# US 8,147,846 B2 · Method for the treatment of airway diseases

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- Number: US 8,147,846 B2
- Document ID: US8147846B2
- Status: Granted
- Kind: B2
- Application number: 11/296,243
- Filed: 2005-12-07
- Published: 2007-06-07
- Granted: 2012-04-03
- Earliest claimed: 2005-12-07
- Assignees: Board of Trustees of Michigan State University
- Inventors: Alberto Leonel Mendoza; Robert Louis Glass
- Web page: https://patentgrep.com/patent/US8147846B2

## CPC

- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

A method for the treatment of allergic airway diseases in mammals related to Pythiosis insidiosum is described. The method relies upon an immunotherapeutic product of pythium proteins as antigens. The method is particularly useful in equine cicatrix.

## Claims

### Claim 1

A method for treatment of equine allergic airway disease, which comprises:
injecting or orally dosing the equine with a treatment effective amount of an immunotherapeutic product comprising at least one isolated protein of Pythium insidiosum which treats the disease;
said injecting or oral dosing occurring in multiple doses over a time period sufficient to lessen breathing difficulties.

### Claim 2 (depends on 1)

The method of claim 1 wherein the treatment time period sufficient is at least 21 days.

### Claim 3 (depends on 1)

The method of claim 1 wherein the immunotherapeutic product comprises in a sterile aqueous solution an admixture of:
(a) intracellular proteins separated from disrupted cells of Pythium insidiosum grown in culture medium and from extracellular proteins; and
(b) extracellular proteins comprising 28, 30 and 32 kD proteins as determined by SDS-PAGE electrophoresis in a supernatant from the culture medium for growing the cells of the Pythium insidiosum which has been separated from the cells, wherein the proteins in the admixture of the separated intracellular and extracellular proteins have been mixed with sterile distilled water and dialyzed.

### Claim 4 (depends on 3)

The method of claim 3 wherein the extracellular proteins have been provided by growing cells of the Pythium insidiosum in a culture medium, killing the cells by chemical or biological agent, and separating the killed cells from the culture medium containing the extracellular proteins and the intracellular proteins which have been provided by disrupting the killed cells and removing the disrupted cells from the intracellular proteins.

### Claim 5 (depends on 4)

The method of claim 4 wherein the killed cells have been disrupted by sonication or cryogenic grinding.

### Claim 6 (depends on 1)

The method of claim 1 wherein the Pythium insidiosum is deposited as ATCC 74446.

### Claim 7 (depends on 3, 4)

The method of any one of claim 3 or 4 wherein the culture medium used for growing the cells is Sabouraud dextrose broth.

### Claim 8 (depends on 4)

The method of claim 4 wherein the killed cells of Pythium insidiosum have been killed with thimersol, phenol or other chemical or biological agent after growing in the culture medium.

### Claim 9 (depends on 4)

The method of claim 4 wherein the intracellular proteins are separated from the disrupted cells by centrifugation or filtration.

### Claim 10 (depends on 4)

The method of claim 4 wherein the proteins which are admixed have been separated from the supernatant by being precipitated together using acetone and then the precipitate is dispersed in sterile distilled water.