Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Conjugate or complex
Reexamination Certificate
2000-04-17
2001-08-14
Lankford, Jr., Leon B. (Department: 1651)
Drug, bio-affecting and body treating compositions
Antigen, epitope, or other immunospecific immunoeffector
Conjugate or complex
C530S395000, C514S931000, C514S934000
Reexamination Certificate
active
06274146
ABSTRACT:
BACKGROUND OF THE INVENTION
The invention refers to biologically active substances and, in particular, to phytogenous proteoglycans applied in medicine, pharmacology, veterinary medicine, and biology as a component of medicinal products.
Yeast glucan is known that increases resistance in experimental animals to bacterial, fungal, viral, and parasitic infections. For example, Browder et al. have shown in Int. Immunopharm, 1984, 6, #1 pp. 19-26, that the administering of yeast glucan before intravenous contamination of
Staphilococcus aureus
mice resulted in almost double increase in infected animal lifetime. On a model of gram-negative bacterial sepsis the peritoneal injection of yeasty glucan considerably reduced system bacteremia and increased animals survival rate as was shown by D. L. Williams, et al (J. Reticuloendothelial Soc., 1978, V. 23, pp. 479-490). The same authors demonstrated the efficiency of glucan with viral hepatitis in mice. Glucan administration supported Kupfer's cell phagocyte activity, promoting thereby regeneration of hepatocytes However, with application of yeast glucan the formation of granulomas in liver and the development of allergic diseases was observed.
Bioactive polysaccharide &ggr;-PL (gamma-plant) is also known, which is extracted from plant cells, for example from corn, potato, marine fungus, etc. With the molecular mass equal to 2×10
6
±9×10
5
D and element composition (mass %) as follows: nitrogen 1.7-1.98; carbon 40.12-40.39; hydrogen 5.81-6.07; the remainder is the ash component which includes the polysaccharide chain consisting of (mass %) neutral carbohydrate residues (35.0-41.0), glucose (27.0-33.0), galacturonic acid (19.0-25.0), arabinose (1.7-2.3), uronic acids (12.0-18.0) and protein (no less than 0.5) that consists of amino acids residues in the following quantities (per 0.1 g of ⊖-pl): asparagin 126.0-146.0, serine 139.0-159.0, glutamine 263.0-283.0, glycine 117.0-131.0, alanine 80.0-100.0, valine 76.0-96.0, leucine 85.0-105.0, lysine 65.0-85.0, arginine 42.0-62.0; in trace quantities are: cystein, isoleucine, histidine, phenylalanine, tyrosine, threonine. As an anti-infective agent gPL was studied during treatment of viral and bacterial infections in laboratory and agricultural animals. With experimental infection of mice by viruses of simple herpes such as 1 and 2 types on the model of herpetic meningocephalitis at application of a dose 10LD
50
gPL protective effect reached 60%. Comparatively narrow range of anti-infective activity as well as an occurrences of local inflammatory reactions while subcutaneous injections are gPL applications shortcomings.
SUMMARY OF THE INVENTION
It is therefore an object of the present invention to eliminate the shortcomings mentioned above by creation of a new substance having a broad spectrum of anti-infectious activities and free from side effects.
In keeping with these objects and with others which will become apparent hereinafter, one feature of the present invention is to provide a substance which is the gPL—new biologically active phytogenous proteoglycan obtained by disintegration of divisible plant cells, for example, from young potato (
Solanum tuberosum,
family Solanacae) plants via water solution. More particularly, in accordance with the present invention, a new biologically active phytogenous proteoglycan is provided, which includes a biologically active phytogenous proteoglycan and not developing hemagglutinating activity and obtained by disintegration of divisible plant cells, for example, from young potato (
Solanum tuberosum
, family Solanancae) plants via water solution, fractionated and concentrated to obtain a substance with a molecular mass of 8.0×10
5
-
2
.
5
×
10
6
D and the following composition (mass %): nitrogen 1.12-2.48; carbon 39.93-44.42; hydrogen 5.15-7.21; the remaining is the ash component which includes the polysaceharide chain consisting of (mass %) residues of neutral sugars (34.0-85.3), glucose (26.4-33.1), galacturonic acid (19.0-25.1), arabinose (1.7-4.4), uronic acids (12.0-18.0), ramnose (1.2-10.0), xylose (0.1-3.0), mannose (0.1-5.0), galactose (2.5-27.0), and protein (up to 15.0) that consists of amino acids residues in the following quantities (ng per 0.1 mg of proteoglycan): asparagin 126.0-146.0, serine 139.0-159.0, glutamine 263.0-283.0, glycine 117.0-131.0, alanine 80.0-100.0, valine 76.0-96.0, leucine 85.0-105.0, lysine 65.0-85.0, arginine 42.0-62.0; in trace quantities there are: cystein, isoleucine, histidine, phenylalanine, tyrosine, threonine.
In accordance with another feature of present invention, a method for producing a biologically active proteoglycan which is proposed which includes the steps of shredding into mush-like condition a raw material in from germinated potato bulbs
Solanum tuberosum,
Solanacae family extracting with boiling water (with water: material rate 1.1:1.2), holding for 23÷25 h at 20° C., separating by filtration of an extract obtained into a solid and a liquid phase, fractionating the latter by means of either gel-chromatography or filtration to remove fractions with a molecular weight not exceeding 8.0×10
5
D, concentrating the extract obtained to an end-product in form of a dry powder.
The novel features which are considered as characteristic for the present invention are set forth in particular in the appended claims.
DESCRIPTION OF PREFERRED EMBODIMENTS
In accordance with the present invention, a substance is proposed which is the gPL—new biologically active phytogenous proteoglycan obtained by disintegration of divisible plant cells, for example, from young potato (
Solanum tuberosum,
family Solanacae) plants via water solution fractionated in and concentrated to obtain a dry substance with a molecular mass 8.0×10
5
-2.5×10
6
D, and with composition (mass %) is as follows: nitrogen 1.12-2.48; carbon 39.93-44.42; hydrogen 5.15-7.21; the remaining is the ash component which includes the polysaccharide chain consisting of (mass %) residues of neutral sugars (34.0-85.3), glucose (26.4-33.1), galacturonic acid (19.0-25.1), arabinose (1.7-4.4), uronic acids (12.0-18.0), ramnose (1.2-10.0), xylose (0.1-3.0), mannose (0.1-5.0), galactose (2.5-27.0), and protein (up to 15.0) that consists of amino acids residues in the following quantities (ng per 0.1 mg of proteoglycan): asparagin 126.0-146.0, serine 139.0-159.0, glutamine 263.0-283.0, glycine 117.0-131.0, alanine 80.0-100.0, valine 76.0-96.0, leucine 85.0-105.0, lysine 65.0-85.0, arginine 42.0-62.0; a trace quantities that are: cystein, isoleucine, histidine, phenylalanine, tyrosine, threonine. Proteoglycan is characterized by the following infra-red spectrum peaks (KBr tablet): 3350, 2920, 1270, 1655, 1620, 1610, 1560, 1510, 1405, 1378, 1296, 1145, 1080, 1040, 920, 842, 763 cm
−1
. The substance in question is insoluble in organic solvents and soluble in aqueous and saline solvents, possesses anti-infectious properties and does not possess hemagglutinating properties.
Proteoglycan element composition is distinct from that of gamma-plant due to the fact that, apart from monosaccharides, which are present in gamma-plant, it contains ramnose, xylose, mannose, and galactose and has a wide anti-infectious spectrum.
A method of producing the new proteoglycan is described hereinbelow.
The method involves raw material shredding, extraction by water, keeping at the temperature of 20° C. within 24 hours, extract separation and, finally, concentrating to a dry substance. In accordance with the invention, young root plants of potato
Solanum tuberosum
(Solanaseae family) bulbs are used as raw material. The process of extraction is carried out with boiling water or water having PH value in the limits of 4,5-6,5. The extract obtained is fractionated and concentrated to molecular mass to remove low molecule fractions with the weight not exceeding 8.0×10
5
D.
The method will be realized in the following way: roots that growing out from eyelets in potato bulbs are used as a raw material. Thus, th
Flood Michele
Lankford , Jr. Leon B.
Zborovsky I..
LandOfFree
Biologically active phytogenous proteoglycan and a method of... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Biologically active phytogenous proteoglycan and a method of..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Biologically active phytogenous proteoglycan and a method of... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2541429