Title of Invention	MINIMAL SULFATED POLYSACCHARIDES
Abstract	ABSTRACT 349/CHENP/2005 The present invention relates to a minimal sulfated polysaccharide having a percentage of sulfur above 6% and below 13% with respect to the simple sugar residue and wherein the molecular weight is above 5 000 g/mol.

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1. FIELD OF THE INVENTION Tins invention reiaics to methods for treating or preventing microbial infections in mammals using sulfated polysaccharides. Mors particularly, this invention relates to methods of introducing a therapeutically effective amount of a charged and flexible sulfated polysaccharide having a certain percent sulfation range into the blood stream, lymphatic system and/or extracellular spaces of a human patient for the treatment, prevention or management of microbial infections, In particular, wherein the range is effective to enable maximal interaction of the sulfate groups with the microbe which causes the infection, and wherein the sulfated polysaccharide is not substantially endocytosed or degraded by cell receptor binding in the mammal, and thereby retains antimicrobial activity in vivo. 2. BACKGROUND OF THE INVENTION Charged polysaccharides, particularly sulfated polysaccharides, ha.ve demonstrated potent antimicrobial activities in vitro. (Baha et al Antiviral Res 9L33S-343, 1988; Ito et ai., AnttviralRes. 7{36):l-367, 1987). For example, sulfated polysaccharides such as dextran sutfate, heparin, and pentosan polysulfate have been reported to be potent inhibitors of HIV, paramyxoviruses, cytomegaloviruses, influenza viruses, semlikiviruses (Luschcr-Mattlie et al.. Arch Virol 130J17-326, 1993) and herpes simplex viruses in viro (Baba et ai., Andmicrob. Agents Chemotherapy 32:1742-45, 1988: Pancheva, Antiviral Chem Chemotherapy 4;189191, 1993), However, these known compoundi have disappointtngly poor activity in vivo. Dextcan. sulfate and heparin 'were first reported to inhibit HIV eplication in vitro by Ito et ai.. Antiviral Res. 7:3(51-367,1987, Deringer er ai (US 5,153,181) and Ueno and Kuno Lancet 2:796-97, 1987. Later, several other sulfated polysaccharides were shown to inhibit HIV replication at concentrations believed to be below their respective cytotaxcicty thresholds, e.g., pentosan sulfate (Baba etaL, Antiviral Res 9:335-343, 19SS; Biesert et a!., Aids 2(6):449-57. 1988), fudodan (Baba ei aj.. Antiviral Res 9:335-343, 1988), lambda-, kappa- and iota-carrageenan (Baba et al., Antiviral Res 9: 335-343, 1988), lentinan sulfate fYoahida et aL.Biockem- Pharmacol 37(15):2gg7-91, 1988), mannan sulfate pto et al, Eur. J. Clin. Microbiol Infect. Dis. S; 19M 93, 1989), dextrin sulfate (llo el al Antiviral Chem. Chemother.. 2:41-44. \99lXsa\£osvemBn(Wei\sr eiaL, J Gen P7ro/71:1957-1963, herpes virus as in EP Application 0 066 379 A2 with limited success. See also, Pancheva SN, Antiviral Chem Chemotherapy 4:189-191, 1993. One of the major reasons that dextran sulfate may pot be active in vivo is that the material is not stable. Some indicationof this has been published previously. Tritium labeled dextran sulfate mw 8,000 appeared to be depolymerised while in the blood circulation of rats over a 6-24 h pcriod (Hartman KR, lohas DG, Mitsuya H. AIDS Res Hum Retroviruses 6: S05-S1!, 1990). lodinated heparin and pentosan polysulphatc are rapidly cleared from the circulation in man and returned in a desulfated fenn (MacGregor IR, Dawes J, Paton L, Pepper DS, Prowse CV, Smith M., Tkramb Haem 51 ;321 -325, 1984). Considerablc effort has been focused on improving the in vivo anti-viral activity of dextran sulfate by increasing its sulfation or modifying the use of conveotional material. In one study, given the reported poor absorption of oral dextran sulfate, dextran sulfate was administered to a maximally tolerated dose by continuous infusi on to subjects with symptomatic HTV infection for up to 14 days. (Flexner et al/.. Antimicrob Agents Chemotherapy 35:25A4-25$0, 1991). Continous intravenous infusion of dextran sulfate was found to be toxic. The authors concluded that as a result of its toxicity and lack of any demonstration of beneficial effect in vivo, dextran sulfate is unlikely to have a beneficial effect in the treatment of HIV. Id. Indeed, the authors cautioned: "further clinical dcvelopment of parental dextran sulfate as therapy forsymptomatic, HIV infection is not warranted and could prove to be hazardous. On the basis of the results of this study, caution is advised in the clinical evaluation of other polysnliatcd polyanions." (Id. at 2549). In a major study of the processing of dextran sulfete by glomerular endothelial cells. Applicant discovered that dextran sulfate binds to a cell surfece receptor that would normally recognize highly sulfated polysaccharides such as heparin-like polysaccharides. On binding the dextran sulfate is endocytosed, desulfated but not depolymerised by lysosome sulfeteses and exocytosed as desulfeted dextran sulfate (Vyas et al. Arch Biochem Biophys. 332(2):20S 12,1996). It was found that the uptake and cndocytosis of dextran sulfete by the eel) was critically depMideot on the sulfur content or degree of sulfate substitution per glucose residue. Above 13% sulfur uptake by glomercular endothelial cells wassignificant where as below 13% sulfur uptake by glomerular endothelial This means that charged polysacchari des with a particular critical sulfater content or critical sulfate substitution charge density along the polysacdiaride chain may be processed differently by cells to which the circulation is exposed. Any organ in the body, particularly the simple sugar residue of greater than 6% and less than l3%, preferably greater than about 7% and less than 13%, more preferably greater than about 9% and less than 13%, most preferably 6%, 7%, 8%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.2%, 12.5%, 12.8% or 12.9%. The sulfated polysscohflrides are preferably suifated dextrans having an a-1,6-glycosidic linkage. The invention further encompasses eulfaled polysaccharides having a molecular wcight between 500 and 1,000,000, preferably above 5,000; more preferably above 25.000; most preferably above 40,000 particularly for oral or parential administration. Ranges of 5.000 to 1.000,000, 25,000 to 500,000 and 40,000 to 300.000 are also encompassed by the invention. However, for topical administration, the sulfated polysaccharide may have a molecular weight higher than 500,000 in a preferred embodiment. In an alternative anbodiment, the composition has only about 10% variability in the molecular weight and preferably about 5% variation. hi a preferred embodiment of the invention, the sulfated polyEaccharide is not cellulose sulfate, dextrin sulfate or cyclodextrin, but instead is an QF-l,6-sulfated polysaccharide such as a sulfated dextran having a controlled range of sulfttion, and, optionally, a specific molecular weight range. In an alternative embodiment, the sulfated polysaccharide is homogenous with respect to molecular weight, percent of sulfation or both. In one aspect of the invention there is provided a method for introducing a therapeutically effective amount of a sulfated polysaccharide or salt thereof into the blood stream, lymphatic system and/or extracellular spaces tissue of amancunal comprising administering to the mammal at least one sulfated polysaccharide or a pharraaceutically acceptable salt or hydrate thereof having antimicrobial activity in vitro and having a percent of sulfation sufficient for retention of the anti-microbial activity in vivo. Prefably, the range of sulfation of the polysaccharide is effective to enable maximal interaction of constituent sulfate groups with the miraobe which causes the infection, and wherein the sulfated polysaccharide is not substantially endocytosed or degraded by cell receptor binding In the mammal, and thereby remains antimicrobial activity in vivo. In another aspect of the invention there is provided a method for treating or preventing a micmbial infection comprising administering to a patient a ther^eutically effective amount of sulfated dextran having a percent of sulfur greater than 6% and below 13%. In a preferred embodiment, sulfated dcxtran has aperient sulfation of above 6% or about or above; 6.5%, 7%, 7.5%, 8%, S.5%, 9%. 9.5%, 10%, 10.5%, 11%, 11.5%, 12%. 12a%, 12.5%, 12.8% or less than 13%. In a preferred embodiment the method is for treating or preventing a viral infection, including but not limited to DNA viruses and RNA viruses, particarly enveloped viruses whether DNA or RNA viruses. In a seperatee and prefected method the virus to be treated include hut are not limited to double-stranded DNA viruses, DNA reverse transcripition viruses, RNA reverse transcripting viruses, double-stranded RNA viruses, negative-sense single stranded RNA viruses, and positive-sense single-stranded RNA viruses. In yet another aspect of the invention, there is provided a method for synthesizing a polysaccharide, or decreasing or increasing the degree of sulfation such that the sulked polysaccharides arc suitable for administration in vivo and are efficacious in vivo agantst viral infection. The method comprises providing the sulfated polysaccharides with a percent of sulfation suffcient to eliminate or reduce binding and intemalizstioa of the sulfated polysaccharides by high charge density polyanion cell receptors, or otherwise inactivate these coompound in vivo but sufficient to provide antimicrobial activity; and administering the sulfated polysaccharide to a mammal. In other words, the invention encompasses modifying the sulfation of a naturally occurring or commercially available sulfated polysaccharide to a range of sulfation effective to enable maximal interaction with the microbe and wherein the sulfated polysaccharide is not substantially cndocytosed or degraded by cell receptor binding. Separate aspects of the invention encompass pharmaceutical compositions suitable for parenteral administration to a patient comprising a therapeciticallyor phannaceutically acceptable amount of a sulfated polysaccharide of the invention; pharmaceutical compositions suitable for oral adnunistration to a patient comprising a therapeutically or phamiaccutically acceptable amount of a suifetcd polysaccharide of the invention; and pharmaceutical compositions suitable for topical administration to a patieni comprising a therapeutically or phannaceutically acceptable amount of a sulfated polysaccharide of the invention having a molecular weight grcater than 500,000. It should be noted that the invention also encompasses the use of the sulfated polysaccharides of the invention as disinfectants that can be used to disinfect inanimate objects in hospitals, laboratories, lavatories, auditorium, stadium, convention centers, restaurants, fitness centers, subway terminals, bus terminals, airports, post offices, offices, sewage treatment facilities, sewerS, water treatment facilities, pumping stations, automobiles, airplanes, trains, homes, lockers, and furniture to prevent the spread of viruses or disease. The invention also encompasses disinfectant compositiona such as solutions, sprays, soaps, foam comprising one or more of the sulfated polysaccharides described herein. The microbial infection encompassed by the methods of the invention, particularly the specific viruses to be treated and specific sulfated dextrans to be used, are described in detail below. 3.1 DEFINITIONS As used herein, the terms patient or subject " mean an animal (e.g., cow, horse, sheep, pig. chiciten, turkey. quail, cat, dog, mouse, rat, rabbit, guinea pig, etc.). preferably a mammal such as a non-primate and a primate {e.g., monkey and human), most prcferably a human. In certain embodiments, the patient is an infant, child, adolescent, adult or geriatric patient. In addition, the patient includes immunocompromised patients such as HTV positive patients, cancer patients, and patients undergoing immunotherapy. As used herein, a "therapeutically effective amount" refers to an amount offhe compound of the invention or other active ingredient sufficient to provide a benefit in the treatment or management of the disease, to delay or minimize symptoms associated with the disease, or to cure or ameliorate the infection or disease or causes thereof. In particnlar, a therapeutically effective amount means an amount sufficient to provide a therapeutic benefit in vivo. Further, a therapeutically effective amount means an amount of a compound of the invention alone, or in combination with other therapies, that provides a benefit in the treatment or management of the disease, to delay or minimize symptoms associated with the disease, or to cure or ameliorate the infection or disease or causes thereot Additionally, a thcrapeutically effectivc means an amount of therapeutic agent thai provides a benefit in the treatment or management of the infection or disease without being toxic to the patient. Used in connection with an amount of a compound of the invention, the term encompasses an amount that improves overall therapy, reduces or avoids symptoms or causes of disease, or enhances the therapoutic efficacy of or synergies with another therapeutic agent. As used hcrein, a ""prophylactically effective amount" refers to an amount of a compound of the invention or other active ingredient suffcient to resuh in the prevention of recurrence or spread of the infection or disease. A prophylacticaliy effective amount may refer to an amount sufficient to prevent initial infection or initial disease or the recurrence or spread of the infection or disease at the occurrence of the disease in a patient, including but not limited tothose predisposed to the disease. In particular, a prophylactically effective amount with respect to a compound of the invention means an amount sufficient to result in the prevention of recurrence or spread of the infection or disease m vivo. A piophylactically effective amount may also refer to an amount that provides a benefit in the prevention of the infection or disease without being toxic to the patient. Further, a prophylactically effective amoont with respect to a compound of the invention means an amount alone, or in combination with other agents, that provides a prophylactic benefit in the prevention ofthe infection or disease. Used in connection with an amount of a compound of the invention, the term encompasses an amount that improves overall prophylaxis or enhances the prophylactic efficacy of or synergies with another prophylactic or therapeutic agent. As used herein, "in combination' refers to the use of more than one prophylactic and/or therapeutic agents simultaneonsly or sequentially and in a manner that their respective effects arc additive or synergistic. As used herein, the terms "manage", "managing" and "management" refer to slowing or preventing the progression or worsening of the disease but not curing the disease. As used herein, the terms "prevent", " preventing" and "prevention" refer to the prevention of the onset, recurrence, or spread of the disease in a subject resulting from the administration of an active ingredient before the disease or infection occurs. As used herein, the terms "treat", "treating" and "Ireatment" refer to the eradication or amelioration of the disease or infection itself causes of the disease or symptoms associated with the disease. In certain embodimcnis, such lemas refer te minimizing the spread or worswiing of the disease or infection resulting from the administration of one or more prophylactic or therapeutic agents to a sublject with such a disease or infection. As used herein, the term "phsrmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable aon- toxic acids or bases including inorganic acids and bases and organic acids and bases. Suitable pharmaceutically acceptable base addition salts for the compound of the present invention include, but are not limited to, metallic salts made from aluminum, calcium, Jifeium, magnesium, potassium, sodium and zinc or organic salts made from lysine, N,N'-dibenzyletfaylenediamine, chloroprocaine, choline, diethanolamine, ethylraiediamine, meglumine (N-methylglucamine) and procaine. As used herein and unless otherwise indicated, the term "optically puce" or "stereomerically pure" means a composition that comprises one stereisoner of a respect to each simple sugar residue within the polysaccharide in question, optionally including a counterion, e.g., molecular weight of sulfation in the composition/total weight. In a preferred embodiment, the percent of sulfur ia calculated as the percent of sulfur by molecule weight with respect to the sulfated sugar residue within the polysaccharide in question with sodium aa the counterion. The percent of sulfation can be determined by element analysis ofmaterial which has been dialyzed to remove free sulfur, preferably of moisture/volatage free material dried in vacuo at 60°C to a constant weight Other methods of detemining percent ofsulfation are via moisture content analysis and titration. Sulfation is to be distinguished from "degree of substitutian*' or "eqiuvalents" -which is a measure of the number of sulfate groups per sugar moiety. However, it will be recognized by one of skill in the art that percent sulfation can be convated to a degree of substitution equivalents and vice versa. As used herein, the tenn "co-charged dextran polyamions is dextran substituted to varying degrees with any combination of carboxymethyl groups, sulfate groups and sulfonate groups. As used herein, the tenn "periodate treated anionic polysaccharides" means any anionic polysaccharide that has been treated with periodate to open the sugar ring without depolymerization or to otherwise increase the flexibility of the polysaccharide in order to increase interaction with the microbe. As used herein, the term "antimicrobial" includes and viral; antibacterial, such as, for example, antichlamydial; antiparasitic, such as aotl-Plasmodium or anti-fimgal, 4. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing the amount of desulfation of circulating plasma commercial dextran sulfete as a function of time in plasma (ii""3) in Sprague-Dawley rats. Values up to 24 hours were based cm dextran sulfate existing in plasma after a bolus intravenous injection at zero time. The mean value at 168 hours was obtained from Steady state osmotic pumps implanted subcutanously in Sprague-Dawley rats. Figure 2 is a graph showing the effective antiviral active concentration of polysaccharide materia} versus time after bolus iv injection (172mg/fc£) at time zero. Group 1: commercial dextran sulfate,mW=40,000(ii=l-3); Group 2: commercial dextran sulfate, mw=500,000 Cn=3); Group 3: sul&ted dextran 12.6% (DES 6 40k) (N=4-6); group 4: dextran sulfate eliminates or at least siginificantly reduces the binding and internalization of of sulfation of greater than 60% and below 13% preferbly than about 7% and below In a separate embodiment the invention encompasss a method of treating or preventing a microbial infection in mammals comprising administering to a mammal in need thereof a therapeutically effective amount of a composition comprising a sulfated polysaccharide having a percent of sulfate substitution-glucose residue in the polysaccharide ranging from greater than 6% to lese than 13%, wherein the range of percent sulfation is effective to enable maximal interaction of constituent sulfate groups with the microbe which causes the infecuon, and wheron the sulfated polysaccharide is not substantially cndocytosed or degraded by cell receptor binding in the inammal, and thereby retains antimicrobial activity in vivo. Preferably, the sulfeled polysaccharide is sui&ti^ dextran. The invention also encompasses the treatment, prevention or management of anli-inflammatory diseases or disorders, interstitial cystisis and and-arthritic diseases. The invention also encompasses the use of the sulfated polysaccharides of the inventionanti-albuminuric agents (albuminuria that occurs in kidney disease). The invention further encompasses a method of treating or preventing a microbial infection in a mammal which comprises administering to a mammal in need thereof ao effective amount of a levorotatory sulfeted poIysaccharide having a percent of sulfation from about 6% to about 20%; preferably from about 6% to about 13%; more preferably from about 9% to about 13%. In a further embodiment, the mvention cncompasses a method of treating or preventing a microbial infection in a mammal which comprises administering to a mammal in need thereof an effective amount of a periodate-treated anionic polysaccharide. Preferably, the pcriodatc treated anionic polysaccharide is a periodate treated sulfated dcxtran. In anotficT embodiment of the invention, the invention encompasses a method of treating or pnsvcnting a microbial inibction in a mammal which comprises administering to a mammal in need of such treatment or prevention an effective amount of a co-charged anionic polysaccharide which has a percent of sulfation which enables maximal interaction ■mth the microbe and which is not substantially endocytosed or degraded by cell receptor binding in the mammals thereby retaining antimicrobial in vivop. In a preferred embodinlent, the co-charged anionic polysaccharide is co-charged with carboxymethyl groups, sulfonate groups, sulfete gmups or mixtures theieof; more preferably the co-charged anionic polysaccharide is co-charged with carboxymethyl groi^- In a specific embodimeait, the co-charged anionic polysaccharide is carboxymethyl dextran sulfete or caihoxymethyl cellulose. (fish), Marek's disease virus (flow) Movar herpesvirus, Myxoma virus, Orf virus (contagious pustular dennatitis virus), Pseudocowpox virus (milker's nodule virus). calicivirus, Human astrovimses 1-5, Bovine astrovirases 1-2, Ovine astrovims. Porcine astrovirus Canine astrovims, and Duck astrovims. Specific enveloped single-stranded positive sense RNA viruses which can be treated, prevented or managed by the methods of the present invention include but are nott limited to Barmah Forest virus. Central European encephalitis virus, Chikungimya vims. Dengue viruses 1-4, Eastern equine encephalitis virus, Hepatitis C virus, Human immunodeficiency viruses 1 and 2, Human T-iymphotropic viruses 1 and 2, Igbo Ora virus, Japanese encephalitis virus. Kyasanur forest virus, Mayara virus, Murray Valley encephalitis vims, O'nyong-nyong virus, Omsk hetaoniiagic fever virus, Rocio ■^'inis, Ross RivCT vims, Kubella virus. Ru^ian spring-summer encephalitis virus, SemliJd Forest virus, Sindbis vims (and variants Ockelbo and Babanki vimsea), SL Loius encephalitis virus, Venezuelan equine encephalitis vims. West Nile vims. Western equine encephalitis virus. Yellow fever virus, Avian reticuloendotheliosis virus, Avian sarcoma and leukosis viruses. Border disease virus (sheep). Bovine inununodeficieDcy virus. Bovine leukemia vims. Bovine diarrfiea virus. Caprine arthritis-encephalitis viruB, Classical swine fever vims, Eastern equine sacepbalitis virus. Equine infectious anemia virus. Feline immunodeficjaicy virus, Fehne leukemia virus. Feline sarcoma virus, Getah virus. Hog cholera virus, Japanese encephalitis virus. Lactic dehydrogenase-elevating virus (mice), Maedi/viana virus (sheep). Mouse hepatitis viruses, Mouse mammary tumor virus, Mucosal disease virus (cattle). Murine leukemia viruses (including Abelson, AKR, Friend, Maloney leukemia viruses, Progressive pneumonia virus of sheep, Rous sarcoma virus, Rauscher murine leijkemia virus, Simian Immunodeficiency viruses (including African Green Monkey, Sooty mangabey, Simian-tailed macaque, pig-tailed macaque, Rhesus, Chimpanzee, and Mandrill viruses), Simian Type D retrovirus. Simian T-cell lymphotrophic viruses. Tick-borne encephalits viruses (including European aod far eastern tick-borne encephalitis viruses, Louping ill virus, and Powassao virus), Venezuelan equine ajccphalitJs virus, Wcsselsbron virus, and Western equine encephalitis vims. Woolly monkey sarcoma vims. Specific enveloped single stranded negative sense RNA viruses which can be treated, prevented or managed by the methods of the present inveotion include, but arc not limited to Alagoas virus, Bunyamwera virus, Bwaaba virus, California encephalitis virus, Congo-Crimean hcmonhagit fever vims, Chandipura vitus, Duvaihage virus, Guama vims, Guanarilo vims, Hantaan virus. Influenza vimsra A, B, and C, isfahan virus, Jamestown Canyon virus, Junin virus (Argeatine hemorrhagic fever virus), Lagos bat virus. La Crosse virus, Lassa virus, Lymphocytic choriomeningits virus (LCM virus), Machupo virus, Maraba virus, Marburg virus. Measles virus, Mumps virus, Mokola vims, Muerto Canyon virus, Oriboca virus, Oiopouche virus, Parainfluenza viruses 1 (Sendai virus), 2, 3. 4a, and 4b, Piohinde virus, "Piry virus, Puuto toro virus, Puumaia virus. Rabies virus, Respiratory syncytial virus. Rift Valley feva- virus, Sandfly fever-Kaples virus. Sandfly fever-Sicilian virus, Seoul vims. Sin Nombre virus, Tacaribc virus, Tahyna virus, Tamiami virus. Vesicular stomatitis vinises (including New Je«ey and Indiana strains), Akabaoe virus, Aino virus, Avian paramyxovirus 2 (Yucaipa virusX 3, 4, 5 (Kunitachi virus), 6, 7, 8, and 9, Bcfvine ephemeral fever virus. Bovine respiratory syncytial virus. Canine distemper virus, Dobbin and Porpoise distenqjer virus, Ebola virus (including subtypes Zaire, Sudan, and Reston), Eqiune morbillivirus, &ifectious hematopoietic necrosis vinis (fish). Influenza viruses of swine, horses, seals, and fow], Kotonkan virus, Lymphocytic choriomeningiiis virus , Marburg virus, Nairobi sheep disease ^iruS; Newcastle disease virus (fowl), Obodhiang virus, Pesce-dcs-petits-ruiiiinants virus (sheep and goats). Pneumonia vims of mice, Pocine rubulavirus Oa-Piedad-Michoacan-Mexico virus). Rabies virus, Rifi Valley fever virus, Rinderpest virus, Simian parainfluenza virus 10, and Vesicular stomatitis viruses. Specific double-stranded RNA viruses which can be treated, prevented or managed by the methods of the present invention include, but are not limited to Colorado tick fever virus, Reoviruses 1-3, Orungo virus, Kemerovo vims. Rotavirus groups A-F, Eyach virus, Ibaraki virus, Golden shiner virus, chub reovirus, African horsesickness viruses 1 -9, Epizootic hemorhagic disease viruses (deer). Infectious bursal disease ■virus (fowi). Infectious pancreatic necrosis virus (fish). Human rotaviruses, and Reoviruses 1 -3, In one embodiment, the invention encompasses the treatment, prevention or management of viruses that cause, lead to or are involved in cancer. Further, the invention encompasses the treatment, prevention or management of viral strains thai are resistant to or exhibit resistance to conventional antiviral therapy. In a particular embodiment, the preferred method involves the use of variants of dextran sulfate against hepatitis B, HIV-1, HIV-2, HCMV, iVfCMV, VZV, SBV, Measles virus, Punto Tort) a, VEE, West Nile Viras, Vaccinia, Cowpox, Adenovirus Type l.HPIV, Human metapneumovirus Haemorthagic septiaeuria virus paramifluence type 3 pichiude and rhinovirus. in a specific embodiment of the invention, the vims to be treated is not a herpes virus, or more specifically, the viruses to be treated are not HSV-1 orHSV-2. Further, in another alternative embodiment, the virus to be treated is not a retrovirus, or more specifically the virus to be treated are not HIV-), HIV-2 or HTLV. The methods of the present invention are particularly well suited for human patients. In particular, the methods and doses of the present invention can be useful for immunocompromised patients including, but not limited to cancer patients, HIV infected The protocols and compositions of the invention are preferably tested in vitro, and then in vivo, for the desired therapeutic or prophylactic activity, prior to use in humans. For example in vitro assay, which can be used to determine whether administration of a specific therapeutic protocol is indicated, include in vitro cell culture assays in which cells that arc susceptible to infection with the microbe to be treated, prevented, or managed (_e.g. primary cells, transformed eell lines, patient tissue sample etc) or growth medium on which the microbe to he treated, prevented, or managed can grow (e.g., LB broth/agar, VT brotfa/agar, blood agar, etc.) are exposed to or otberwise administered a compoimd of the invention the effect oftbe compound upon the ability of the microble to grow is assessed. Compounds for use in methods of the invention can be tested in siiitable anima] model systems prior to testing in humans, including but not limited to in rats, mice, chicken, cows, monkeys, rabbits, hangers, etc. The compounds can then be used in the appropriate clinical trials. The magnitude of a prophylactic or therapeutic dose of a sulfated polysaccharide of the invention or a pharmactically iy acceptable salt, sovate hydrate, or stereoisomer thereof in the acute or chronic management of an infection or condition will vary with the nature and severity of the infection, and the route by -which the active ingredient is administered. The dose, and perhaps the dose frequency, will also vary according to the infection to be treated, the age, body weight, and response of the individual patient. Suitable dosing regimens can be readily selected by those skilled in the art with doc consideration of such factors. In one embodiment, the dose administered depends from the specific compound to be used, and the weight and condition of the patient. In general, the dose per day is in the range of from about 0.001 to 500 mg/kg, preferably about 0.01 to 200 mg/kg, more preferably about 0.005 to l00mg/kg. For treatment of humans infected by viruses, about 0.1 mg to about 15 g per day is administered in about one to four divisions a day. Additionally, the recommended daily dose ran can be administered in cycles as smgle agents or in comfamation with other therapeutic agents. In one embodiment, the daily dose is administered in a single dose or in equally divided doses. Diffcrent therapeutically effective amounts maybe applicable for different infections, as will bereadily known by those of ordinary skill in the art Similarly, amounts sufficient to treat or prevent such infections, but insufficient to cause, or sufficient to reduce, adverse effects associated with conventional therapies are also encompassed by the above described dosage amounts and dose frequency schedules. chloride), quiolones and analogs thereof (e,g., cinoxacin,, chinafloxacin, flumequine, and grepagloxacin), sulfonamides (e.g., acetyl sulfamethoxypyrazine, benzylsulfamide. to amphotericin B, itraconazole, ketoconazolc, fluconazole, intrathecal, flucytosine, miconazole, butoconazole, clotrimazole, nystatin, terconazole, tioconazole, ciclopirox, econazole, haloprogrirt, naftifinc, tcrbinafine, undecylenate, and griseofuldin. malononitrilaamindes {e-g., leflunamide), T cell receptor modulators, and cytokine receptor colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), prolactin, and intetferon (IFN), e.g.. IFN-alpha. and IFN-gamtna). treatment with another therapeutic agent, particularly an antivira] agent In one embodiment the method of the invention comprises the sdministration of one or more sulfated polysaccharides of the invention without an additional therapeutic agent. In a specific embodiment, the methods of inventioni comprise the administration of one or more sulfated polysaccharides of the invention without a fibroblast growth inhibitor. 5.2 PERIODATE TREATED AND CO-CHARGED ANIONIC POLYSACCHABIDES Tbe invention encompasses sulfeted polysaccharides that have been manipulated to reduce endocytosis by ceil receptors and to increase the flexibility of the polysaccharide backbone to enable the efficient presentation of aniomic charged groups to interact with regions on the targeted microbes. One manipulation encompassed by the present invention is the treatment of sulfeted polysaccharides with periodate. Periodate-treated anionic polysaccharides have increased flexibility due to periodate oxidation of some or all sugar residues. This treannent allows increased freedom of rotation and confoimational flexibility of the polymers and provide flexible joints to facilitate biological interactions, Periodate-treated sulfated polysaccharides of the invention can have any cottnten'on to ensure soiubiJity including, but not limited to sodium, calcium, quaternary ammonium, and potassium. Materials which may be periodate treated and used within the methods and compositions described herein also include the polysaccharides of Table 1 below. Other variations include the incorporation of non-sulfate groups, such ss carboxyroethyi groups and sulfonate groups. By lowering the degree of substitution of charge on the polysaccharide with either sulfonate or carboxymethyl groups, the ability of the polysaccharide to be endocyciosed by high charge reactor is greatly reduced, therefore increasing its plasma stability. Carboxymethyl dextran sulfate can be prepared using a modification of methods of preparation employed by others {McLaughlin and Hirbst, Con./.iees.28B: 731-736,1950; Brovm et al Arktv Kemi 22:189-206 1964). Approximately 20g of dextran is slurried in a mixture of iBopnqjanol (35Dml} and 3.85M NaOH (40ml>aDd is stirred for five minutes at 5°C in a blender. Sodium chloroacetate (t 8g) is added, and the whole mixture is stirred for 60 minutes at S^C under a nitro^n atniospbere, the mixture is removed fiom the blender and stored at 25"C for three days. The degree of carboxymethyl substitution can be adjusted by varying the time at 25'C from I day to 3 days as well as varying the mole ratio of CICHiCOONa to anhydrogjncose from Listed in Table 1 below are examples ofsuifated polysaccharides (not including dcxtran sulfate) whose anti-nucrobial activity has been demonstrated in vitro, but which previously have not been shown to have anti-microbial activity in vivo at a dosage below the cytoloxicity level of these compounds. oligosaccbaride); Sulfated bacterial glycosaminooglycan; Sulfated ctodecyl iaimnari-oligomer (alkyl obgosaccharide); Sulfated gangliosides; Sulfated laminara-oligosaccharide See, e.g.. Remington's Pharmaceutical Sciences, 8th ed,. Mack Publishing, Easton pA (J99D), Examples ofdosage forms includes, but are not limited to; tablets; caplets; An anhydrous pharmaceutical composition should be prepared and stored such that its anthydrous nature is maintained. Accordingly, anhydrous compositions are preferably Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms, in which case solid excipients are employed. If desired, tablets can be coated by standard aqueous monaqueous techniques. Such dosage forms can be prepared by any of themethods of pharmacy. In generai, phannaceuticflt compositions and dosage foims are prepared by uniformly and intimately admixing the active ingredients with liquid caniers, finely divided solid carriers, or both, and then shaping the product into the desired presentation if necessary. For exanaple, a tablet can be prepared by compression or molding. Compressed tablets can be prepared by compressing in a suitable machine the active ingredients in a free-flowing form such as powder or granules, optionally mixed with an excipient. Molded tablete can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid dilucat Examples of excipients that can be used in oral dosage forme of the invention include, but are not limited to, binders, fillers, disintegrants, and lubricants. Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, com starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid_ other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymcthyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pynolidone, methyl cellulose, pre-gelatinized starch, hydroxypropy! methyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystalline ceilulose, and mixtures thereof Exaitiples of fillers suitable for use in the pharmaceutical compositions and dosage forms diKclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystailine cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof The binder or finer in pharmaceuticai compositions ofthe invention is typically present in from about 50 to about 99 wejghtpercent of the pharmaceutical composition or dosage form. Suitable forms of microcrytalline cellulose include, but are not limited to, the materials sold as AVICE1^PH401, AVlCEL-PH-103 AVICELRC-581. AVICEL-PH-lOS (available from FMC Corporafion, AmericHn Viscose Division, Avicel Sales. Marcus Hook, PA), and mixtures thereof An specific binder is a mixture of microcrystalline cellulose and sodium carboxymethod cellulose sold as AVICEL RC-581. Suitable anhydrous or low moisture excipients or additives includes AVICEl^PH-1 OS'™* and Starch 15O0 LM. Disintegrants are used in the composition of the invention to provide tablets that disintegrate when exposed to an aqueous environment Tablets that contain too much disniegrant may disintegrate in storage, while those thai contain too littie may not . disintegrste gt a desired rate or under the desired conditions. Thus, a sufficient amount of disintegrant that is desired too mock nor too little to detrimentally alter the release of the active ingredients should be used to form solid oral dosage forms of the invention. The amount of disintegrant used varies based upon the type of formulation, and is readily discemibletothoseof ordinary skill in the art. Typical phaimaceutical compositiong comprise from about 0-5 to about 15 weight percent of disinlegrant, specifically from about 1 to about 5 weight percent of disintegrant. Disintegrant that can be used in pharameceutical compositions and dosage fonns of the invention include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscannellose sodium, crospovidonc, polacrilin potaasinm, sodium starch glycolate, potato or tapioca starch, pre-gelatinized starch, other starches, clays, other aigins, other celluloses, gums and mixtures thereof. Lubricants that can he used in pharamaceutical compositions and dosage forms of the invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, com oil, and soybean oil), zinc stearate, ethyl oleatc, ethy] laureate, agar, and mixtures thereof Additional lubricants include for example, a syloid sihca gei (AEROSIL 200, manufactured by W.R, Grace Co. of Baltimore, MD), a coagulated aerosoi of synthetic silica (marketed by Degussa Co. of Piano, TX), CAB-O-SJL (a pyrogenic silicon dioxide product sold by Cabot Co. of Boston, MA), and mixtures thereof. If used at all, lubricants are typically used in an amount of less than about 1 weight percent of the pharmaceutical compositions or dosage forms into which they are incorporated. 5A.1 Delayed Release Dosage Forms Active ingredients of the invention can be administered by controlled release means or by delivery devices that are well known to those of ordinary skill in the art. Examples include, but are not limited to, those described in U.S. Patent Nos.: 3,84.5,770; 3,916,899; 3.536,809; 3,598,123; and 4,008,719, 5,674.533, 5,059,595, 5,591,767, 5,120,548. 5,073,543.5,639,476, 5,354,556, and 5,733,566, each of which is incorporated berem by refaremce. .Such dosage form.s can be used to provide slow or cootrolled-release of one or more active ingredients using, for example, hydropropylmetbyt cellulose, other polymer matrices, gels. penneable membranes, oamotic systems, multilayer coatings, microparticles, but are not limited to, solutions ready for inj ection, dry and/or lyophylized products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection (reconstitutable powders), suspensions ready for injection, and emulsions. acetone; various alcohols such as ethanol, oleyl and tetrahydrofuryl; alky! sulfoxides such as dimethyl sulfoxide; dimethyl acetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such as polyvinylpyirolidone; Kollidon grades (Povidone, Polyvidone); urea; and various water-soluble or insoluble sugar esters such as Tween SO (polysozbate 80) and Span 60 (sorbitan raonostearale). SA.6 Mucosal Dosage Forms Mucosal dosage forms of the invention include, but are not limited to, ophthalmic solutions, sprays and aerosols, or otherforms known to one of skill in the art. See, e.g-.. Remington's Pharmaceutical Sciences, l8theds., Mack Publishing, Easton PA (1990); and Introduction to Pharmaceutical Dosags Forms, 4tli ed., I^a & Febiger, Pbiladeiphia (1985). Dosage forms suitable for treating mucosal tissues within the oral cavity can be fbnnulated as mouthwashes or as orai gels. In one embodiment, the aerosol comprises a carrier. In anothea: embodiment, the aerosol is carrier free. The sulfated polysaccharides of the mvention may also be administered directly to the lung by inhalation. For administration by inhaJanon, a sulfeled polysaccharide can be conveniently dehvcred to the lung by a number of different devices. For example, a Metered Dose Inhaler ("MDF') -which utilizes canisters that contain a suitable low boiling pTOpellant, e.g., dichlorodiQuoromethaae, trichlorofluoromethane, dichJorotelrafluoroethane, carbon dioxide or other suitable gas can be used to deliver a suifeted polysaccharide directly to the Jung. MDI devices are available from a ohmber of stqjpUers such as 3M Corporation, Aventis, Bnehiinger Ingieheim, Forest Laboratories, Glaxo-Wellcome, Schering Plough and Vectura. -Mlematively, a Dry Powder Inhaler (DPI) device can be used to administer a sulfated polysaccharide to the lung {see, e.g., Raleigh et al., Prac. Amer. Assoc. Cancer Research Annual Meeting, 1999, 40, 397, which is herein incorporated by reference), DPI devices typically use a mechanism such as a burst of gas to create a cloud of dry powder inside a container, which can then be inhaled by the patient. DPI devices are also well known in the art and can be purchased fom a number of vendors which include, for gxample, Fisons, Glaxo-Wellcome, Inhale Therapeutic Systems, ML Laoratories, Qdose and Vectura. A popular variation is the multiple dose DPI ("MODPr") system, which allows for the delivery of more than one therapeitic dose. MDDPI devices are available from companies such as AstraZeneca, Glaxo Wellcome, WAX, Schering Plough, SkyePharma and Vectura, For example, capsules and cartridges of gelatin for use in an liquid such as alcoiioi, water, polyethylene glycol or a perfluorocarfcon. OptionaUy, another material may be added to alter the aerosol properties of the solution or suspension of sulfated polysaccharide. Preferably this material is liquid such an an alcohol, glycol, polyglycol or a fatty acid. Other methods of formulating liquid drug solutions or skilled in the phannaceutical arts, and depend of the particular site or method which a given pharmaceutical composition or dosage form will be administered. With that feet in mind, typical excipients include, but are not limited to, were, ethanol ethylene glycol, propyjene glycol, butane-l,3-d2ol, isopropyl myristate, isopropyl palmitate, oiinerai oil, and mixtures thereof, which are non-toxic and pharmaceutically acceptable. Examples of such additional ingredients are well known in the art. See, e.g., Rcinington's Phannaceutical ScienceIStii eds., Mack Publishing,Easton PA (1990). The pH of a pharmaceutical composition or dosage form, or ofthe tissue to which the phsnnacetitical composition or dosage form is applied, can also be adjusted to improve delivery of one or more active ingredients. Similarly, the polarity of a solvent carrier, its ionic strength, or tonicity can be adjusted to improve delivery. Compounds such as stearates can also be added lo pharmaceutical compositions or dosage forms to advantageously alter the hydrophilicity or lipophilicity of one or more active ingredients so as to improve delivery. In this regard, stearates can serve as a lipid vehicle for the formulation, as an emulsifying agent or surfactant, and as a delivery-enhancing or penetration-enhancing agent. Different salts, hydrates or solvates of the active ingredients can be used to further adjust the properties of the resulting composition, S.4.7 Condoms and Prophylactic Devices In a preferred embodiment of the invention, the sulfated polysaccharides of the invention can be used as a coaling for a condom or other prophylactic device. Similarly, the sulfated polysaccharide can be used as a coating for surgical instruments and protective devices including, but not limited to rubber gloves, surgical masks, CPR aids, tongue depressors, bandages, sponages, napkins, dental devices and thermometer be covers. When a sulfated polysaccharide of the invention is used as a coating as described herein, it is preferred to have a molecular weight higher than 500,000. ID a preferred embodiment, a sulfated polysaccharide of the invention is combined with a talcum lubricant powder used to line surgical gloves. The methods of using the sulfated polysaccharides of the invention as a coating will be well known by the skilled artisan. Similar methods can be found in United Stales Patent No. 4,869,270 which is incorporated herein by reference. S.4.S Disinfectants and Detergent in one embodiment of the invention, the sulfated polysaccharides of the invention permissive cell line (e.g. primary cells, transformed cell lines, patient tissue samples, etc) or growth medium (e.g., LB broth/agar, YT broth/agar, blood agar, etc). The growth/infection of the microbe can be compared the growth/infection of the microbe in the absenseofthe compound of the invention. Anti-microbe activity of the compound of the invention is demonstrated by a decrease in microbe growih/infection in the prcsence of the compoundof BUSA; for discussion regardingELlSAsJee,e.^.,Ansube] etal, fids, 1994, Current Protocols in Molecular Biology, Vol. i, John Wiley & Sons, Inc., New York at 11.2.1), In another embodiment the microble is a figure. Standard models of in vivo antifungal activity includes, but are not limited to, those described in The Handbook of a rabbit model of bacterial osteomyelitis of the tibia; a rai tnodej of arthroplasty; a rabbit model of arthroplasty, a mouse model of streptococcal fasciitis; a rabbit model of bacterial endocarditis; an adult rat mode) of meningitis; and a rabbit tnodel of bacterial meningilis. In one specific embodiement, the growth rate of a microbe in a subject can be estimated by fee titer: of antibodies against ike microbe in the subject Antibody senim titer cao be determined by any metbod well-known in the art, for example, but not limited to, the amount of antibody or antibody fragment in serum samples can be quantitated by, e.g., ELlSA. Additionally, in vivo activity of a sulfated polysaccharide ciaa be determined by directly admimstering the compound to a test animal, collecting biological fluids (e.g., nasal aspirate, throat swab, spuhrni, bioncho-alveolar lavage, urine. Saliva, blood, or serum) and test&ig the fluid for anti-microbial activity. In embodiments where samples to be assayed for microbial leveis are biological fluida/'clinical sanapies (e.g., nasal aspirate, throat swab, sputum, brodcho-alvcolar lavage, urioc, saliva, blood, or serum), the samples may or may not contain in tact cells. Samples ftoin subjects containing inlact cells can be directly processed, whereas isoistes without intacttellsmay or maynotbe first cultured on a permissive cell line(e.^. primary cells, transformed ccU lines, patient tissue samples, etc) or growth medium (e.g)., LB broth/agar, YTbroth'agar, blood agar, etc.). Cell suspensions can be cleared by centriugation at,(eg), 300xg for 5 minutes at room temperature, followed by a PBS, pH 7-4 (Ca++ and Mg++ free) wash under the same conditions. Cell pellets can be resuspended in a small volume of PBS for analyas. Primary clinical isolates containing intact cells can be mixed with PBS and centrifiiged ai 300xg for 5 minutes at room temperalnre. Mucus is removed frotn the interlace with a sterile pipette tip and cell pellets can be washed once more with PBS under the same conditions. Pellets can then be resuspended in a small volume of PBS for analysis. In another embodiment, a compound of the invention is administered to a human subject infected with a microbe. The incidence, severity, length, viral load, mortality rate of infection, etc. can be compared to the incidence, severity, length, viral load, mortality rate of infection, etc. observed in human subjects infected with a microbe in the absence of a compound of die invention or in the presence of a placebo. Anti-microbial activity of Ihc compound of the invention is demonstrated by a decrease in incidence, severity, length, viral load, mortality rate of infection, etc. in the presence of the compound of the invention. Any method known in the art can be used to determine anti-microbial activity in a subject such as those described previously. Additionally, in vivo activity of a sulfated polysaccharide can be detemiined by directly administeriiig the compound to an animal or human subject, collecting biological fluids/clinical samples e-g-. nasal aspirate, throat swab, sputum, broacho-aiveolar lavage, urine, saliva, blood, or serum) and testing the biological fluids/clinical samples for anti-vtTal activity {eg., by addition to cells m culture in the presence of the microbe). In general, m vivo stability can be determined by a variety of models known to the skilled artisan. In panicular, in vivo stability can be determined by a kidney perfusion assay. For either type of analysis, the test compound may be labeled, for example with tritium. A kidney perfusion technique is described in detail in Tay el ai. {Am. J. Physiol., (1991), 260: F549-F554). Briefly, rat kidneys. e.g., from male Sprague-Dawiey rats, are perfused with 5% bovine serum wnin (BSA) in modified Krebs Henseleit buffer containing amino acids and continually gassed with 95% 0% -5% CO1. Samples that have been perfused may be subjected TO ion-exchange chromatography using, for example, a 19x 1/cm2 column of sepharose Q. Samples are applied to the column in 6 M urea, 0.05 M Tris, 0.005% (w/v) Chaps, pH 7-0, and eluted width a linear gradient of 0.15-2.5 M NaCi in the same buffer at a flow rate of 0.5ml/minute. Recoveries using this technique are very good- The foregoing has demonstrated the pertinent and important features of the present invention One of skill in the art will be appreciate that numerous modifcatfons and embodiments may be devised. Therefore, it is intended that the appended claims cover all such modifications and embodiments. 6. WORKING EXAMPLES The following examples are for the purpose of illustration only and arc nor intended as limiting the scope of lie invention. 6.1 Example 1: Synthesis of a sulfated dextran having a salfatiOQ of 9.5% Dextran T20(Bverage molecular weight 20.000) was dried in vacuo at 60°C overnight The dried compound flOO g) was dissolved in 640 ml formamide (FA). Chlorosuifonic acid (CSA) SO ml was added to FA 200 mi at a maximum of 45°C in a 3-necked Bask, then cooled in ice-water. The amount of CSA determines the ultirnate sulfation of the sulfated dextran (1 SO ml CSA to 200 ml FA yields approximately 17% sulfur). The CSA/FA mix was slowly added {over two hours) to the dextran at a temperature of 40'C- After all of fccCSA/fA was added, the mixture was stirred for 15 minutes at a temperature of 45°C, The mixture was cooled to 25'C and 28% NaOH was added slowly to give a pH7.S-8.5 with a maximum temperature of 50°C. For the-first precipitation, 3 L of ethano) were added with stirring. Stimng was stopped and the mixture Was allowed to stasd. The supematant was decanted and the precipitate was redissolved in 1.5 L of water, For the second prccipitation 1.5 L ethano were added with stifling and then the mixture was allowed to stand far ca-o hours. The supermatant was decanted and the precipiate was redissolved in 900 ml of water, to which 17 g NaC! was added. For the third precipitation 800 ml ethanol were added -with stirring and the mixture Was allowed to stand for two hours. The optical rotation maximum was measured. The supermatant was decanted and the precipitate was redissolved in 500 ml water. 2.8 g Na2HPO4 and 2.6 g NaH2POA were added. For the final precipitation 5 L ethanol were added and the precipitate Was filtered on a glass filter end dried in vacuo ai 50°C. 6.2 Example 2; Periodate Oxidation Following the modified method of Smith degradation used by Sandy JD, Siochem J., 177: 569-574, 1979; chrondroitin sulfate (240 mg) was dissolved in 0.25M NaC10« (47 ml) at room temperature. 5 ml of 0.5 MNalO4 was added and KOH was used to adjust the mixture to pH 5, The reaction was allowed to proceed in the dark for 72 hours. The mixture was then dialysed in visking tubing to remove the periodate, 63 Examples: Introduction of Anionic Sutfur Groups to Carboxymelhy! Dextran Sulfated form of carboxymethyl dextran (average mw 20,000) with a sulfur content of 9,5%. Carboxymethyl dextran (CMD) is dried in vacuo at 60°C overnight. CMD ("100 g) is dissolved in 640 ml formamide (FA). Chlorosulfonic acid (CSA) 80 ml is added to FA 200 ml at maximum of 45°C in a 3-necked flask then cooled in ice-waiet. The amount of CSA will detemiine the ultimate sulfur content of CMD (180 ml OSA to 200 ml FA yields approx 17% sulfur). The CSA/FA mix is added slowly (over 2 hours) to CMD at a temperature of 40'C. After all is added the mixture is stirred for 15 minutes at a temperature of 45°C- The mixture is cooled to 25°C and 28% NaOH is added slowly to give a pH 7.5-S.5 with a maximum temperature of 50C. For the first precipitation, 3 L of ethanol is added with stirring. Supernatant is decanted and then residue is redissolved in i.5 Lofwater. For the second precipitation 1.5 L ethanol added with stirring and then allowed to stand for 2 hours, Superoatant is decanted and residue is redissolved in 9O0 ml of water and then added to 17 gNaCl- For the third precipitation 800 ml ethanol is add with stirring and allowed fo starsd for 2 hours. The optical rotation maximum should be 0.3. Supernatant is decanted and the residue is redissolved in 500 m water. Add 2.8 g"Na:HPO4 and 2.6 g NaH2PO*. For the final precipitation 5 L ethanol is added and filtered coi a glass filter and is dried in vacuo at 50°C. Sulfonated form of carboxymethyl dextran (average molecular weight 20,008). Step 1. Dissolves g dextran in water. Add l00mgborohydride stir at room temp, for 30 min. Step 2, Add sodium hydroxide pellets (l0g) and stir until dissolved and then sulfonatc{12g). Step 3. Heat at 70°C for 7 h. After 3 hours add a further 3 g of sulphonate. Continue heating for 4 hours. Step 4. Neutralise with 5M HCl to pH 7.5 (Total volume(T) = 75ml) and gradually add 200 ml ethanol with good stirring. Stop stirrer and stand i hour. Step 5. Decant supernatant; redissolve in water (T = 60 ml) and add 150 ml ethanol with good stining. Stand ] hour. Step 6. Repeat as Step 5. Step 7. Decant off the supernatant- redissoive the residue in 60 ml water and ppte in 600 ml ethanol. Some concentrated sodium chloride solution may be added to tiie mixture to aid precipitation. Step 8. Filter and dry in vacuo. Yield approx. 6 g. 6.4 Example 4: In vivo anti-viral activity The in VIVO anti-viral activity of dextran. sulfate and variants of sulfeted dextrans was assessed in a pharamacokinetic study involving single intravenous doses of 60 mg/kg commercially available (~I7% sulfor) dextran sulfate (DS) of 40,000 mw (groupi); DS I 500,000 mw (group 2); dextran sulfate (12.2% sulfete)(DES6) 40,000 mw (group 5); DES6 500,000 mw (group 4) given to three male and three female rats and a multi-day injection of 60 tng/kg DES6, 500,000 mw given to an additional group of three rats (group 5). Rats were Sprague-Dawley, previously cannolatcd in the vena cava. Blood was drawn at various tinies after iajectioo and assessed for and-HlV activity in an acute infectivity cytoprotcction assay system utilizing HTV-1 RF virus with CEN-SS cells using fee MTS staining mettiod forccO viability (based on Witvrouw er a/„ J. Acqur. Immun.Def. Syndr., 3:343-347, 1990). The results shown in Figure 2 indicate that DS was, as expected, highiy toxic at ^le&e doses with only one lat surviving beyond 24 hours. In contrast, good survival and circulattag anti-HTV activity for as long as ]20iiouiBafler injection were observed in Che DES6 Ireat&d rats. Figure 2 represents summary daia from the five groups of animais. Each data point Ttyresents the concentration of circulatiEg antiwrai aclivily at tiroes after injectioa. CoDcentration was calculated by determintcig the IC50 of confound in the blood. As can be calculated from the raw data. DES6 of both rnolecular weights showed a prolonged half-life in the biood of between 12 and J S hours, and an extended and-viral activity (circulating concentration above the ICso) beyond 72 hours. With three repeated injectjons of group 5 animals a steady state concentratiQn was reached. Results are expressed in Figure 2. The data indicate that any mortality associated with DES5 was probably due to complications associated wn'th the cannulatjon, since the MTD in noo-cannulated animals is >850nig/kg. 6.5 Example 5: Effect on Pro-thrombin/Tlirombin and Activated Partial Thromboplastin Time As noted above, inhibition of coagulation has been a repeatedly observed side effect of sulfated polysaccharide treatment, particularly with conventional dextran sulfate treatment. The purpose of this study was to evaluate the effects of DES6 compared to commercially available DS on prothrombin time (PT) and activated partial thromboplastin time (aPTT)- All ^)ecimeDs were "spiked" wifli the test compound prior to submission to a CUnical Pathology Laboratory, The specimens were delivered along with reconstituted human plasma purchased from Sigma, Immediately prior to analysis 600 ^1 of tiie Sigma homan plasma was added to each spechnciL A Bio-Mcrieiw Coag-A-Male MTX II Analyzer was used to measure Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT)- The PT reageni: used was Simplastin L and the APTT reagent used was Platelin L; all reagCTits were obtained from Maximum Tolerated Dose The multiple toxicity dose (MTD) of DES6 was assessed in & series of experiments where groups of five rats were given 100 or 200 mg/kg doses of DES6 mw= 500,000. Body weight and overall behavioral assessments were determined for five days after injection. There wens no overt signs of toxicity as determined by observation and "body weight measurements. Subsequently rats were given a 500 mg/kg injection and observed for a further five days also without signs of toxicity. Finally animals were given a dose of 850 mg/kg. Results are provided below in Table 4- 6,6 Example 6: In vitro anti-viral assessment of sulfated polysaccharides The studies included assessment of five test compounds at a high test concentration of 5O0 ng/ml in human peripheral blood mononuclear cells (PSMCs)- Methods All test compounds #3 (dextran sulfate 17-20%), #4 (sulfated dextran, 9.5% sulfiu-, molecular weight 30.000), and #6 (sulfated dexhan, 12.2% sulfur, molecular weight 36,000) were aolubilized in H2O at 40 mg/ml. The compounds were visually completely soluble and colorless. Compound were light protected and assays were performed in a maimer which minimized incidental light. Compounds were stored at -20°C following solvation- Viruses The low passage pediatric isolate RoJo was derived in fee laboratories of Southern Research Institute. RoJo is a presumed subtype B virus. PSMC Isolation and Blasting Peripheral blood monocular cells CPBMCs) were obtained from normal hepatitis and HlV-1 negative donors by ficoll hypaque gradient separation. The mononuclear cells were washed to remove residual gqsaration media, counted, viability determined and resuspended in RPM3 1640 medium supplemented with 15% FBS (heat inactivated), 2 mM L-ghitamine, l00U/mLpenicillin, 100 μ g/mL streptoycin, and 10 μ g/mL gentamycin with 2 μ gi'mL phytohemagluttin (PHA) at 1 X 10 cells/mL, The cells were cultured for 48 to 72 h at 37°C, 5% CO;. Following incubation, cells were collected by centrifiiguation, washed and resuspended in RPMI1640 supplemented with 15%KBS (heaiinactivaied), 2 mML-glutamine, 100 U/mL peniciljin, 100 fig/mL streptomycin, and 10 μ g/mL gentamycin with 20 U/mL recombinant 11^- R&D Systems, Minneapolis, MN). IL-2 was included in the culture medium to maintain the cell division initiated by the PHA mitogenic stimulation. The cultures were then maintained until use by 1/2 culture volume change with fresh 11^2 containing medium every three days. PBMCAssay Human peripheral blood mononuclear cells from a minimum of two donors, that have been blasted with PHA and IL-2, were counted, viability determined by Trypan Blue dye exclusion and mixed in equal ratios. Pooled donors were used to minimize the variability observed between individual donors which results from quantitative and qualitative differences in HTV infection and overall response to the PHA and IL-2 of primary lymphocuyte populations. The cells were resuspended at 1 x 106 cells AnL in RPMl 1640 without pheno! red supplemented with 15% Fetal Bovine Serum (heat inactivated), 2 mM L-glutamine, l00U/mLpenicillin, l00/tg/mLstreptoinycin, 10 ;μ g/mL gentamycin and 11^2 (20 U/mL, R&D Systans, Minneapolis, MN). Fifty microliters of cells were then distributed to the inner 60 wells of a 96 well round bottom microtiter culture plate in a standard format developed by the Infectious Disease Research department of Southern Research Institute. Each piate contains cell control wells (cells only), virus control wells (cells plus virus), and experimental wells (drug plus cells plus virus). Serially dihited compounds were added to the microtiter plate followed by the appropriate prehtered dilution of HIV- I RoJo, ,All samples were assayed in triplicate with a replicate plate without vims for the determination of compound toxicity. The final volume per well was 200 μ L. The assay was incubated for 6 days in a humidified atmosphere at 37C, 5% CO2, after which supermatants were collected, for analysis of RT activity and sister plates analyzed for cell viability by MTS dye reduction- Wells were also examined microscopically and any abnormalities noted. MTS Staining for Cell Viability At assay termination the assay plates were stained with the soluable tetrazolium-based dye MTS (CellTiter96® Reagent Promega) to determine cell viability and quantify compound toxicity. MTS is metabolized by the mitochondria enzymes of metabolically active cells to a soluable formazan product allowing the rapid quantitative analysis cell viability and compound cytotoxicity. This reagent is a single stable solution that does not require preparation before use. At termination of the assay 20 μ L of MTS reagent was added per well and incubated for 4 h at °C. Adhesive plate sealers were used in place of the lids, the sealed plate was inverted several times to mix the soluble forman product and the plate was read spectrophotmnetrically at 490 am with a Molecular Devices Vmax plate reader. Reverse Transcriptase Assay for Culture Supernatants Reverse transcriptase (RT) activity was measured in cell-free supernatants. Tritiatcd thymidine triphosphate (NEN) (TTP) was resuspended in distilled HzO at 5 C/mL. Poly rA and oligo dT were prepared as a stock solution which was kept at -20°C. The RT reaction buffer was prepared fresh on daily basis and consists of 125 pi-1.0 MEGTA, 125 ^ dH20, 110μ L 10% SDS, 50μ LOMTris(pH 7.4), 50μ A.OM DTT, and40μ L 1.0M MgCb. These three solutions were mixed together in aratio of two parts TTP, one part poly rAjoligo dT, and one part reaction buffer. Ten microliters of this reaction mixture were placed in a round bottom microtiter plate and 15 μ l of virus containing supermatantt was added and mixed. The plate was incubated at 37°C in a water bath with a solid support to prevent submersion of the plate and incubated for 60 minutes- Following reaction, the reaction volume was spotted onto pieces of DE81 paper, washed 5 times ibr 5 minutes each in a 5% sodium phosphate buffer, two times for one minute each in distilled water, two times for one minute each in 70% ethanol, and then dried. Opti-Fluor O was added to each — ^^,.—^ . ^-1 .. . , _., — . ^ . —^ _ ,— - . _^ — _x— --—- ^—^^ • ^ ' Table 5 compares the previous and current antivirai evaluations in PBMCs. The previously identified ICM and antiviral efficacy of DK 17-20% Sulfation was verified with an IC5o of 0.5 μ g/IQl m Ihese Experiments. This is within the standard 3-fo)d error predicted for the PBMC assay. In addition, the second experiment demonstrated that compound #3 is non-cytotoxic to PBMCs at 500 (μ g/ml. In this set of evaluation the initial antiviral assessments of DES 9.5% Sulfation and DES 12.5% Sulfation were performed Both compounds were non-cytotoxic at 500 /ig/ml and 50% iohihitory concentrations were derived. DES 12.5% Sulfetion displayed antiviral activity equivalent to DES 17-20% Sulfclion based upon the calculated ICjo, 1.6 vs. 0.5 μ g/ml, respectively. Additionally, examination of the antivirai efficacy curves suggests diat the 2 compounds are of equal potency. In contrast, DES 9.5% Sulfation v/as 39-fold less active than DES 17-20% Sulfation and 12-foid less active Than DES 12.5% Sulfation. 6.7 Example 7: In vitro anti-viral assessment of sulfated polysaccharides Tte following compound have been tested for in vitro anti viral activity. Sample 3(dextran sulfate, I7 -20% sulfur, molecular weight 39,700), sample 4 (dextran sulfate, 9.5% sulfur, moiecular weight 30,000) and sample 6 {dextran sulfate, 12.2% sulfur, molecular weight 36,000). All three compounds exhibited significant anti-viral activity . against HIV-l RoJo virus. DES 9.5% Sulfation and DES 12.5% Sulfation were also assessed against a range of HIV-I clinical isolates, including subtype representative isolates, SIV and HTV-Z The inhibition of HTV-IADA and BaL replication in monocyte/macrophages was also assessed, DES 9.5% Sulfation and DES 12.5% Sulfation were prepared as described above in Example 4. Mutations in bold face type in Table 7 represent key resistance mutations in the indicated genes. PBMC Isolation and Blasting Peripheral blood monocular cells (PBMCs) were obtained as described in Example 6. PSMCAssay PBMC assays were carried out as described in Example 6. Monocyte isolation, culture and infection Peripheral blood monocytes were isolated from normal HIV-) negative donors by plastic adherence following ficol hypaqne purification of the baffy coat, as described above for PBMCs. In many cases the same donor used to produce the PBMC populations was also used to produce monocyte/macrophages, however unlike PBMC population monocyte/macrophage, donors were never pooled. Following a two hour adliereDce in RPMI1640 without phenol red supplemenf ed with 10% human pooled AB serum (heat inactivated), 2 mM L-glutamine, 100 U/mL penicillin. 100 μ g/mL streptomycin, 10 μ g/mL gentaraycin, cultures were washed to remove non-adherem cells. The monocytes were released from the plastic by vigorous pipetting with Ca^2 and Mg2" free PBS. Adherent cells were assessed for purity by nonspecific esterase staining (a-napthyl butyrate specific esterase. Sigma Chemical Co.), and/or liability by Trypan Blue dye exclusion, counted and resuspended in RPMI 1640 supplemented with 10% Fetal Bovine Serum (heat inactivated). 2 mM L-giutamine, 100U/mL pemciih'n, 100 μ g/mL streptomycin, 10μ g/tnl. gentamycin at 1 X 106 monocytes per ml. The monocytes (1 x 105 per 0.2 cm well) were then cultured for six days, allowing maturation of the cells to a macrophagehlike phenotypc. At day six Gie cultures were washed three times to remove any non-adherent cells and serially diluted test compounds added followed by Hie addition of a pre-litered amount of HTV-l vims, if microscopic observation of the wells demonstrated a 70% or greater confluency of tiie monocyte/macrophage monolayer. Cultures were washed a final time by media removal 24 hours post infection, fresh compound added and the cultures continued for an additional six days. The assays were preformed using a standardized microtrter plate format, which uses only the inner 60 wells of a 96 well plate for assay pmposes. The outers contain media and acts an a evaporation barrier. Each plate contains cell control weils (cells only), vims control wells (ceils phis virus), and experimental wells (drug plus cells plus virus). HIV p24 antigen contents to assess vims replication was measured at assay termination by a commercially available p24 ELISA assay (Coulter) OD cell-free supematants, and compound cytotoxicity by MTS dye reduction. AZT, HIV-1 reverse nucleoside transcriptase inhibitor and dextransuliate, an attachment inhibitor, were used as positive control compounds and ma in parallel with each determined. At termination of the assay culture plates were rercoved from -the incubator and observed microcopically. Any uniqueindings were noted. MTS staining for cell viability MTS staining was carried out as described in Example 6. Reverse Transcripiase Assay fur Culture Supematants Reverse transcriptase (R.T) activity was measured in cell-free supematants as described in Example 6. P24 Antigen ELISA ELISA kits were purchased from Coulter Electronics, The assay was performed according to the manuactures instructions. Controi curves were generated in each assay to accurately quantitate the amount of p24 antigea in each sample. Data were obtained by spectrophotometric analysis at 450 pm using a Molecular Devices Vmax plate reader. Final concentrations were calculated from the optical density values using the Molecular Devices Soft Max software package, Result ICso (50%, inhibition of virus replication), TCsj (50% reduction in cell viability) and a therapeutic index (TI, TC5o.'ICso) were calculated- The results are Eunmiarized in Table 8, The antiviral data for each test include die relevant raw data vahies from tiie triplicate tests for virus replication (RT (cpm) for PBMCs and p24 (pg/ml) for monocytes) and cell viability (OD 490) for MTS dye reduction. The IC50 and TC50 values were calculated by linear recession. The TI represents the ratio of the ICsrACso, and is used to determine relative potency between compounds. The graphical representation shows the relationship between antiviral efficacy (%VC) find compound toxicity (%CC) expressed as a percent of the control, virus no compound or ceils no compound, respectively. TABLE 8: Summary of the Range of Action Testing active against the broad range of RTV- 1 isolates tested, as weii as displaying activity against a multi-drug resistant virus , SIV and HIV-2, This, these compounds are broadly anti-retro viral. DBS 12.5% Sulfation was active against all viruses tested. It was least active against the subtype C (IC5010.3 μ g/ml) and G (IC5011,7μ g/ml) viruses. It also efficicenty inhibited tbc replication of HIV-! ADA, DBS 12.5% Sulfation also displayed good antiviral activity with a clinical isolate of HIV-2 and the SIVmacZS 1 isolate of SIV. It also displayed significant activity against the muiti-drug resistant virus isolate MDR.769. Thus, DES 12.5% Sulfation is active against a broad range of HIV-l clinical isolates, multi-drug resistant viruses and other retroviruses. DES 9.5% Sulfetion showed a heterogeneous response (variation in IC50) to the various viruses tested with activity ranging from inactive to active, DES 9.5% Sulfation has been previously demonstrated to be less active than DES 12.5% Suifetion in HTV-l RoJo infecied PBMCs, and this difference was again demonstrated here (37'foid less active). Examination of the antiviral curves for those viruses (ADA and Ba-L and the subtype G virus) for which DES 9.5% Sulfation was inactive suggests that it would be active at higher test concentrations. DES 9.5% Sulfation was also active against the MDR769 HIV-I strain (ICso 13 μ g/ml) and a clinical isolate ofHIV-2(IC5o 5.1 μ g/ml). Thus, despite lower over all potency it is still highly potent against multi-drug resistant HIV-l and HIV-2. DES 9.5% Sulfation and DES 12.5% Sulfation were tested against a range of HIV-I clinical isolated and two other retroviruses (HlV-2 and SIV-l) and found to be broadly anli-retroviral. Additionally, these results show thai the compounds are active against a resistant clinical isolate carryicg the T215 Y mutation for multi-drug resistance to R.T inhibitors. The data also demonstrated that although DES 12.5% Sulfation was more potent than DES 9.5% Sulfation on an IC50 basis, their range of IC50s on the panelof viruses were comparafaic. Thus, it is likely that both inhibit virus reT)licatiQQ via a coit^arable roechanism of action. Finally, the demonstration that the compoynds are actrve against HIV-2 and SIV-l show that they arc applicable to other retroviruses. 6.8 Biodistribution of 3 Compound of the Invention Male Sprague-Dawley rats obtained from Charles River Laboratories (Raleigh NC; ca. 377-402g) were dosed with [3H]Des6 40K by intravenous bius or oral gavage administration. Distrubtion of total critium content in plasma, lymph, and cervical lymph nodes was quantitated in samples collected at 6 or 12 hours following dosing. The study design is outlined in Table 9. Rats were divided into three treatment administration groups. Doses were formulated in phosphate buffered saline vehicle (pH= 7.4) so as to deliver them in approximate volumes of 1 .S mL/kg (iv) and 2.1 ml/kg (oral gavagc). Prior to the time of biologicsi sample collection at 6 or 12 hours ailer dosing, animals were anesthetized with kctamine/xylazine (7:1, ca. 120 mg/kg), and the thoracic lymphatic duct was cannnlated as described in Waynforth, H.B. and Fleckneil, PJV, (1992). Experimental and Surgicai Technique in the Rat, 2nd ed.. Academic Press, New York. At the time of sample analysis, blood was collected by cardiac puncture and lymph was collected via the lymphatic duct cannula. Blood was processed for plasma by centrifiigation at ca. lOOOg for 10 minutes. Cervical lyn^h nodes were collected from each animal ai times specified in Table 9. Except where noted, total radioactivity was quantitated LD duplicate by hquid scintillalion spectrometry for a3J hio!og!c;al samples collected. Results of the study are described in Table 10- In addition to listing total radiokbel content in plasma, lymph aad lymph nodes; the lympb:plasma and lymph noderplaama ratios are also provided for each animal. Overall, concentration of [3H]De6 40K associated total radioactivity were higest in animals treated by iv administration, compared with oral administration, with higest concentrations in plasma and lymph at 6 h compared with 12 h- Plasma and lymph ['HlDesfi 40K-eq concentrations at 12 h were approximately 1-2% of (hose obtained at 6 b. However, the concentrations of total radioactivity in lymph nodes were sirailar between these two time points following iv administration. Anima! G961 in Group 2, died fbiiowing anesthesia and prior to cannuladon of the thoracic duct for collection of lymph. "lymph could not be collected from this animal; however plasma and iymph nodes were harvested for quantitation of totai radioactivity. Total radioactivity in these collected biological media were found to be significantly greater than the other two rats thai survived througtout the surgery. Lymph nodes in this animal ware observed to be larger than the other two animals in Group 2. Following oral administration of ["'HIDes6 40K, concentrations of plasma total radioactivity were comparable lo those obtained at 12 h following iv administration. Mean total radioactivity in lymph following oral administration was approximately 63% of those obtained at 12 h after iv administration; while total radioactivity in lymph nodes was only approximately 0-4% of those obtained following iv administration. The lymph:plasma ratios increased in rats between the 6- and 12-h time points following iv administration (compare 0.14 and 0,54 vs. 1,7 and 1.3 for the 6- and 12-h time points, respectively), as plasma total radioactivity signiScantly decreased. The iymph/plasma ratios following oral administration were approximately one, indicating equal distributioii of total radioactivity in these two media at 12 h. The lymph node:pIasma ratios iocreased to a much greater extent at the 12-h time point compared with those obtained W 6 h. The increase over time was much grater than those obtained in lymph over the same time course. These data suggest that [^H]De86 40K associated radioactivity distributes to a large degree into lymph nodes and that the rate of elimination from this tissue is slow. In contrast, the distribution profile to lymph nodes was different following oral administiation, where the lymph node;plasma were only approximately 0.50-0.82, and demonstrate less distribution of total radioactivity into lymph nodes by the oral route. WE CLAIM: 1. A minimal sulfated polysaccharide having a percentage of sulfur above 6% and below 13% with respect to the simple sugar residue and wherein the molecular weight is above 5,000 g/mol. 2. The sulfated polysaccharide as claimed in claim 1 wherein the percent of sulfur is above 7% and below 13%. 3. The sulfated polysaccharide as claimed in claim I wherein the percent of sulkier is above 8% and below 13%. 4. The sulfated polysaccharide as claimed in claim J wherein the percent of sulfur is above 9% and below 13%. 5. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide is a co-charged anionic polysaccharide. 6. The sulfated polysaccharide as claimed in claim 5, wherein the co-charged anionic polysaccharide is co-charged with carboxy methyl groups, sulfonate groups, sulfate groups or combination thereof. 7. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide is levorotary. 8. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide has a molecular weight from 5,000 to 1,000,000. 9. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide has a molecular weight from above 25,000. ^10. The sulfated polysaccharide as claimed in claim 9, wherein the sulfated polysaccharide has a molecular weight from above 40,000. 11. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide has a molecular weight greater than 500,000 for topical application. 12. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide comprises D- glucopyranose residues linked by a-1, 6 linkages. 13. The sulfated polysaccharide as claimed in claim I, wherein the sulfated polysaccharide comprises L- glucopyranose residues. 14. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide is sulfated dextrin. 15. The sulfated polysaccharide as claimed in claim 1, wherein the sulfated polysaccharide is not dextrin sulfate, cyclodextrin or carrageenam.

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