Formulation and biological activity of antineoplastic proteoglycans derived from Mycobacterium vaccae in chitosan nanoparticles.
Although heat-killed suspensions of Mycobacterium vaccae have been tested clinically against tuberculosis and cancer, from a pharmaceutical perspective it would be advantageous to utilize isolated active components rather than the heat-degraded bacterial materials. In our laboratory we have isolated from M. vaccae a number of high-molecular-weight proteoglycans with considerable immunological and antineoplastic activity. The structure of one of these, PS4A, obtained by extraction with boiling water, seems to consist of a basic unit with a 20-kDa protein core to which are attached glucans and O-methylated 4-kDa polysaccharides. The molecular weight is (approx.) 50 kDa, but because of self-association, that of the recovered high-molecular-weight fraction is greater than 150 kDa. A similar, but even larger, molecule (PS4alpha, MW approximately 20 MDa) is obtained by cold extraction with 8 M urea. Both are active in-vivo against an S-180 murine sarcoma model but have no activity in-vitro, suggesting an antitumour effect involving activated macrophages. For this reason gelatin nanoparticles are unsuitable as a vehicle but chitosan seemed to be a promising alternative. In this report we describe the production of stable 600-700-nm diameter nanoparticles of chitosan without organic solvents. Adsorption and release of bovine serum albumin seemed to be affected by the charge of the two reactants and at high doses not all adsorbate was released. PS4A, because of structural and compositional differences, had to be loaded on to the chitosan by freeze drying a suspension of the nanoparticles in a solution of the drug. After a rapid (burst) release phase, the rate of release into water was steady for the next 4 h, but not all the drug was released. In-vivo it was evident that PS4A and PS4alpha were equally active in solution or when formulated in the chitosan nanoparticles. These results show that chitosan nanoparticles, readily prepared without the use of organic solvents, are a suitable vehicle for the delivery of these immunostimulants from M. vaccae; the formulations might find application as antitumour agents.
Tian
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Institute for Tuberculosis Research, College of Pharmacy, University of Illinois at Chicago, 60607, USA.
Groves
M J
MJ
D000818:Animals; D000970:Antineoplastic Agents / Q000302:isolation & purification* / Q000494:pharmacology; D002417:Cattle; D002626:Chemistry, Pharmaceutical; D002686:Chitin / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000648:ultrastructure; D048271:Chitosan; D051379:Mice; D017348:Microscopy, Electron, Scanning Transmission; D009161:Mycobacterium / Q000737:chemistry*; D010316:Particle Size; D011135:Polysaccharides, Bacterial / Q000302:isolation & purification / Q000494:pharmacology; D011509:Proteoglycans / Q000302:isolation & purification* / Q000494:pharmacology; D012510:Sarcoma 180 / Q000188:drug therapy / Q000473:pathology; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000493:pharmacokinetics; D013997:Time Factors
D000970:Antineoplastic Agents; D011135:Polysaccharides, Bacterial; D011509:Proteoglycans; D002686:Chitin; D012710:Serum Albumin, Bovine; D048271:Chitosan