The present invention relates to a process for the cure and control of Diabetes mellitus using natural products isolated from Perna viridis.

FIELD OF THE INVENTION

The present invention relates to a process for the cure and control of Diabetes mellitus using natural products isolated from Perna viridis. More particularly, the present invention relates to a process for the cure and control of Diabetes mellitus using natural products from Perna viridis.

BACKGROUND OF THE INVENTION

The birth and death of Pancreatic .beta.-cells is of obvious importance to the cure of Diabetes mellitus. For many years, .beta.-cells were considered terminally differentiated and unchanging after birth. However, based on the experimental animal models, the concept that .beta.-cell mass is not static but dynamic with ability to compensate for maintenance of glucose homeostasis has been proposed. In this context, search for the factors that lead to direct differentiation of .beta.-cells from precursor .beta.-cell mass in the diabetic individual is important to find out novel approaches to control and cure of diabetes by natural products obtained from the Indian marine bivalves Perna viridis.

The tissue extract from cellophan wrapped pancreas has been shown to possess trophic activity on pancreatic tissue inducing islet cell regeneration.sup.1. The active compound of cellophane wrapped pancreatic extract which is called as `ilotropin` has also been shown to be effective in reversal of Streptozotocin induced diabetes by induction of islet neognesis.sup.2. The regenerating pancreas is being shown to posses islet neogenetic activity leading to reversal of experimental diabetes.sup.3

The process of preparation of extract was developed for the first time by the Russian scientists. A patent was also filed on a process developed by Russian scientists vide Patent No. RU 2043109. The extract prepared from the Indian green mussel (Perna viridis) has previously been found to be active against all influenza, herpes and hepatitis viral strains. The extract is also found to possess not only prophylactic efficacy for protection from several viral diseases but it also shows a high therapeutic activity against these diseases. The process of preparation of extract was developed by NIO, Goa, in collaboration with the Russian Scientists.

The tissue extract from cellophane wrapped pancreas has been shown to possess trophic activity on pancreatic tissue inducing islet cell regeneration (Rosenberg L. and Vinik, A. I. 1989, Induction of endocrine cell differentiation: A new approach to management of diabetes, J. Lab. Clin. Med., 114:75-83). The active compound of cellophane wrapped pancreatic extract called as `ilotropin` has also been shown to be effective in reversal of streptozotocin induced diabetes by induction of islet neognesis (Rosenberg, L. Duguid, W. P. Healy, M. Clas, D. and Vinik, A. I. 1992, Reversal of diabetes by the induction of islet cell neogenesis, Transplant Proc., 24: 1027-1028). The regenerating pancreas is being shown to possess islet neognesis activity leading to reversal of experimental diabetes (Hardikar, A. A. and Bhonde, R. R., 1999, Modulating experimental diabetes by treatment with cytosolic extract from the regenerating pancreas, Diabet. Res. Clin. Pract, 46: 203-211).

OBJECT OF THE INVENTION

Accordingly it is an object of the invention to eliminate the need to use pancreatic preparations from animals for the reversal of diabetes.

It is another object of the invention, to assess the islet protecting activity of the mussel hydrolysate.

It is another object of the invention to determine the hypoglycemic and islet neogensis activity of the hydrolysate.

SUMMARY OF THE INVENTION

The invention provides a process for the cure and control of Diabetes mellitus using natural products from Perna viridis.

The present invention relates to the use of an extract of Indian green mussel for cure of diabetes mellitus due to the hypoglycemic activity and pancreatic regeneration potential of the extract prepared from the green mussel, said use comprising diluting extract by 50 times with 1.times.PBS, adjusting the pH to 7.2, adding extract to mouse islet cultures, injecting the extract to STZ diabetic mice and observing the blood glucose level of treated and untreated mice to ensure hypoglycemic activity.

The invention also relates to the use of natural products from Perna viridis for the cure and control of Diabetes mellitus.

In one embodiment of the invention, the extract permits elimination of pancreatic preparation from the animals, examination of the role of putative factors from the extract in preventing .beta.-cell apoptosis and evaluation of the mechanisms of action of the extract probably in curing diabetes.

In another embodiment of the present invention, hypoglycemic activity and pancreatic regeneration potential of the extract prepared from the green mussel is done with diluting the extract with 1.times.PBS by 50 times.

In another embodiment of the invention, the pH of the extract is adjusted to 7.2 with alkali (1 N NaOH)

In a further embodiment of the invention, the islets isolated from the mouse pancreas are cultured in six well plate in presence of RPMI-1640 supplemented with 10% diluted extract.

In a further embodiment of the invention, six well plates are incubated in a Co.sub.2 incubator at 37.degree. C. for 15 to 30 days to determine the viability integrity and attachment of the islets to the plates.

In another embodiment of the invention, the mussel hydrolysate has high protein content and contains no glucose.

In another embodiment of the invention, the mussel hydrolysate supports attachment and proliferation of AR42J cell line (pancreatic adenocarcinoma cell line) in a very low concentration (dilution 1:800 and 1:400) and in absence of FCS.

In a further embodiment of the invention, the mussel hydrolysate maintains the viability of pancreatic acinar cells.

In a further embodiment of the invention, 0.2 ml of extract is injected with the help of a sterilized disposable syringe to STZ induced diabetic mice.

In a further embodiment of the invention, the blood glucose level of treated and untreated mice is estimated using a glucometer to check hypoglycemic activity.

DETAILED DESCRIPTION OF THE INVENTION

According to the present invention, a process for the cure and control of Diabetes mellitus using natural products from Perna viridis has been found to be non-toxic and cyto-protective to isolated Islets of Langerhans from mouse pancreas. The extract is also found to posses islet neogenesis activity as evidenced by the reversal of the experimental diabetes in mice.

Preparation of Extract Marine Bivalves:

Live green mussels collected from the natural environment were deshelled and meat and mantle fluid was removed. The mixture of mantle fluid and meat was fermented at 40.degree. C. for two hours, the distillation and digestion process was carried out with concentration hydrochloric acid at 100+5.degree. C. for 20 hours. The resultant solution was neutralized with an alkali at room temperature to achieve a pH of 5.6 The active extract was isolated by keeping the solution in a separating flask and carefully removing the middle part of the solution after allowing the extract for settlement for 15 days.

Methodology

Hypoglycemic activity and pancreatic regeneration potential of the extract prepared from the green mussel was done by diluting the extract with 1.times.PBS by 50 times. The pH of the extract is adjusted to 7.2 with alkali (1 N NaOH).

The islets isolated from the mouse pancreas are cultured in six well plate in presence of RPMI-1640 supplemented with 10% diluted extract. The well plates are incubated in a CO.sub.2 incubator 37.degree. C. for 15 to 30 days to see the viability integrity and attachment of the islets to the plates. The 0.2 ml of extract is injected with the help of a sterilized disposable syringe to STZ diabetic mice.
 

Claim 1 of 21 Claims

1. A method for treatment of a patient with Diabetes mellitus comprising administering a pharmaceutically effective amount of a mussel extract to the patient, said mussel extract being a hydrolysate of meat and mantle fluid from Perna viridis.
 

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