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Immunokine composition and method

a technology of immunomodulatory compounds and compositions, applied in the direction of immunological disorders, peptide/protein ingredients, peptide sources, etc., can solve the problems of rapid development of viral strains, inability to infect cells, and ba-l macrophage tropic strain, so as to prevent the initial hiv infection of the individual, prevent the infection of the cell, and limit the spread of the virus

Inactive Publication Date: 2008-10-09
PHYLOMED CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0021]A composition of this invention is useful in preventing infection of a cell, both with in terms of treating existing HIV spread within an infected individual as well as preventing initial HIV infection of that individual. As such, the composition can be useful in limiting the spread of virus from one cell to another in an infected host and, if present, (i.e. circulating within a host) prior to exposure (but not productive infection) of a cell.

Problems solved by technology

However, the macrophage tropic strain Ba-L, is not capable of infecting cells which co-express both CXCR4 and CD4.
While these therapies, particularly when used in combination with one another, are effective, they are frequently short-lived in that viral strains rapidly develop that are resistant to one or more of the compounds used.

Method used

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  • Immunokine composition and method

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation of Gland Tissue for RNA Extraction

[0102]The following protocol was used to clone the gene encoding α-cobratoxin from the venom of Naja naja siamensis.

[0103](a) Recovery of Venom

[0104]Naja naja siamensis snakes were obtained from Siam Farms, Bangkok, Thailand. Animals were shipped to and housed at Ventoxin, Inc., Frederick, Md. USA. The venom glands from N. n. siamensis animals were surgically removed and used to prepare mRNA for generating a cDNA library. Snakes were placed on a schedule for milking (venom extraction). They were milked on day 1 and eight days later milked a second time. On the 2nd or 3rd day, they were anesthetized with sodium pentobarbital and their glands removed (Vandenplas et al., 1985). Gland tissue was quickly cut into small pieces and immediately frozen in liquid nitrogen. Samples were kept at −70° C. until use.

[0105](b) RNA Isolation

[0106]Total RNA was isolated from gland tissue by using a standard guanidinium / hot phenol method (Feramisco et al., ...

example 2

Recovery and Yield

[0139]A fermentation of a cytoplasmically-expressing clone harboring the gene encoding α-cobratoxin can be performed in a 5 L New Brunswick BioFlo III fermentor. The size of the fermentation can be scaled up or down depending on the requirement for product. For a 5 L batch, a frozen seed culture containing the alpha-cobratoxin construct is used to inoculate 10 ml of MGY media (see attached media recipe) in a test tube. After 18 to 20 hours growth at 30° C., 0.5 ml is used to inoculate 50 ml of MGY in a 250 ml flask. After 36 to 38 hours of growth, the entire 50 ml is used to inoculate the 5 L fermentor. The fermentation is performed in a basal salt medium with 26.7 ml 85% phosphoric acid, 0.93 g / L calcium sulfate-2H20, 18.2 g / L potassium sulfate, 14.9 g / L magnesium sulfate, 4.13 g / L potassium hydroxide, 40 g / L glycerol and 2 m / L of basal salts (PTM) are added. PTM basal salts consist of 2.0 g cupric sulfate, 0.08 g sodium iodide, 3.0 g magnesium sulfate, 0.2 g sodi...

example 3

Ozonation

[0143]Ozone (O3), a powerful oxidant, is used for water disinfection. In the course of the present invention, ozone treatment is preferably used to treat the recovered, inactive polypeptide in order to render it incapable of spontaneous reformation. Optionally, ozonated pure water can be used to itself selectively break the disulfide bonds of a formed polypeptide in order to provide an inactive, denatured, and stable form thereof.

[0144]Ozone treatment can be used to quickly provide microbial sterilization and disinfection, organic compound destruction, and conversion of iron or manganese salts to insoluble oxides which can be precipitated from the water. The major reaction byproducts are water, oxygen and carbon dioxide. For environmental and safety concerns, unreacted or residual ozone should be monitored. A number of UV spectrophotometric methods can be used to determine the level of ozone in water or physiological saline. Ozone has an absorption peak at 260 nm whereas ox...

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Abstract

A composition and method for preventing HIV infection of mammalian cells. One aspect of the invention relates to an anti-immunodeficiency virus immunokine capable of binding to a cellular protein in a manner that prevents HIV infection of that cell. The compositions can include either an active bioactive polypeptide, such as native cobratoxin, and / or an inactivated bioactive polypeptide, such as cobratoxin in which one or more of the native disulfide bridges have been prevented from forming. The term “immunokine” is used to refer to an inactivated bioactive polypeptide, whether inactivated by chemical, genetic, and / or synthetic means as described herein, with the proviso that a corresponding active bioactive polypeptides can be included where applicable (e.g., for in vitro use).

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a Continuation-in-Part of prior application Ser. No. 09 / 368,834, filed on Aug. 5, 1999, which is a continuation of prior application Ser. No. 08 / 908,212, filed on Aug. 7, 1997, now U.S. Pat. No. 5,989,857, which is a continuation of US patent application filed May 10, 1996 and assigned Ser. No. 08 / 644,399 for POLYPEPTIDE COMPOSITIONS AND METHODS, the entire disclosure of which is incorporated herein by reference.TECHNICAL FIELD[0002]The present invention relates to the treatment and prevention of viral infections, including HIV infections.BACKGROUND OF THE INVENTION[0003]The lack of an effective vaccine and the increase in antiretroviral drug treatment failures has led the HIV research community to continue the search for novel approaches to treat HIV infection. HIV can be inhibited at a number of different steps in its lifecycle within the cell or, alternatively, vaccines and immune based therapies can eliminate HIV-i...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00A61P37/00C07K1/113C07K14/46
CPCC07K1/1133C07K14/46A61P37/00
Inventor MUNDSCHENK, DAVID D.REID, PAUL F.
Owner PHYLOMED CORP
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