectic states, caused by small intestine
villus atrophies,
- malabsorptions, caused by small intestine
villus atrophies,
- polyneuropathias, caused by non-inflammatory
axon damages,
- parodontosis, caused by gingiva cell growth
disorders and cell-cell-adhesion disorders,
- loss of hair, caused by hair root atrophies,

16
- hypertrophic skin changes,
- hyperplasias, caused by pathological cell
propagation,
- leukozytes-maturation disorders,
- exsudative forms of the tuberculosis of the
lungs,
- zirrhosis of the lungs,
- zirrhosis of the liver,
- emphysemas of the lungs, or
- erythematodes visceralis.
14. Use according to claim 13, characterized
in that the hypertrophic skin changes are psoriasis vul-
garis, neurodermitis, caused by cell-cell-adhesion di-
sorders and lacks of differentiation.
15. Use according to claim 13, characterized
in that the hyperplasias are hypersplenism, caused by
lymphogranulomatosis, foveolic hyperplasia of the ga-
stric mucosa.
16. Use according to claim 13, characterized
in that the leukozytes-maturation disorders are leukae-
mias of the subspecies
a.) acute lymphatic leukaemia,
b.) chronic lymphatic leukaemia,

17
c.) acute myeloic leukaemia, or
d.) chronic myeloic leukaemia.
17. Use according to claim 16, characterized
in that the acute myeloic leukaemia is of the peroxida-
se-type.
18. Use according to claim 13, characterized
in that the zirrhosis of the lungs is chronic intersti-
tial fibrosis with parenchyma depletion.
19. Use according to claim 13, characterized
in that the zirrhosis of the liver is a consequence of
chronic hepatitis.
20. Use according to any one of claims 1 to 19,
characterized in that said preparations serve for the
treatment of human or animal cell and/or tissue cultures
as well as organs outside the human or animal body.
21. Use according to claim 20, characterized
in that said preparations serve for the cultivation and
propagation and/or the three-dimensional reconstruction
of cartilage cells, gingiva cells, hair root cells, skin
cells and skin tissues, retina cells and retina tissues,
heart muscle cells and heart muscle tissues, liver cells
and liver tissues.

Description

CA 02217227 1997-10-24
., 1
Use of a partial or complete extract of not fermented
Camellia sinensis L. for the preparation of a medica-
ment, a medical care product, a cosmetic preparation or
a food complementary product
The present invention is directed to the use
of a partial or complete extract of not fermented Camel-
lia sinensis L. for the preparation of a medicament, a
medical care product, a cosmetic preparation or a food
complementary product.
Preparations of Camellia sinensis L. for medi-
cal and cosmetic applications are known; see "Hagers
Handbuch der Pharmazeutischen Praxis", Vol. 4, Drogen A-
D, Springer-Verlag, (1992), pages 628 - 640.
Due to its content compounds Camellia sinensis
L. has a central stimulating, moderate diuretic, in de-
pendence of the extraction time more or less strong con-
stipatory/anti-diarrhoeal, the heart activity promoting
and possibly antiarteriosclerotic effect, see Stagg
G.V., Millin D.J., (1975), J. Sci. Food. Agric. 26, pa-
ges 1439 - 1459.
The further prior art concerning Camellia si-
nensis L. is described in Zeitschrift fur Phytotherapie
17, (1995), pages 231 - 250.
So are especially assigned to the in Camellia
sinensis L. contained polyphenols an antioxidant effi-
cacy, which shall protect the human body from so called
radicals.
The polyphenols are also assigned to have an
anti-inflammatory efficacy.

CA 02217227 1997-10-24
An inhibition of the tumor formation by means
of Camellia sinensis L. extracts has been proved in ani-
mal experiments and is supported by epidemiological stu-
dies.
Also mentioned are virostatic and bacteriosta-
tic effects of Camellia sinensis L. extracts.
In JP 07/101 837 is described an agent having
anti-dandruff activity. As active component this agent
contains an alcoholic extract of not further specified
tea leaves.
In JP 06/056 687 is described an agent with
which is removed dental tartar, and with which the for-
mation of dental tartar is prevented.
As active components this agent contains an
aqueous or a with a hydrophilic organic solvent obtained
extract of tea leaves (Camellia sinensis).
For the claimed activity has been made respon-
sible the strong antimicrobial effect against periodon-
totic pathogenic bacterias.
In JP 08/073 350 is described an agent for the
improvement of cerebral functions.
As active component is mentioned theanine
(glutamic acid ethylamide). This component may be con-
tained in tea extracts.
In JP 06/100 442 is described an anti-stress
agent for the prevention or mitigatlon of mental and
physical diseases due to stress.

CA 02217227 1997-10-24
As active component this agent contains L-
theanine of native origin as such or in the form of a
pharmaceutically acceptable salt, for example the hydro-
chloride.
5In US 5 071 653 are described processes for
the preparation of extracts of Camellia sinensis.
These extracts promote the growth of bifido-
bacterias.
These extracts contain in contrary to tea in-
fusions, partial and complete extracts of not fermented
Camellia sinensis neither flavone glucosides nor poly-
phenols, and also no therpenoides and no lipophilic com-
pounds.
With these in US 5 071 653 described processes
are made highly selective fractionations of Camellia si-
nensis content compounds, which have a completely unu-
sual activity spectra, that is the promotion of the
growth of intestine bacterias.
Quite surprisingly were found new use possibi-
lities of partial or complete extracts of not fermentedCamelia sinensis L.
The present invention is directed to the use
of a partial or complete extract of not fermented Camel-
lia sinensis L. for the preparation of a medicament, a
medical care product, a cosmetic preparation or a food
complementary product, whereby these preparations
- prevent or at least reduce considerably the
formation of necrosis and/or atrophies in human or ani-

CA 02217227 1997-10-24
mal tissues and/or the premature mortification of vascu-
larized and non-vascularized cells and cellular tissu-
es/colonies in the human or animal body, and
- not only promote the adhesion between sing-
le, to the same tissue type belonging cells or cell uni-
ons,
- but also prevent or at least reduce consi-
derably the adhesion between single, to different not
histo-compatible tissue types belonging cells or cell
unions.
Preferred embodiments of this invention are
defined in the dependent claims.
The extract of Camellia sinensis L. used for
the examinations described below was prepared as fol-
lows.
6 kg dried, not fermented Camellia sinensis L.
(folia) were extracted under stirring with 60 kg of a
mixture of 8 parts by weight of ethanol and 2 parts by
weight of water at a temperature from 25~C to 35~C du-
ring 2 hours.
Then was filtered, and the solvent mixture was
evaporated at a pressure from 50 mbar to 150 mbar and a
temperature from 30~C to 40~C up to a complete evapora-
tion of ethanol.
From the so obtained concentrate were removed
according to EP 0 730 830 A1 the undesired lipophilic
contaminations and residues.

CA 02217227 1997-10-24
The so purified extract was then subjected to
a process for a decrease of the bacterial count (30 se-
conds at a temperature of 120~C).
Then this extract was spray dried.
There was obtained 1 kg of native dry extract
having the following analysis datas.
50 % m/m phenylchroman derivatives, calculated
as epicatechin (HPLC),
6 % m/m caffeine (HPLC)
1 % m/m theobromine (HPLC).
In addition were detected qualitatively:
glutaminic acid-ethylamide
flavonoides and
plant acids (HPTLC).
This product is obtainable from the company
Emil Flachsmann AG in CH-8820 Wadenswil / Switzerland
under the denotation "EFLA 85942".
This product was tested on its effects on a
special cell model, that is the multicellular spheroids.
These spheroids are ball-shaped cell aggrega-
tes, which contain in suspension culture at a diameter
of about 1 mm up to 100'000 cells, and as a rule reflect

CA 02217227 1997-10-24
better the biological relations of cell unions in vivo
than conventional monolayer cultures.
Experiment 1
Tested was the influence of the product "EFLA
85942" onto the volume growth of these spheroids as a
function of the cultivation length and at different con-
centrations of "EFLA 85942" in comparision to control
cultures.
The following results were obtained.
As expected "EFLA 85942" effected a systematic
and in large ranges of the growth length a significant
reduction of the volume growth of the spheroids.
But quite surprisingly no concentration depen-
dence could be detected thereby after the beginning of
the effect over a broad concentration range.
This effect points to a high therapeutical
width.
In addition it was noted quite surprisingly
that the with "EFLA 85942" treated spheroides formed no
noteworthy necrosis during the whole growth phase.
In control cultures which were not treated
with "EFLA 85942" already at diameters of about 200
micrometers were detected central necrosis which in-
creased strongly during the growth phase.

CA 02217227 1997-10-24
At the end of the duration of test the untrea-
ted spheroids showed only a very thin vital border lay-
er.
This occurance of necrosis is typical for the
used experimental model.
This behaviour of the with "EFLA 85942" trea-
ted spheroids has not yet been observed.
It is persumed that the known antioxidative
effect of the polyphenols can not be responsible alone
for this behaviour.
On behalf of this may be used also the res~lts
which were obtained from accompanying investigations on
singly cells; see experiment 2.
Experiment 2
In a first experiment were sowed colon car-
cinoma cells under the influence of "EFLA 85942" in so
called adhered culture flasks.
Expected was the formation of a so called mo-
nolayer-film.
But quite surprisingly was observed the forma-
tion of three-dimensional, strongly connected cell ag-
gregates.
Also this behaviour has not yet been observed.
In addition were observed in the culture media
nearly no non-adhered single cells.

CA 02217227 1997-10-24
In a second experiment some few single cells
were sowed in non-adhered Petri dishes, with the aim to
analyze the influence of "EFLA 85942" onto the colony
generation ability of these single cells.
Thereby two variants were carried out.
In experiment A was added "EFLA 85942" to the
cells after an initial adhesion on the Petri dish.
In experiment B was added "EFLA 85942" to the
cells immediately before the adhesion process.
In these two experiments could be determined a
statistical significant decrease of the colony generati-
on ability which occured in experiment B, in comparision
to experiment A, in a drastic stronger extent.
The result of these two experiments, together
with the above described formation of three-dimensional
cell aggregates, allows the conclusion that on one hand
the adhesion with non histo-compatible structures is re-
duced and that on the other hand the adhesion betweeen
single, to the same tissue type belonging cells or cell
unions is promoted.
These behaviour characteristics prove that be-
side the above mentioned antioxidative effect of the po-
lyphenols still further effect mechanisms and/or further
active compounds play an important rule.
The treatment of the human or animal body may
consist therein that the corresponding preparation is
taken or is applied internal or external.

CA 02217227 1997-10-24
The treatment is carried out at least during
such a long time until the corresponding symptomatic has
disappeared.
The following examples illustrate the present
S invention.
Example 1
Preparing of a preparation in the form of a
hard gelatin capsule.
For the preparation of 1000 capsules of the
size "one " 100 g of "EFLA 85942" were mixed homogene-
ously with 25 g of microcristalline cellulose, 4 g of
magnesium stearate and 1 g of precipitated silicic acid.
This mixture was filled in a respective amount
of 150 mg into hard gelatin capsules.
Example 2
Preparing of a preparation in the form of a
drinking ampule.
For the preparation of 1000 drinking ampuls of
10 ml 100 g of "EFLA 85942" were mixed homogeneously
with 15 g of potassium sorbate, 15 g of sodium benzoate,
500 g of fructose and 10 g of sodium chloride.
This mixture was filled up with a solution of
95 parts by weight of water and 5 parts by weight of
glycerol to a total volume of 10 liters, mixed and fil-
tered sterile.

CA 02217227 1997-10-24
The obtained filtrate was filled aseptic into
the drinking ampules.
Example 3
Preparing of a preparation in the form of a
liposome gel.
Phase A
- 1.0 parts by weight of RhodigelR 200,
- 8.0 parts by weight of 1,2-propylene glycol
were mixed at room temperature and were allo-
wed to swell during one hour.
Phase B
- 6.5 parts by weight Phospholipon 80,
- 10.0 parts by weight of polyethylene glycol
400,
- 2.0 parts by weight of glycerol
were mixed together at a temperature from 60~C
to 70~C and were homogenized.
The obtained homogeneous mixture was cooled to
a temperature from 38~C to 40~C under stirring.
Phase C
A mixture of

CA 02217227 1997-10-24
- 71.7 parts by weight of distilled water,
- 1.2 parts by weight of "EFLA 85942"
was filtered sterile and was then incorporated
drop by drop into the phase B. After the complete addi-
tion the obtained mixture was cooled to room tempera-
ture, incorporated in portions into the phase A and ho-
mogenlzed.
The obtained preparation can be filled in pla-
stic squezze bottles of 10 ml.
The preparation was prepared unter aseptic
conditions.
Application example 1
A 30 years old female healthy test person with
head hair problems in the form of thin areas and par-
tially growed places took over a time of 8 weeks daily 3capsules according to example 1.
Within this time the state of the head hair
changed drastically.
In the growed hair areas germinated normally
coloured hairs, and the thin areas disappeared.
Application example 2
A 36 years old male healthy test person with
head hair problems in the form of thin areas and par-
tially growed places took over a time of 8 weeks daily 6
capsules according to example 1.

CA 02217227 1997-10-24
Within this time the state of the head hair
changed drastically.
In the growed hair areas germinated normally
coloured hairs, and the thin areas disappeared.