Acidic polysaccharide CH-1 isolated from Chlorella pyrenoidosa and the use thereof

• US 4,533,548 A
• Filed: 11/15/1982
• Issued: 08/06/1985
• Est. Priority Date: 12/02/1981
• Status: Expired due to Fees

• Current Assignee: Kitasato Institute (Kitasato University)
• Original Assignee: Kitasato Institute (Kitasato University)
• Inventors: Komiyama, Kanki, Umezawa, Iwao
• Primary Examiner(s): Wiseman, Thomas
• Assistant Examiner(s): Teskin, Robin
• Application Number:
  US06/441,630
• Time in Patent Office: 995 Days
• Field of Search: 424/195, 424/180, 424/88, 435/946, 435/101, 260/112 R, 536/1
• US Class Current: 514/54
• CPC Class Codes: A61K 31/715 Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
• CPC Class Codes: C12N 1/125 Unicellular algae isolates
• CPC Class Codes: C12P 1/00 Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
• CPC Class Codes: C12P 19/26 Preparation of nitrogen-containing carbohydrates
• CPC Class Codes: C12R 2001/89 Algae ; Processes using algae
• CPC Class Codes: Y10S 435/946 using algae

Abstract

A new acidic polysaccharide is isolated from a culture of cells of the algae Chlorella pyrenoidosa. The process for producing this polysaccharide is also disclosed. This substance exhibits antitumor activity and antiviral activity and induces the production of interferon.

Claims

1. An acidic polysaccharide having the following properties:

(a) consisting of carbon, hydrogen, oxygen and nitrogen with an approximate elementary analysis of O;

54.05%;

C;

38.49%;

H;

6.07%; and

N;

1.39%;

(b) a sedimentation coefficient of S₂₀,w=6.15 S when determined by ultracentrifugation;

(c) impermeability to cellophane membrane,

(d) a melting point characterized by coloration at about 240°

C. with carbonization at higher temperatures;

(e) a specific rotation of [α

]_D²⁹ =-9.52 (c=1, H₂ O);

(f) solubility in water, and insolubility in methanol;

(g) a positive color reaction with anthrone, Molisch, and carbazole-H₂ SO₄ and a negative color reaction with Folin-Lowry and ninhydrin;

(h) no detectable protein as measured by Folin-Lowry color reaction test;

(i) a monosaccharide analysis consisting of rhamnose, galactose, arabinose, glucose and glucuronic acid when detected by paper chromatography and gas chromatography after hydrolysis of said polysaccharide in 7% H₂ SO₄ under refluxing for two hours;

(j) a UV spectrum with end absorption in 0.125% aqueous solution;

(k) an IR spectrum (KBr tablet) with absorption bands at 3420, 2930, 1640, 1380, 1125, 1065, 1040, 990, 915, 835 and 880 cm^-1 ;

(l) an electrophoretic pattern on acetate membrane as shown in FIG. 2, electrophoresced to 0.8 cm for (+) with 0.1M pyridine-acetic acid at pH 5.0, 200 V (4 mA) for 20 minutes;

(m) an ultra-centrifuge pattern as shown in FIG. 3;

(n) being obtained as a white powdery precipitate; and

(o) having a sugar content of 53% when calculated as glucose.

2. A method for the production of an acidic polysaccharide of claim 1, comprising extracting spray-dried powder of Chlorella pyrenoidosa with hot water at about 80°

-90°

C. and fractionally precipitating the polysaccharide with a water-miscible C₁ -C₄ alkanol in which it is substantially insoluble.

3. The method of claim 2, wherein the precipitate is purified by dialysis of an aqueous solution thereof, followed by adsorption chromatography and gel filtration.

4. The method of claim 2, wherein the alkanol is methanol.

5. The method of claim 4, wherein the methanol concentration is about 40 to 80% by weight.

6. A process according to claim 3, wherein the adsorbent for adsorption chromatography is DEAE-cellulose or CM-cellulose.

7. A method of inducing in a mammalian host a substance having the capacity to inhibit plaque formation of vesicular stomatitis virus on RK13 rabbit kidney cell-line culture comprising administering to a mammal the polysaccharide of claim 1 in an amount sufficient to induce said substance.