# US 8,465,939 B2 · Aging-related circulating particle-associated lipoprotein B oxidase (apoBNOX) and inhibitors thereof

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,465,939 B2
- Document ID: US8465939B2
- Status: Granted
- Kind: B2
- Application number: 13/039,170
- Filed: 2011-03-02
- Published: 2011-09-08
- Granted: 2013-06-18
- Earliest claimed: 2010-03-02
- Assignees: Nox Technologies, Inc.
- Applicants: D. James Morré
- Inventors: D. James Morré
- Web page: https://patentgrep.com/patent/US8465939B2

## CPC

- C12N 9/0036: {acting on NADH or NADPH (1.6)}
- A61K 31/05: Phenols {(cannabinoids A61K31/658)}
- A61K 36/87: Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- C12Q 1/26: involving oxidoreductase
- G01N 33/92: involving lipids, e.g. cholesterol {, lipoproteins, or their receptors (steroid hormones G01N33/743)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Provided is an age-related apolipoprotein B oxidase (apoBNOX) found tightly associated with the low-density lipoprotein particles and believed to be responsible for oxidizing lipoprotein particles and initiating atherogenesis. It causes damage by directly oxidizing the apolipoprotein B protein and indirectly oxidizing the lipids in the particles due to superoxide formation by the apoBNOX and its conversion into hydrogen peroxide. apoBNOX activity is inhibited by tyrosol and hydroxytyrosol and components of white wine, important components of French and Mediterranean diets, which seem to be a very good source of inhibitors of the apolipoprotein B oxidase. Agents comprising at least one naturally-occurring apoBNOX inhibitor and compositions lessen, ameliorate or treat disorders and complications resulting from cell damage caused by oxidation of apolipoprotein B. Also provided is an assay system comprising recombinant apoBNOX or an equivalent peptide and apolipoprotein B purified from human sera as a model to screen for agents and supplements that lower apoBNOX activity when administered orally.

## Claims

### Claim 1

A method of identifying an inhibitor of apolipoprotein B NADH oxidase (apoBNOX), said method comprising the steps of:
a) contacting a biological sample comprising apoBNOX and apolipoprotein B with a test compound in vitro to produce a contacted sample;
b) measuring apoBNOX activity in the contacted sample of step (a); and
c) identifying the test compound as an inhibitor of apoBNOX when the apoBNOX activity measured in step (b) is less than the apoBNOX activity of a comparison in vitro biological sample that has not been contacted with the test compound.

### Claim 2 (depends on 1)

The method of claim 1, wherein the apoBNOX activity is measured and identified as a reduction of ferricytochrome c.

### Claim 3 (depends on 1)

The method of claim 1, wherein the apoBNOX activity is measured by a reduction of NADH which is determined spectrophotometrically and identified as a decrease in absorbance at 340 nm.

### Claim 4 (depends on 1)

The method of claim 1, wherein the apolipoprotein B is provided in step (a) as low density lipoprotein particles.

### Claim 5 (depends on 1)

The method of claim 1, wherein the apolipoprotein B in step (a) is purified from low density lipoprotein particles.

### Claim 6 (depends on 1)

The method of claim 1, where the apolipoprotein B is provided in step a) as an equivalent peptide corresponding to the cellular binding region that normally forms a complex with apoBNOX and restricts access of the majority of arNOX inhibitors to the apoBNOX catalytic site.

### Claim 7 (depends on 6)

The method of claim 6, wherein the equivalent peptide comprises the amino acid sequence set forth in SEQ ID NO:3 or an amino acid sequence with at least 50%, 75%, 80%, 92%, or 96% amino acid identity thereto.

### Claim 8 (depends on 1)

The method of claim 1, wherein the apoBNOX is bound to the apolipoprotein B.