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WO1995024907A2 - Compositions pharmaceutiques contenant du sulfate de laminarine - Google Patents

Compositions pharmaceutiques contenant du sulfate de laminarine Download PDF

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Publication number
WO1995024907A2
WO1995024907A2 PCT/GB1995/000515 GB9500515W WO9524907A2 WO 1995024907 A2 WO1995024907 A2 WO 1995024907A2 GB 9500515 W GB9500515 W GB 9500515W WO 9524907 A2 WO9524907 A2 WO 9524907A2
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Prior art keywords
heparin
sulfate
laminarin
bfgf
laminarin sulfate
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PCT/GB1995/000515
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English (en)
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WO1995024907A3 (fr
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Israel Vlodavsky
Hua-Quan Miao
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Hadasit Medical Research Services & Development Company Limited
Whalley, Kevin
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Application filed by Hadasit Medical Research Services & Development Company Limited, Whalley, Kevin filed Critical Hadasit Medical Research Services & Development Company Limited
Priority to AU18567/95A priority Critical patent/AU1856795A/en
Publication of WO1995024907A2 publication Critical patent/WO1995024907A2/fr
Publication of WO1995024907A3 publication Critical patent/WO1995024907A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters

Definitions

  • the present invention relates to pharmaceutical compositions containing sodium laminarin sulfate, which have been found to mimic certain heparin activities and which exhibit about 30% of the anti-coagulant activity of heparin, and which therefore can be used instead of heparin for preventing restenosis, accelerating wound healing, and inhibiting tumor cell metastasis.
  • Heparin is a multifunctional, linear, highly sulfated polysaccharide consisting of alternating uronic acid (either L-iduronic or D-glucuronic) and D-glucosamine residues. It exhibits a high degree of heterogeniety due to variation in the size of the polysaccharide chains and the degree and distribution of sulfate groups [see, e.g., U. Lindahl, "Biosynthesis of Heparin and Related Polysaccharides," in: Heparin: Chemical and Biological Properties. Clinical Applications. D.A. Lane and ⁇ . Lindahl, Eds., Edward Arnold, London, pp. 159-189 (1989)].
  • heparin Although heparin is best known for its anticoagulant and antithrombotic properties [see, e.g., J. Hirsh, "Heparin,” New En ⁇ . J. Med.. Vol. 324, pp. 1565-1574 (1991)], it affects a variety of physiological processes such as vascular endothelial and smooth muscle cell proliferation [see, e.g., J.J. Castellot, et al., "Structural Determinants of the Capacity of Heparin to Inhibit the Proliferation of Vascular Smooth Muscle Cells," and "Evidence for a Pentasaccharide Sequence that Contains a 3-O-Sulfate Group," J. Cell Biol.. Vol. 102, pp.
  • laminarin is a polysaccharide found in brown seaweed and occurring principally in the Laminaria spp. It is a linear polymer composed of ⁇ -(l 3)-linked glucose residues. It may contain small amounts of ⁇ -(l 6) linkages as interresidue linkages or as branch points and 2-3% D-mannitol as end groups [Peat, et al., J. Che . Soc. 724, 729 (1958); N. Maeda, Carbohvd. Res.. Vol. 7, p. 97 (1968); Black, et al. in Industrial Gums. R.L.
  • Laminarin can be sulfated to varying degrees [R. I. Whistler, Methods in Carbohydrate Chemistry. Vol. 6, pp. 426-429 (1972)]. Highly sulfated products have anticoagulant properties comparable to heparin, while laminarins with few sulfate groups are antilipemic only [Besterman, Evans, Brit. Med. J.. Vol. I, pp. 310-312 (1957)].
  • Sodium laminarin sulfate exhibits 30-50% of the anticoagulant activity of heparin, and is effective therapeutically in the prevention and treatment of ischemic cerebrovascular diseases [see, e.g., Z.Y. Han, et al. , "Clinical and Laboratory Observations on Polysaccharide Sulfate (PSS) in 282 Cases of Ischemic Cerebrovascular Disease," Clin. Med. J. Engl. , Vol. 104, pp. 562-566 (1991) ] .
  • PSS Polysaccharide Sulfate
  • Laminarin from laminaria digitata is commercially available (Sigma L-9634) . As demonstrated in Fig. 9A, this preparation was not capable of stimulating bFGF-mediated DNA synthesis in F-32 lymphoid cells. It also failed to inhibit melanoma heparanase activity (Fig. 9B) and lung colonization (not shown) . Thus, it would have been assumed by persons skilled in the art that laminarin would not be useful as a mimic of heparin activities. According to the present invention, however, it was discovered that a polysulfated laminarin, having a molar ratio of sulfate group to monosaccharide units (i.e. glucose residues) of at least 1:1 was effective for preventing restenosis, accelerating wound healing and inhibiting tumor cell metastasis.
  • a polysulfated laminarin having a molar ratio of sulfate group to monosaccharide units (i.e. glucose residues) of at
  • laminarin sulfate as used hereinafter is intended to designate such a polysufated laminarin.
  • the molar ratio of sulfate groups to monosaccharide units in laminarin sulfate has been determined to be 1.03:1, whereas laminarin (Sigma) had a ratio of 0.044:1.
  • Sulfate Teho, T.T. & Hartiula, . (1971) Anal. Biochem. 41:471-476
  • glucose hexose
  • the laminarin sulfate (4.2 mg/ml, nominal) of the present invention had a content of glucose of 2.08 mg/ml (i.e. 11.6 mM) and sulfate content of 1.15 mg/ml (i.e. 12.0 mM) .
  • Laminarin from Sigma (10 mg/ml) had a content of glucose of 8.25 mg/ml (i.e. 45.8 mM) and sulfate 0.194 mg/ml (i.e. 2.0 mM) .
  • the polysulfated laminarin used in the present invention is a hydrophylic polymer (Mr « 10,000) consisting of ⁇ -l,3-glucan isolated from the cell walls of seaweeds, which has been subjected to chemical O-sulfation at positions 2 and 6 (2.34 sulfates per monomer), as described by R.L. Whistler, Methods in Carbohydrate Chemistry. Vol. 6, pp. 426-429 (1972); S. Alban, et al., "Synthesis of Laminarin Sulfates with Anticoagulant Activity," Drug Res.. Vol. 42, pp. 1005-1008 (1992) .
  • a pharmaceutical composition comprising laminarin sulfate, for mimicking heparin activity and for therapeutic use instead of heparin in preventing restenosis by the inhibition of vascular smooth muscle cell proliferation, in accelerating wound healing by activating the release of active growth factors stored in the extra- cellular matrix, and for inhibiting tumor cell metastasis by inhibition of heparanase activity; said laminarin sulfate being present in a pharmaceutically effective amount and in combination with a pharmaceutically acceptable carrier, and wherein the molar ratio of sulfate groups to monosaccharide units is said laminarin sulfate is at least 1:1.
  • the invention further encompasses a pharmaceutical composition as hereinabove described, wherein the carrier is a liquid and the composition is a solution.
  • the amount of the laminarin sulfate incorporated in the pharmaceutical composition may vary widely. Factors considered when determining the precise amount are well-known to those skilled in the art. Examples of such factors include, but are not limited to, the subject being treated, the specific pharmaceutical carrier, route of administration being employed, and the frequency with which the composition is to be administered.
  • the compounds of the present invention are administered in a pharmaceutical composition which comprises the compound and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier encompasses any of the standard pharmaceutically accepted carriers, such as a phosphate- buffered saline solution, water, emulsions such as an oil/ water emulsion or a triglyceride emulsion and various types of wetting agents.
  • An example of an acceptable triglyceride emulsion useful in the intravenous and intraperitoneal administration of the compounds is the triglyceride emulsion commercially known as Intralipid.
  • the administration of the pharmaceutical composition may be effected by any of the well-known methods, including, but not limited to, intravenous, intraperitoneal, intramuscular, or subcutaneous administration.
  • Fig. 1 is a characteristic curve showing the effect of the composition of the present invention and of heparin on the binding bFGF to ECM
  • Fig. 2 is a characteristic curve showing the release of ECM- bound bFGF using the composition of the present invention and heparin
  • Figs. 3A,B and 3C,D respectively are characteristic curves and bar graphs, showing the restoration of bFGF receptor binding in heparan-sulfate deficient CHO cells
  • Figs. 4A and 4B, respectively are characteristic curves and bar graphs, showing that laminarin sulfate enables lymphoid cells to synthesize DNA (incorporate H-thymidine) in response to bFGF
  • Fig. 1 is a characteristic curve showing the effect of the composition of the present invention and of heparin on the binding bFGF to ECM
  • Fig. 2 is a characteristic curve showing the release of ECM- bound bFGF using the composition of the present invention and heparin
  • FIG. 5 is a characteristic curve, showing the inhibition of smooth muscle cell proliferation using the composition of the present invention and heparin;
  • Figs. 6A and 6B show characteristic curves for the inhibition of bFGF binding to SMC by laminarin sulfate and heparin;
  • Figs. 7A and 7B are characteristic curves showing inhibition of heparanase activity by laminarin sulfate and heparin;
  • Fig. 8 is a photograph of the effect of laminarin sulfate on lung colonization of melanoma cells;
  • Fig. 8A shows the lungs of an untreated mouse;
  • Fig. 8B shows the lungs of a laminarin sulfate-treated mouse
  • Figs. 9A and 9B, respectively, are characteristic curves showing that commercially available laminarin fails to:
  • FIG. 10 is a schematic representation of the disaccharide repeating unit of laminarin sulfate.
  • Recombinant human bFGF was kindly provided by Takeda Chemical Industries (Osaka, Japan) .
  • Sepharose 6B was obtained from Pharmacia (Uppsala, Sweden) .
  • Sodium heparin from porcine intestinal mucosa was obtained from Hepar Industries (Franklin, Ohio, U.S.A.).
  • Bacterial (flavobacteriu heparinum) heparinase I (EC 4.2.2.7) was kindly provided by Dr. J. Zimmer ann (IBEX Technologies, Montreal, Canada).
  • Heparan sulfate degrading endoglycosidase was purified from human placenta. Enzyme purification involved ammonium sulfate precipitation and sequential chromatographies over carboxymethyl Sepharose, heparin Sepharose and ConA Sepharose.
  • Dulbecco's modified Eagle's medium (DMEM) , Nutrient mixture F-12, RPMI-1640 medium, fetal calf serum (FCS) , newborn calf serum (NBCS) , penicillin, streptomycin, L-glutamine and saline containing 0.05% trypsin, 0.01M sodium phosphate (pH 7.4) and 0.02% EDTA (STV) were obtained from Biological Industries (Beit-Haemek, Israel) . Tissue culture dishes were obtained from Falcon Labware Division, Becton Dickinson (Oxnard, California, U.S.A.). Four well tissue culture plates were obtained from Nunc (Roskilde, Denmark). Na [35S]0.
  • SMC were isolated from the bovine aortic media as previously described [see, e.g., J.J. Castellot, et al. , "Structural Determinants of the Capacity of Heparin to Inhibit the Proliferation of Vascular Smooth Muscle Cells," and “Evidence for a Pentasaccharide Sequence that Contains a 3-O-Sulfate Group,” J. Cell Biol.. Vol. 102, pp. 1979-1984 (1986); and A. Schmidt, et al., "The Antiproliferative Activity of Arterial Heparan Sulfate Resides in Domains Enriched with 2-0-Sulfated Uronic Acid Residues," J. Biol. Che .
  • Glycosaminoglycan deficient Chinese hamster ovary (CHO) cells (pgs A-745) were kindly provided by Dr. J.D. Esko (Department of Biochemistry, University of Alabama at Birmingham, Birmingham, Alabama) .
  • Mutant pgs A-745 cells are deficient in xylosyltransferase, which catalyzes the first sugar transfer step in GAG biosynthesis.
  • the total sulfated GAGs produced by these cells is ⁇ 5% of the amount made by the wild type CHO cells [J.D. Esko, "Genetic Analysis of Proteoglycan Structure, Function and Metabolism," Curr. Opin. Cell Biol., Vol. 3, pp. 805-816 (1991)].
  • PgsA-745 cells were transfected with the cDNA for the murine 2 immunoglobulin-like ectodo ain form of bFGF receptor-l(flg) , yielding pgs A-745 fig cells [A. Yayon, et al., "Cell Surface, Heparin-Like Molecules Are Required for Binding of Basic Fibroblast Growth Factor to Its High Affinity Receptor," Cell. Vol. 64, pp. 841-848 (1991); and D.
  • F32 cells were obtained following transfection of BaF3 cells with Mo/mFRl/SV expression vector and selection in medium containing bFGF plus heparin, as described in D.M. Ornitz, et al., "Heparin Is Required for Cell-Free Binding of bFGF to a Soluble Receptor and for Mitogenesis in Whole Cells," Mol. Cell. Biol.. Vol. 12, pp. 240-247 (1992). Preparation of Dishes Coated with ECM:
  • Bovine corneal endothelial cells were dissociated from stock cultures (second to fifth passage) with STV and plated into 4-well plates at an initial density of 2x105 cells/ml. Cells were maintained as described above, except that 5% dextran T-40 was included in the growth medium and the cells were maintained without addition of bFGF for 12 days.
  • the subendothelial ECM was exposed by dissolving (5 min, room temperature) the cell layer with PBS containing 0.5% Triton X-100 and 20 mM NH.OH, followed by four washes in PBS. The ECM remained intact, free of cellular debris and firmly attached to the entire area of the tissue culture dish [see D. Gospodarowicz, et al.
  • corneal endothelial cells were plated into 4-well plates and cultured as described above.
  • Na_[35S]0 540-590 mCi/mmol
  • was added 40 ⁇ Ci/ml 1 day and 5 days after seeding, and the cultures were incubated with the label without medium change [M. Bar-Ner, et al. , "Inhibition of Heparanase Mediated Degradation of Extracellular Matrix Heparan Sulfate by Modified and Non-Anticoagulant Heparin Species," Blood. Vol. 70, pp. 551-557 (1987)].
  • 10 to 12 days after seeding the cell monolayer was dissolved and the ECM exposed, as described above.
  • ECM was incubated (24 h, 37°C, pH 6.2) with placental heparanase and sulfate labeled material released into the incubation medium was analyzed by gel filtration on a Sepharose 6B column. Intact HS proteoglycans were eluted next to the void volume (Kav ⁇ 0.2) and HS degradation fragments eluted with 0.5 ⁇ Kav ⁇ 0.8.
  • Recombinant bFGF was iodinated using chloramine T, as described in M. Benezra, et al. , "Thrombin-Induced Release of Active Basic Fibroblast Growth Factor - Heparan Sulfate Complexes from Subendothelial Extracellular Matrix," Blood, Vol. 81, pp. 3324-3332 (1993) .
  • the specific activity was 1.2-1.7x105 cpm/ng bFGF and the labeled preparation was kept for up to 3 weeks at -70°C.
  • Confluent SMC or CHO-pgsA-745 fig cells cultured in 24-well plates, were washed twice with binding medium (DMEM.4.5 g glucose/liter, 0.1% bovine serum albumin, 25 mM HEPES, pH 7.4) and incubated with 1251-bFGF (5 ng/ml, 2 h, 4°C) in the absence and presence of increasing concentration of laminarin sulfate or heparin. 1251-bFGF binding to low and high affinity receptor sites on the cell surface was determined, as described by A.
  • binding medium DMEM.4.5 g glucose/liter, 0.1% bovine serum albumin, 25 mM HEPES, pH 7.4
  • 1251-bFGF 5 ng/ml, 2 h, 4°C
  • Cells or ECM were incubated with 1251-bFGF, as described above. Unbound bFGF was washed away and the cells or ECM incubated with heparin or laminarin sulfate at 4°C for 2 h (cells) , or at room temperature for 3 h (ECM) . The incubation media were collected and counted in a gamma-counter to determine the amount of released iodinated material. The remaining cells and ECM were incubated (3 h, 37°C) with IN NaOH, and the solubilized radioactivity counted in a gamma-counter. The percentage of released 1251-bFGF was calculated from the total cell- or ECM- associated radioactivity [R.
  • F32 cells were washed twice with RPMI 1640 medium.
  • Cells (2x104 per well) were plated in 96-well microtiter plates in the absence and presence of increasing concentrations of laminarin sulfate and 5 ng/ml bFGF in a total volume of 200 ml. 48 hours later, 1 ⁇ Ci of 3H-thymidine was added per well, the cells were incubated for another 6 h and then collected with a PHD Cell Harvester [see D.M. Ornitz, et al., ibid.]. Incorporated thymidine was determined by liquid scintillation counting.
  • % Inhibition (1- net growth in presence of laminarin sulfate/net growth in control x 100) .
  • the net growth was determined by subtracting the initial cell number from the final cell number.
  • C57BL mice received a single intraperitoneal injection of low Mr heparin (Fragmin) or laminarin sulfate (400 ⁇ g/0.2 ml/mouse) , 20 min prior to an intravenous innoculation of B16-BL6 melanoma cells (1x105 cells/mouse) .
  • Mice were sacrificed 15 days later, the lungs fixed in Bouen's solution and scored for the number of metastatic nodules [C.R. Parish, et al. , ibid. (1987); M. Nakajima, et al., "Heparanase and Tumor Metastasis," J. Cell. Biochem.. Vol. 36, pp. 157-167 (1988)].
  • ECM coated 4-well plates were incubated (2 h, 24°C) with 1251-bFGF (5 ng/ml) in the absence and presence of increasing concentrations of laminarin sulfate or heparin. Unbound 1251-bFGF was washed away and the remaining ECM-bound bFGF solubilized and counted in gamma-counter. 50% inhibition of 1251-bFGF binding to ECM was obtained in the presence of 0.1 ⁇ g/ml laminarin sulfate, similar to the inhibition obtained in the presence of 0.1 ⁇ g/ml heparin (Fig. 1).
  • ECM was incubated (2 h, 24°C) with 1251-bFGF, washed free of unbound bFGF and exposed (3 h, 24°C) to increasing concentrations of laminarin sulfate or heparin.
  • half maximal release of the ECM-bound bFGF was achieved in the presence of as little as 0.1 ⁇ g/ml laminarin sulfate, similar to the amount of bFGF released under the same conditions by 0.1 ⁇ g/ml heparin.
  • laminarin sulfate efficiently released 1251-bFGF that was first bound to low affinity HS binding sites on the surface of vascular SMC (not shown) .
  • addition of laminarin sulfate resulted in a marked stimulation of 3H-thymidine incorporation in F32 cells exposed to bFGF.
  • a maximal effect was obtained in the presence of 0.2 ⁇ g/ml laminarin sulfate (Fig. 4A) , similar to the effect of heparin observed under the same conditions (Fig. 4B) .
  • heparin oligosaccharides prepared by limited digestion with nitrous acid retain antiproliferative activity and that both the N/O-sulfate ratio as well as size are important in determining antiproliferative activity [J.J. Castellot, ibid. ] .
  • Sparsely seeded vascular SMC were exposed to 10% FCS in the absence and presence of increasing concentrations of laminarin sulfate or heparin.
  • a similar dose dependent inhibition of cell proliferation was obtained with both polysaccharides (Fig. 5) .
  • 50% inhibition of cell proliferation induced by 10% FCS was obtained in the presence of 40 ⁇ g/ml laminarin sulfate and 100 ⁇ g/ml heparin (Fig. 5).
  • a necessary component of the metastatic process is the ability of tumor cells to bind to and degrade basement membranes and ECM [L.A. Liotta, et al., "Tumor Invasion and Extracellular Matrix,” Lab. Invest.. Vol. 49, pp. 636-649 (1983)].
  • ECM extracellular Matrix
  • Heparanase activity correlates with metastatic potentials of mouse lymphoma, melanoma and fibrosarcoma cells [M. Nakajima, et al., ibid. ; I. Vlodavsky, et al., ibid. ; M. Nakajima, et al., "Heparan Sulfate Degradation: Relation to Tumor Invasion and Metastatic Properties of Mouse B16 Melanoma Sublines," Science. Vol. 220, pp. 611- 612 (1983); T.
  • Elevated levels of the enzyme were detected in sera from metastatic tumor-bearing animals and melanoma patients and in tumor biopsies of cancer patients [M. Nakajima, et al., ibid. (1988); I. Vlodavsky, et al., "Involvement of Heparanase in Tumor Metastasis and Angiogenesis," Is. J. Med.. Vol. 24, pp. 464-470 (1988); and T. Peretz, et al., "Maintenance on Extracellular Matrix and Expression of Heparanase Activity by Human Ovarian Carcinoma Cells from Biopsy Specimens," Int. J. Cancer. Vol. 45, pp. 1054-1060 (1990) ] .
  • Degradation of HS in the ECM is studied by allowing cells or purified enzyme preparations to interact with a metabolically sulfate labeled ECM produced by cultured bovine corneal endothelial cells. Sulfate labeled degradation products released into the incubation medium are analyzed by gel filtration on Sepharose 6B column [M. Bar-Ner, et al., ibid. (1987); M. Nakajima, et al., ibid. (1988); I. Vlodavsky, et al., ibid. (1983)].
  • HSPG While intact HSPG is eluted next to the void volume of the column, labeled degradation fragments of HS side chains are eluted more toward the Vt of the column (0.5 ⁇ Kav ⁇ 0.8, Mr - 4-7x103) Heparanase mediated degradation of HS is inhibited by heparin, regardless of whether intact cells or purified enzymes are incubated with the ECM. Heparanase purified from human placenta was incubated (24h, 37°C, pH 6.2) with sulfate labeled ECM in the absence and presence of increasing concentrations of laminarin sulfate or heparin.
  • Fig. 8B demonstrates that a single intraperitoneal injection of laminarin sulfate (400 ⁇ g/mouse) prior to an intravenous inoculation of B16-BL6 melanoma cells to C57BL mice, decreased the incidence of melanoma lung metastases to less than 5% of the number of metastatic nodules found in control untreated mice (Fig. 8A) .
  • a similar inhibition of melanoma lung metastasis was observed in C57 BL mice innoculated with B16-BL6 melanoma cells and low Mr Heparin (Fragmin, 400 ⁇ g/ mouse) .
  • glycosaminoglycans GAG
  • HS heparin and heparan sulfate
  • Such interactions range from highly specific, lock-and-key type binding, as described for the antithrombin binding region in heparin [U. Lindahl, et al., ibid. (1989)], to relatively nonspecific, co-operative electrostatic association [A.D. Cardin and H.J.R. Weintraub, ibid. (1989) ; H. Margalit, et al. , ibid. (1993) ] .
  • laminarin sulfate was, on a molar basis, more effective than heparin. Low concentrations of either laminarin sulfate or heparin inhibited the binding of bFGF to HS binding sites on ECM and cell surfaces. These polysaccharides were, on a weight basis, equally effective in displacing ECM- or cell-bound bFGF. Moreover, laminarin sulfate induced restoration of bFGF receptor binding and mitogenic activity in HS deficient CHO cells [A. Yayon, et al. , ibid. (1991); J.D.
  • bFGF binds preferentially to HS regions enriched in Ido A(2-OSO ) ⁇ l-4 GlcNSO- and that a minimum size of 8-10 sugar units is required for a heparin-derived olgosaccharide to support the mitogenic activity of bFGF [D.M. Ornitz, et al., ibid. (1992); D. Aviezer, et al., ibid. (1994); J.E. Turnbull, et al., ibid. (1992); D.J. Tyrrell, et al., ibid. (1993); and M. Maccarana, et al., ibid. (1993)].
  • N- sulfation of glucosamine and 2-O-sulfation of iduronic acid are required for bFGF recognition, whereas 6-O-sulfation does not influence the interaction.
  • laminarin sulfate an unrelated polysaccharide (laminarin sulfate) was found to be as effective as heparin in its ability to restore bFGF receptor binding and mitogenic activity, as well as to inhibit the proliferation of vascular SMC. It is conceivable that oversulfation such as that found in laminarin sulfate may provide the necessary density and/or distance between sulfate groups and thus compensate the need for specific structural determinants that mimic the type of interactions sufficient to elicit the biological responses induced by heparin and HS. Analysis of the exact mode of interaction between bFGF and laminarin sulfate must await a better characterization of the structure and sulfation pattern of laminarin sulfate.
  • bFGF receptor binding and activation is also induced by several other polysaccharides (i.e., pentosan sulfate, dextran sulfate), but not by other sulfated compounds (i.e., sucrose octasulfate, polyanetholesulfonic acid) .
  • sulfated polysaccharides i.e., heparin, fucoidan, carrageenan lambda, pentosan polysulfate
  • others are either not effective (i.e., chondroitin-4- sulfate, chondroitin-6-sulfate, carrageenan kappa, hyaluronic acid) or partially effective (i.e., dextran sulfate) [C.R. Parish, et al., ibid. (1987)].
  • the potent anti-metastatic activity of laminarin sulfate is therefore not an obvious property characteristic of a wide variety of sulfated polysaccharides. Moreover, while the above-mentioned anti-metastatic polysaccharides were less active than heparin [D.R. Coombe, et al., "Analysis of the Inhibition of Tumor Metastasis by Sulfated Polysaccharides," Int. J. Cancer. Vol. 39, pp. 82-88 (1987)], laminarin sulfate was even more potent than heparin in inhibiting heparanase activity and lung colonization, while exerting a much lower anticoagulant activity.
  • the present invention provides new clinical applications of laminarin sulfate, apart from its current use in the prevention and treatment of cerebrovascular thrombosis.
  • Laminarin sulfate can be used in a pharmaceutical composition to accelerate wound healing and neovascularization by virtue of its ability to release active heparin binding growth factors and to promote bFGF receptor binding and mitogenic activity. Its ability to suppress SMC proliferation can be utilized in the inhibition of restenosis and accelerated atherosclerosis following transluminal balloon de-endothelialization.
  • Laminarin sulfate may also reduce the incidence of cell invasion in tumor metastasis and autoimmunity by virtue of its efficient inhibition of heparanase activity and the related extravasation of normal and malignant blood borne cells.

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Abstract

L'invention concerne une composition pharmaceutique contenant du sulfate de laminarine de sodium servant à imiter l'activité de l'héparine, et utilisé à des fins thérapeutiques à la place de l'héparine dans la prévention de la resténose par inhibition de la prolifération des cellules des muscles lisses vasculaires, dans l'accélération de la cicatrisation par activation de la libération de facteurs de croissance actifs emmagasinés dans la matrice extracellulaire, ainsi que dans l'inhibition de la métastase de cellules tumorales par inhibition de l'activité d'héparanase. Cette composition pharmaceutique contient une quantité pharmaceutiquement efficace de sulfate de laminarine de sodium combinée à un véhicule pharmaceutiquement acceptable, le rapport molaire entre les groupes de sulfate et les unités de monosaccharide dans ledit sulfate de laminarine étant d'au moins 1:1.
PCT/GB1995/000515 1994-03-13 1995-03-10 Compositions pharmaceutiques contenant du sulfate de laminarine WO1995024907A2 (fr)

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FR2900577A1 (fr) * 2006-05-04 2007-11-09 Goemar Lab Sa Nouveaux medicaments pour les traitements contre le virus de l'herpes

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WO1998003573A1 (fr) * 1996-07-17 1998-01-29 Biomolecular Research Institute Ltd. Composes inhibiteurs angiogeniques
US6426067B1 (en) 1996-07-17 2002-07-30 Biomolecular Research Institute, Ltd. Angiogenic inhibitory compounds
US6440445B1 (en) 1996-09-30 2002-08-27 Brigham & Women's Hospital Methods and compounds for treatment of abnormal uterine bleeding
WO2002036132A1 (fr) * 2000-11-03 2002-05-10 Laboratoires Goemar S.A. Medicament anti-inflammatoire et cicatrisant a base de sulfate de laminarine
US7008931B2 (en) 2000-11-03 2006-03-07 Laboratoires Goemar S.A. Anti-inflammatory and healing medicine based on laminarin sulphate
WO2002096945A3 (fr) * 2001-05-24 2003-02-20 Isis Innovation Recepteur des macrophages
WO2003045414A3 (fr) * 2001-11-30 2003-10-16 Goemar Lab Sa Traitements therapeutiques
FR2900577A1 (fr) * 2006-05-04 2007-11-09 Goemar Lab Sa Nouveaux medicaments pour les traitements contre le virus de l'herpes
WO2007128914A1 (fr) * 2006-05-04 2007-11-15 Laboratoire De La Mer Nouveaux medicaments pour les traitements contre le virus de l'herpès
US8288363B2 (en) 2006-05-04 2012-10-16 Ase & Bio Medicaments for anti-herpes virus treatments

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