A technology to simultaneously produce broad-spectrum antiviral polysaccharides from marine microalgae, as well as the omega-3 fatty acid, DHA.

Summary:
The invention provides broad-spectrum antiviral extracellular polysaccharides from the marine, heterotrophic, microalgae called thraustochytrids. These microalgae also produce the omega-3 fatty acid, DHA. Therefore, this is an economical process to produce two valuable compounds from safe microorganisms.

Full Description:
Any of the several candidate strains available with Myko Tech is grown in a suitable culture medium in fermentors. After 4 to 7 days of growth, the biomass is harvested for extraction of oils containing the omega-3 polyunsaturated fatty acid (PUFA), docosahexaenoic acid (DHA). The culture filtrate is used to extract extracellular polysaccharides.
The importance of omega-3 fatty acids, particularly DHA for cardiovascular health is now very well recognized. Although fish oils are presently the largest source for these oils, microbial sources from thraustochytrid micro-algae are gaining a large market because of their more favourable oil characteristics. Myko Tech’s technology helps in production of omega-3 rich oils as one of the two products, simultaneously, the other being antiviral polysaccharides.
The five important viruses addressed by Myko Tech’s technology are the following.
(1) Enterovirus: A species of Enterovirus causes polio for which a vaccine exists. Infections by other species can result in a wide variety of symptoms ranging from mild respiratory illness (common cold), hand, foot and mouth disease, acute hemarrhogic conjunctivitis, asptic meningitis, mycorditis, severe neonatal sepsis-like disease and acute flaccid paralysis. There are presently no drugs for enteroviruses.
(2) Retrovirus. There is no effective vaccine against HIV infection and the disease is one of the most serious medical problems in view of its rapidly expanding HIV epidemic. Although there is an impressive development of anti-HIV drugs, new patterns of resistance and need for fewer side effects will require new drugs.
(3) Cytomegalovirus: CMV infection is a threat to AIDS or transplant patients because of their immunodeficiency. CMV causes a variety of conditions from retinitis to pneumonitis. Although HIV patients undergoing a combination treatment are less prone to CMV diseases, those undergoing organ transplant still face the risk of CMV infections. Ganciclovir and foscarnet have been used for treatment of CMV infections and recent studies show valaciclovir to have a prophylactic effect against CMV disease in transplant. Development of resistance to aciclovir and ganciclovir in CMV-infected patients will require the need to develop new drugs. Foscarnet therapy is associated with side effects and problems of administering.
(4) Adenovirus. ‘Common cold’ infections are mostly caused by adenovirus. Adenovirus infections could be life-threatening for immunodeficient patients.
Since there is no efficient vaccine for adenoviruses, development of a drug remains important.
(5) Herpes simplex virus (HSV). Labial herpes (cold sores) and genital herpes are recurrent diseases afflicting a large number of individuals worldwide. HSV-1 and 2 can also cause eye disease, encephalitis and generalized infections in immunodeficient patients and newborns. There is no vaccination against these. Acyclovi, Valaciclovir, Penciclovir, Vectavir, Famciclovir, Famvir and Foscarnet are drugs to treat herpes. Both acyclovir, and its more bioavailable prodrug valaciclovir, have been shown to be effective in prophylaxis against episodes of genital herpes, thereby preventing painful episodes of genital sores. Penciclovir, and its prodrug famciclovir, show benefit in prophylaxis against genital herpes. However, the therapeutic effect of topical aciclovir and penciclovir against recurring episodes of labial cold sores and recurring genital herpes is limited to about a 10% reduction in healing time, even if treatment is initiated during prodromal symptoms. New drugs to treat HSV will be very useful.

An additional virus for which a similar polysaccharide has been found to be effective is the hepatitis viruses.
Present sources of antiviral polysaccharides from marine sources and drawbacks

• Marine cell wall polysaccharides from various micro- and macroalgae have been shown to be selective inhibitors of various enveloped viruses, including herpes simplex virus, cytomegalovirus, vesicular stomatitis virus, and human immunodeficiency virus and Herpes Simplex Virus (HSV) (Baba et al., 1988). Others have shown antiviral activities, such as against Herpes Simplex Virus (HSV) (Huleihel et al., 2001, 2002; Talyshinsky et al., 2002; US Patent 5089481; Harden et al., 2009).
• However, extraction from cell walls is a cumbersome process and polysaccharides from large, macroalgae is an ecologically unsustainable process.
• Extracellular polysaccharides or EPS that are secreted outside cells are easier to extract and use. EPS can be extracted in a pure form more easily. Antiviral EPS using microorganisms grown in large volumes in fermenters and which secrete EPS has several advantages as the process is cost-effective, economical and production can be carried out at large scale, while being environmentally sustainable as well. Various bacteria and some marine algae produce antiviral, extracellular polysaccharides, active against various viruses, including HSV and Influenza A viruses (Geresh and Arad, 1991; Okutani, 1992; Matsuda et al., 1999; Arena et al., 2006).
These studies show the enormous potential of using EPS to treat viral diseases. The production of such EPS from a microorganism, if combined with the use of the cell biomass for yet another useful purpose will be extremely beneficial.

Literature:

Baba M, Snoeck R, Pauwels R, de Clercq E. 1988. Sulfated polysaccharides are potent and selective inhibitors of various enveloped viruses, including herpes simplex virus, cytomegalovirus, vesicular stomatitis virus, and human immunodeficiency virus. Antimicrob Agents Chemother 32, 1742-2745;
Patent applications WO 2009027057 and WO 20090305.
Huleihel M, Ishanu V, Tal J, Arad S: Antiviral effect of microalgal polysaccharides on Herpes simplex and Varicella zoster viruses. J. Appl. Phycol. 2001, 13:127-134),
Huleihel M, Ishanu V, Tal J, Arad SM. 2002. Activity of Porphyridium sp. polysaccharide against herpes simplex viruses in vitro and in vivo. J Biochem Biophys Methods. 2002 50:189-200)
US Patent 5089481;
Harden EA, Falshaw R, Carnachan SM, Kern ER, Prichard MN, 2009. Virucidal activity of polysaccharide extracts from four algal species against herpes simplex virus. Antiviral Research 83: 282-289).
United States Patent 5089481 of 1992
Talyshinsky MM, YY Souprun and MM Huleihel. 2002. Antiviral activity of red micro-algal polysaccharides against retroviruses. Cancer Cell International 2: 1-7). However, extraction from the cell wall of the algae is a cumbersome process.
Okutani K. 1992. Antiviral activities of sulfated derivates of a fucosamine containing polysaccharide of marine bacterial origin. Nippon Suisan Gakkaishi 58:927–30;
S. Geresh and S. Arad, 1991. The Extracellular Polysaccharides of the Red Microalgae: Chemistry and Rheology Bio resource Technology 38: 195-201;
Arena A, Maugeri TL, Pavone B, Iannello D, Gugliandolo C and Bisignano G. 2006. Antiviral and immunoregulatory effect of a novel exopolysaccharide from a marine thermotolerant Bacillus licheniformis. International Immunopharmacology 6: 8– 13).
Matsuda M, Shigeta S, Okutani K. 1999. Antiviral Activities of Marine Pseudomonas Polysaccharides and Their Oversulfated Derivatives. Marine Biotechnology 1: 68–73).

Myko Tech’s process

1. Myko Tech uses strains of the marine, heterotrophic, single-celled microalgae called thraustochytrids which typically produce high levels of the omega-3 PUFA, DHA, as well as high amounts of extracellular polysaccharides.
2. Cultures are grown in simple, inexpensive culture media with an organic carbon source, organic or inorganic nitrogen source, vitamin and trace elements. The media are made up in diluted seawater.
3. Optimal culture and fermentation parameters for high yields are used.
4. After growth for 5 to 6 days, cell biomass is separated from the culture filtrate. The biomass may be used for extraction of lipids rich in DHA.
5. The culture filtrate may be concentrated and the EPS precipitated. The EPS may be purified by column chromatography. This has the potential to be used as a topical antiviral agent.
6. Purified EPS may be fractionated further to obtain the active antiviral fraction. This has the potential to be used as an antiviral drug.
7. The EPS has been tested against the model viruses enterovirus 71 in Vero cell lines, Human Adenovirus 5 in A549 cell line, Retrovirus XMRV in PG54 cell line and Human Cytomegalovirus AD-169 in MRC5 cell lines, as well as against Herpes Simplex Virus-1 in Vero cell lines.

Advantages of Myko Tech’s process

• Myko Tech’s process is based upon the heterotrophic group of marine single-celled protists, the thraustochytrids, belonging to the Labyrinthulomycetes that produce sulphated EPS, as well as the omega-3 polyunsaturated fatty acid (PUFA), docosahexaenoic acid (DHA) (Raghukumar, S. 2009. Thraustochytrid marine protists: production of PUFAs and other emerging technologies. Marine Biotechnology, 10: 631-640. Therefore, two products are possible simultaneously.
• Myko Tech possesses several strains that grow rapidly, easily produce up to 100 g of dry weight biomass per liter medium and 10 g of EPS per liter medium, making this process of producing two useful products at the same time highly feasible and economical.
• Since thraustochytrid are considered safe for human use, the EPS are likely to be nontoxic for use as antiviral drugs.
• Myko Tech has filed a PCT application for antiviral EPS against enteroviruses, adenoviruses, retroviruses and cytomegalovirus. An Indian Patent has been filed for treating herpes virus.
• US Patent 8,232,090 has earlier described the production of extracellular polysaccharides showing anti-hepatitis virus activity in a strain of Schizochytrium limacinum MTCC 5249.
Indian Patent Application 3257/MUM/2010 provides a process to produce an anti-herpes polysaccharide by the thraustochytrid strain of Schizochytrium limacinum MTCC 5249.
Myko Tech’s Publications

1. 2009. Raghukumar S. Thraustochytrid marine protists: production of PUFAs and other emerging technologies. Marine Biotechnology, 10: 631-640.

Problem this idea/invention addresses:
The invention provides potential antiviral drugs in the form of extracellular polysaccharides from a group of marine, micro-algae that also produce another valuable product, namely docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid. The invention thus provides two products simultaneously, making it highly economical.
This original technology is based on the international expertise of Dr Seshagiri Raghukumar on the marine heterotrophic, microalgae, the thraustochytrids, which also produce omega-3 polyunsaturated fatty acids. The extracellular polysaccharides (EPS) have advantages over many existing drugs because of their natural, apparently non-toxic nature from safe organisms, their broad-spectrum activity and low cost of production. A further advantage is that while the extracellular polysaccharides can be harvested from the culture filtrate, a second valuable product, namely docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid can be extracted from the biomass. DHA is an important nutraceutical for preventing cardiovascular development. The technology covers the use of these EPS against enteroviruses, adenoviruses, cytomegaloviruses, retroviruses and herpes simplex virus.

Auxiliary products or services for sale:
Offer type:

Negotiable.
• Joint collaboration to complete development of the products and take it to the market.
• Venture capital to Myko Tech to complete development.
• Licensing or sale of technology at the present stage.


Technology for sale or license: Patented microbial strain, technical knowhow, patent rights.

Cost: Negotiable. Based on which of the above offers is accepted.

Company
Myko Tech Private Limited,
313, Vainguinnim Valley,
Dona Paula, Goa – 403 004.

Contact person: Dr Seshagiri Raghukumar
E-mail: [email protected]
Phone Number: +91(0832)256070
Mobile: +919890450007

Asking price: [CONTACT SELLER]
Available for consultation? Yes

Invention #11880
Date posted: 2013-09-29


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