WO2002017934A1

WO2002017934A1 - Natural, non-allergenic, immune system stimulant

Info

Publication number: WO2002017934A1
Application number: PCT/US2000/024428
Authority: WO
Inventors: Paul Yau Sun Lam
Original assignee: ASIA CAPITAL GROUP A BRITISH VIRGIN ISLANDS Corp % CHINESE HERBAL USA Inc
Current assignee: ASIA CAPITAL GROUP A BRITISH VIRGIN ISLANDS Corp % CHINESE HERBAL USA Inc
Priority date: 2000-08-31
Filing date: 2000-08-31
Publication date: 2002-03-07
Anticipated expiration: 2003-02-28
Also published as: AU2000271165A1

Abstract

A mixture of 5-80 % by weight Hericium, 5-40 % by weight Radix Astragali, 5-40 % by weight Cordyceps Sinensis, 1-40 % by weight Radix Glycyrrhizae and 5-40 % by weight Dioscoreae Oppositae. Testing shows that this mixture produces an increase in IL1 beta without an increase in 1L4.

Description

Natural. Non-Allerαenic. Immune System Stimulant

Technical Field

This invention relates to herbal dietary supplements. More particularly this invention relates to supplements which will stimulate the immune system without producing an allergic response.

Background Art

Recently, evidence has been accumulating that herbal supplements, taken regularly, may have beneficial effects. Herbal supplements, unlike medicines are not effective instantaneously but must be taken over long periods of time in order be effective. In other word, it takes time to create an effect in humans. There are many products on the market that claim to stimulate the immune system but the research behind such products seems to be borderline at best. See article on Functional food to be published in California Agriculture 2000, probably in November 2000 issue. Stimulating the immune system is a perfect application for herbal supplements because the immune system can be improved gradually. An improved immune system can be beneficial in fighting all kinds of illnesses from the common cold to deadly cancers.

Unfortunately, many herbal supplements can create an allergic response in humans. What is needed is a dietary supplement or herbal supplement that can stimulate the human immune system without allergic response. It is also important that this product have been properly tested for efficacy. Development of an herbal supplement which can stimulate the immune system without producing an allergic response represents a great improvement in the field of medicine and satisfies a long felt need of medical practitioners and their patients. Disclosure of Invention

This invention is a mixture of five natural herbs. This invention comprises: 5- 80% by weight Hericium, 5-40% by weight Radix Astragali, 5-40% by weight Cordyceps Sinensis, 1-40 % by weight Radix Glycyrrhizae and 5-40% by weight Dioscoreae Oppositae. Preferably the powders are size 80 i.e. the powders will pass through a sieve with 80 holes. This is a Chinese standard used in measuring herbal powder.

The mixture can be administered in any acceptable form such as in capsules, pressed into tablets, and made into gel caps. Gel caps are a recent development in which a powder is placed in suspension in a fluid and then is encapsulated with a gel. The preferred dosage is 100mg of the invention, twice a day, 100 mg. at breakfast and 100 mg. before bed.

This invention was tested by the following procedure at the Department of Clinical Immunology at the University of California at Davis:

I. Collect blood.

2. Isolate white cells.

3. Isolate lymphocytes

4. Incubate lymphyocytes in mini tubes for varying periods in the presence or absence of the invention. 5. Incubate the lymphocytes with 4 serial ten fold dilutions of the invention from 0.1 to 100 μg/ml

6. Repeat the same experiments in the presence or absence of phytohemagglutinin (PHA), a known stimulator of cells

7. Collect the culture supernatants (i.e. the fluid above the lymphocytes). 8. Using known standards and a reference curve assay supernatants for IL1¹ beta and for IL4.

^l\L is the standard abbreviation for interleukin. The results showed an impressive production of IL1 beta without an increase in 1 L4. Such production would be helpful during periods of needed immune response, i.e. during a cold or infection.

An appreciation of the other aims and objectives of the present invention and a more complete and comprehensive understanding of it may be achieved by studying the following description of the best mode of carrying out the invention.

Best Mode for Carrying Out Invention

This invention comprises the following mixture. All ingredients are naturally occurring herbs.

Latin Name Weight %

Hericium 5 - 80

Radix Astragali 5 - 40

Cordyceps Sinensis 5 - 40

Radix Glycyrrhizae 1 - 40

Dioscoreae Oppositae 5 - 40

Example: In Vitro Analysis of Herbal Product

Treatment Preparation. Ten mg of herbal product was suspended in 1 ml RPMl²-1640 with 10% heat inactivated fetal bovine serum (media). The suspension was shaken vigorously on a vortex mixer. The insoluble components were quickly pelleted and the remaining suspension filtered twice: first through a 0.45 micron syringe filter followed by a 0.2 micron syringe filter. This filtrate was used as the stock (approx. 10 mg/ml) from which dilutions were made for the

This is a neutral, buffered, solution containing nutrients and special salts, which is designed for growing cells. RPMI stands for Roswell Park Memorial Institute. incubations with peripheral blood mononuclear cells (PBMC). The stock solution was diluted in media prior to treatment with PBMC.

PBMC Isolation. Peripheral blood from six healthy volunteers was collected and the PBMC were isolated over a density gradient via centrifugation. The cells were re-suspended in RPMI-1640 containing 10% fetal bovine serum and supplemented with 0.1% of a 50 mg/ml gentamicin solution (Gibco BRL, Grand Island Nebraska). PBMC concentration was adjusted to 2 x 106 viable cells/ml after estimation of viability by trypan blue exclusion assay. Viability was consistently greater that 96%.

Culture of PBMC with Herbal Products. Five hundred ml of a 1.0 x 106 cell suspension were cultured with equal volume of the Product treatments at 37°C with 5% C0₂ in 48-well plates. PBMCs were exposed to a final concentration of 100, 10, 1 , 0.1 mg/ml of product. In addition, each product treatment was incubated in the presence of PHA at 10 mg/ml. Culture supernatant fractions were harvested after 72 h and were stored at -20°C until analysis by enzyme linked immunoassay (ELISA).

ELISA Analysis of Cytokines. Levels of IL1 , IL4, and gamma interferon (IFN) were measured in supernatants from 1.0 x 106 cells/ml stimulated with Product in the presence or absence of PHA. For quantitation of secretory IL4, a High Sensitivity Quantikine Human IL4 ELISA kit was used with a detection limit of 0.25 pg/ml (R&D Systems, Minneapolis, MN). Standard ELISA kits were use to quantitate IL1 and IFN (R&D Systems, Minneapolis, MN) with detection limits of 3.9 and 31.2 pg/ml respectively.

Results showed an increase of IL1 beta of between 6 and 11 times without any increase in IL4. In other words this product stimulates the human immune system without stimulating IL4, a precursor to allergies in the human body. This testing was conducted by M. Eric Gershwin M.D., The Jack and Donald Chia Professor of Medicine, Chief, Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis.

This invention has been described with reference to a particular embodiment. However, it should be obvious to those skilled in the art to which this invention pertains that other modifications and enhancements can be made without departing from the spirit and scope of the claims that follow.

Claims

1. An immune system stimulator comprising 5-80% by weight Hericium, 5-40% by weight Radix Astragali, 5-40% by weight Cordyceps Sinensis, 1-40 % by weight Radix Glycyrrhizae and 5-40% by weight Dioscoreae Oppositae.

^' 5 2. An immune system stimulator as claimed in claim 1 in which all ingredients are powdered.

3. An immune system stimulator containing a biologically effective amount of the immune system stimulator claimed in claim 1.

10

4. An immune system stimulator as claimed in claim 3 in which said biologically effective amount is 100 mg.

5. An immune system stimulator as claimed in either of claim 1 , 2 or 4 in which 15 said ingredients are incorporated into a pill.

6. An immune system stimulator as claimed in either of claim 1 , 2 or 4 in which said ingredients are contained in a capsule.

20 7. An immune system stimulator as claimed in either of claim 1 , 2 or 4 in which said ingredients are contained in a gel cap.

8. A method of making an immune system stimulator comprising the steps of: a. obtaining powdered Hericium, Radix Astragali, Cordyceps Sinensis, 25 Radix Glycyrrhizae and Dioscoreae Oppositae; and b. mixing 5-80% by weight Hericium, 5-40% by weight Radix Astragali, 5- 40% by weight Cordyceps Sinensis, 1-40 % by weight Radix Glycyrrhizae and 5-40% by weight Dioscoreae Oppositae together.

9. A method of making an immune system stimulator as claimed in claim 8 further comprising the step of forming said mixture into a pill.

10. A method of making an immune system stimulator as claimed in claim 9 in which said pill contains 100 mg. of said mixture.

11. A method of making an immune system stimulator as claimed in claim 8 further comprising the step of incorporating said mixture within a capsule.

12. A method of making an immune system stimulator as claimed in claim 9 in which said capsule contains 100 mg. of said mixture.

13. A method of making an immune system stimulator as claimed in claim 8 further comprising the step of incorporating said mixture within a gel cap.

14. A method of making an immune system stimulator as claimed in claim 9 in which said gel cap contains 100 mg. of said mixture.