Drug – bio-affecting and body treating compositions – Plant material or plant extract of undetermined constitution... – Containing or obtained from a leaf
Reexamination Certificate
2000-12-26
2003-03-11
Tate, Christopher R. (Department: 1651)
Drug, bio-affecting and body treating compositions
Plant material or plant extract of undetermined constitution...
Containing or obtained from a leaf
C424S725000, C424S727000, C514S885000
Reexamination Certificate
active
06531166
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to use of betel leaf extract to induce IFN-gamma production from human peripheral blood T cells.
BACKGROUND AND PRIOR ART REFERENCES
Stinging nettle leaf extracts are registered in Germany for adjuvant therapy of rheumatic diseases. In a whole blood culture system, the nettle extract IDS 23 (Rheuma-Hek) inhibited lipopolysaccharide stimulated monocyte cytokine expression, indicating an immunomodulating effect. (Antirheumatic Effect of IDS 23, a Stinging Nettle Leaf Extract, on in vitro expression of T helper cytokines). The applicants investigated the immunomodulating effects of betel leaf extracts on phytohemagglutinin (PHA) stimulated peripheral blood mononuclear cells (PBMC) in vitro. Betel leaves has a strong pungent aromatic flavor and widely used as a masticatory. Generally, mature or over mature leaves, which has ceased growing but not yet become brittle are used for chewing. The basic preparation for chewing purposes consists of betel leaf smeared with hydrated lime and catechu to which scrapings of arecanut are added; flavorings such as coconut shavings, clove, cardamon, fennel, powdered liquorice, nutmeg and also tobacco are used according to one's taste. In some places prepared pan is covered with silver or gold leaf. As a masticatory, it is credited with many properties: it is aromatic, digestive, stimulant and carminative. Medicinally it is useful in catarrhal and pulmonary affections; it is also used for poultices. The effects of chewing of betel with arecanut and other adjuncts are the excitation of the salivary glands and the irritation of the mucous membrane of the mouth. The red coloration produced is due to a pigment in the arecanut, which manifests itself under the action of alkali in lime and catechu. A mild degree of stimulation is produced, resulting in a sensation of warmth and well being, besides imparting a pleasant odor. The most important factor determining the aromatic value of the leaf is the amount and particularly the nature of the essential oil present. Betel leaves from different regions vary in smell and taste. The most pungent is the Sanchi type, while the most mild and sweet ones are from Madras. The betel leaves contain essential oils, the content of oil varies from 0.7 to 2.6 per cent depending upon the varieties of leaves. The oil consists of phenols and terpens. The higher the proportion of phenol oil, the better the quality. An isomer of eugenol named chavibetol (betel phenol; 4-allyl-2-hydroxy-1-methoxy benzene) is considered to be the characteristic constituent of betel oil. It is however, absent in Indian samples. Betel oil of Indian types contain as a predominant phenolic constituent. Oil of betel has been used in the treatment of various respiratory catarrhs, under as a local application either by gargle or by inhalation in diphtheria. It has carminative properties. It exhibits in different action on the central nervous system of mammals; lethal doses produce deep narcosis leading to death within a few hours. The essential oil and extracts of the leaves possess activity against several Gram-positive and Gram-negative bacteria such as
Micrococcus pyogenes
var.
albus
and var.
aureus, Bacillus subtilis
and
B. megaterium, Diplococcus pneumoniae, Streptococcus pyogenes, Escherichia coli, Salmonella typhosa, Vibrio comma, Shigella dysenteriae, Proteus vulgaris, Pseudomonas solanacaerum, Sarcina lutea
and
Erwinia carotovora
. The oil is found to be lethal in about 5 minutes to the protozoa
Paramaecium caudatum
(Wealth of India, Vol.8 pp.84-94). It inhibits the growth of
Vibrio cholerae, Salmonella typhosum
and
Shigella flexneri
and
Escherichia coli
. Steam—distillate of the leaves showed activity against
Mycobacterium tuberculosis.
OBJECTS OF THE INVENTION
The main object of the present invention is to provide a method for inducing IFN-&ggr; from human peripheral blood mononuclear cells.
Another object of the present invention relates to use of betel leaf extract for inducing IFN-&ggr; production from human peripheral blood T cells.
Another object of the invention is to provide a composition comprising betel leaf extract, which is useful as Th
1
type immunomodulator.
SUMMARY OF THE INVENTION
To meet the above objects, the present invention provides a method for inducing IFN-&ggr; from human peripheral blood mononuclear cells. The invention also relates to use of betel leaf extract for inducing IFN-&ggr; production from human peripheral blood T cells.
DETAILED DESCRIPTION OF THE INVENTION
Accordingly, the invention relates to a method for inducing IFN&ggr; from human peripheral blood mononuclear cells wherein the said method comprising preparing water extract of betel leaf; preparation of human peripheral blood mononuclear cells; incubation of hPBMC with betel leaf extract for a period of 18-48 hours; extraction of RNA for cytokine specific RT-PCR or for flow cytometry for the detection of intracellular cytokine protein; subjecting RNA for RT-PCR to obtain PCR products using IFN&ggr; specific known primers and enhancement of IFN&ggr; as reflected by IFN&ggr; specific band.
Alternatively, one more method for inducing IFN&ggr; produced from human peripheral blood mononuclear cells wherein the said process comprising subjecting incubated cells for intracellular staining for IFN&ggr;; analysis of stained cells in flow cytometer; and enhancement of IFN&ggr; positive cells to at least seven fold.
A method for using betel leaf extract as Th
1
type immuno-modulator wherein the said method comprising:
a) administering to a subject at least 5 to 10 mg/ml/kg body wt. of betel leaf extract, and
b) administering the extract through oral or intramuscular route once in a day for a period of at least one month,
In an embodiment of the present invention provides a pharmaceutical composition useful as Th
1
type immunomodulator, said composition comprising an effective amount of betel leaf aqueous extract or lyophilized betel leaf extract together with or associated with an additive.
In still another embodiment of the invention, the additive is selected in such a manner that does not interfere with the activity of betel leaf extract.
Yet another embodiment of the invention, the additive is selected from nutrients such as proteins, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gelatin paste and/or pharmaceutically acceptable carriers, excipient, diluent or solvent.
Yet another embodiment of the invention, the aqueous extract, lyophilized product or the composition is administered orally or intramuscularly.
Yet another embodiment of the invention, the oral route is in the form of capsule, syrup, concentrate, powder or granules.
Yet another embodiment of the invention, the ratio of betel leaf extract to the additive is in the range between 10-1:1-10.
Yet another embodiment of the invention, the betel leaf extract, lyophilized extract or the composition comprising the betel leaf extract is administered at a dosage level between 5 to 10 mg/kg of body weight at least once in a day for one month.
In one more embodiment of the present invention, a method of treating a subject to provide Th1 type immuno-mudulation, said method comprising administering a pharmaceutically effective amount of betel leaf extract, lyophilized extract or a composition comprising the extract to the subject.
Yet another embodiment of the present invention, the additive is selected in such a manner it does not interfere with the activity of lyophilized betel leaf extract.
Yet another embodiment of the invention, the additive is selected from nutrients such as proteins, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gelatin paste and/or pharmaceutically acceptable carriers, excipient, diluent or solvent.
Yet another embodiment of the invention, the aqueous extract, lyophilized product or the composition is administered orally or intramuscularly.
Yet another embodiment of the invention, the oral route is in the form of capsule, syrup, concentrate
Bandyopadhyay Santu
Bhattacharya Samir
Chandra Roy Keshab
Pal Bikash
Ray Mitali
Council of Scientific and Industrial Research
Tate Christopher R.
LandOfFree
Use of betel leaf extract to induce IFN-gamma production... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Use of betel leaf extract to induce IFN-gamma production..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Use of betel leaf extract to induce IFN-gamma production... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3021147