Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Carbohydrate doai
Reexamination Certificate
1999-06-02
2002-11-26
Priebe, Scott D. (Department: 1632)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Carbohydrate doai
C536S023100, C424S234100, C424S241100, C424S245100, C424S246100
Reexamination Certificate
active
06486132
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to an immunomodulator, an immunomodulator food, an immunomodulator feed, a troche and an agent for external application, having DNA derived from the cells of procaryotes.
2. Related Art
Appropriate activation of an immunological activity of man and animals is considered important to become resistant to infectious diseases including common cold, to primary prevention of cancers, and checking allergic and atopic diseases and the like. It has been known that DNA from the cells of procaryotes has an immunopotentiation activity and DNA from the cells of eucaryotes does not. However, the immunomodulation activity of DNA is inactivated through deoxyribonuclease treatment in vitro (S. Yamamoto et al., Microbiol. Immunol., Vol. 36(9), 983-997, 1992). It has been so far considered that since a digestive juice enzyme of humans and animals contains deoxyribonuclease, the immunomodulation activity of DNA is effective only through administration thereof by injection.
Since the administration by injection damages the body, however, it is inappropriate for repeated administration. In order to improve the physical condition for which immunopotentiation is deemed necessary, it is actually rather important that the purpose is achieved by daily intake of DNA through food or by contact with the cutaneous mucosa.
SUMMARY OF THE INVENTION
Accordingly, an object of the present invention is to provide an immunomodulator in which DNA can be administered not through injection, but, for example, orally or percutaneously, or can daily be taken in food or feed.
As a result of extensive investigation in order to solve the above-mentioned problem, the present inventors have found that the exhibition of immunomodulation activity is unexpectedly obtained by either oral or percutaneous administration of DNA derived from procaryotes such as
Bacillus subtilis
and lactic acid bacteria. This finding has led to the completion of the present invention.
That is, the present invention is to provide an immunomodulator, an immunomodulator food or an immunomodulator feed containing DNA derived from cells of procaryotes as an active ingredient.
The invention provides a method of modulating the immunofunction by administering orally, percutaneously or permucosally a pharmacologically effective amount of DNA extracted from cells of a procaryote to human being or animal.
It is preferable that the DNA is a product produced by culturing cells of a procaryote, harvesting the cells, dissolving the cells with a bacterial cell wall digesting enzyme in water and adding ethanol to the lysate solution to isolate a DNA product. The DNA may be the DNA extraction product or the ethanol-insoluble fraction in ethanol or an aqueous ethanol of bacterial cells. The product may be well obtained at about 40% volume/volume aqueous ethanol.
It is preferable that the DNA has at least 8 bases and includes at least one CpG sequence. The DNA a sodium salt of the same.
In the invention, the reticuloendothelial system is activated. Accordingly, an infectious disease can be treated.
In the invention, the growth of a tumor is inhibited.
In the invention, an autoimmune cutaneous disease is treated.
DETAILED DESCRIPTION OF THE INVENTION
In the present invention, the procaryotes are not necessarily specific bacteria. However, in view of the safety,
Bacillus subtilis
(genus Bacillus), lactic acid bacteria (lactobacilli), amino acid-producing bacteria,
Escherichia coli
and the like can be mentioned. Of these,
Bacillus natto,
lactic acid bacteria and amino acid producing bacteria, which are used in production of foods, are especially preferable. As lactic acid bacteria, preferably,
Lactobacillis casei, Lactobacillus acidophilus, Lactobacillus del
-
brueckii, Streptococcus faecalis, Biphidobacterium longum
and the like can be mentioned. As amino acid producing bacteria, for example,
Corynebacterium glutamicum, Corynebacterium herculis, Brevibacterium flavum, Microbacterium ammoniaphilum
and
Escherichia coli
can be mentioned.
DNA can be removed from cells of procaryotes by a known method. For example,
Corynebacterium glutamicum
is cultured in an appropriate medium. The cells are then harvested through centrifugation, washed with a 0.85% physiological saline solution, and thereafter suspended in a physiological saline solution or purified water. The cells are dissolved with the addition of a bacterial cell wall digesting enzyme such as a lysozyme. To this solution is gradually added pure ethanol that has been cooled. DNA which is insolubilized at an ethanol concentration of 40% volume/volume is wound up on a glass bar. The resulting DNA is washed with 60% volume/volume ethanol, with 70% volume/volume ethanol and with 80% volume/volume ethanol in this order, and dried in vacuo to obtain a high-purity DNA product.
The resulting DNA product can be added as such to food, feed or beverages. It is also possible that the resulting DNA product is mixed with a filler, an excipient and the like which are ordinarily the production of pharmaceuticals, and the mixture is molded into an appropriate form as required to provide preparations or food. It can further be formulated into a solution.
The immunomodulation activity of the procaryotic DNA is chemically stable. Even when it was added to an ordinary feed, it neither reacted with feed ingredients nor deactivated with heat as described in Examples 4 and 5. This shows that during the step of producing the immunomodulator food and the immunomodulator containing this DNA, cooking or heat sterilization treatment can freely be practiced. The immunomodulation activity of the DNA derived from procaryotes is, as mentioned above, stable, and no aging deterioration is observed. Accordingly, the activity is not lost during the storage or the distribution of this product.
In case of administering DNA as an immunomodulator in the form of an aqueous solution, it is especially rendered to have a low-molecular weight through ultrasonic treatment (sonication) or restriction endonuclease treatment, whereby the viscosity decreases and it becomes convenient for drinking. In the sonication, for example, purified water is added to the DNA product such that the concentration reaches 1%, and an ultrasonic wave is applied thereto for from 5 minutes to 20 minutes while cooling the same with ice water. It has been known that the DNA molecule derived from procaryotic cells exhibits immunomodulation activity because the base sequence is characteristic and procaryotic cells, unlike eucaryotic cells, contain a large number of CpG sequences (A. M. Krieg et al., Nature, Vol. 374; pp. 546-549, 1995). Accordingly, in the restriction endonuclease treatment, the restriction endonuclease is not particularly selected so long as it does not cleave a CpG linkage. The restriction endonuclease treatment may be conducted by an ordinary endonuclease treatment method.
The food is not particularly limited when DNA is provided in food through addition. DNA can be added to, for example, staple foods such as bread, noodles and rice; side dishes such as boiled fish paste (kamaboko), ham, sausage, cooked salad and pickled vegetables; seasonings such as soy-bean paste (miso), soy sauce, soup, sauce and dressing; dairy products such as milk, cocoa drinks, butter, margarine, cheese and ice cream; confectioneries such as cookies, biscuits, wafers, candies and chewing gum; processed fruit products such as fruit juice drinks and jam; and pet feed, fishery and stockbreeding feeds and the like.
When DNA in the present invention is applied as a main agent, it can orally be used in powder, granules, dispersions, capsules, sweets, drinks or the like. In addition, it can be used percutaneously as an ointment, a compress, a solution or a bath product and permucosally as a troche, a suppository or a gargle. In the percutaneous use, the above-mentioned low-molecular product is preferable. When the immunomodulator of the present invention is used through oral administration, the standard inta
Fukuda Yasuki
Katsuyama Hiroichi
Murata Kousaku
Yajima Mizuo
Asama Chemical Co., Ltd.
Chen Shin-Lin
Flynn ,Thiel, Boutell & Tanis, P.C.
Priebe Scott D.
LandOfFree
Immunomodulator, immunomodulator food and immunomodulator feed does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Immunomodulator, immunomodulator food and immunomodulator feed, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Immunomodulator, immunomodulator food and immunomodulator feed will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2958023