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GOLD NANO PARTICLES FOR USE IN THERAPEUTIC METHODS

FIELD OF INVENTION

The invention relates to a method in targeted cell (we didn’t target cells) or tissue

5 destruction by thermal energy.

BACKGROUND OF INVENTION

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Start with obesity incidence around the world. How come people get fatter now? The problems

10 and what it can lead to. Do people generally care about this? Concern about health or look?

What’s the drawback on the nation? How much money has people spent on weight loss?

Goreng a bit la brother, im sure you can. End your paragraph stating that obesity – looking fat is

an important issue around the world. And wouldn’t it be nice if we can have an easy, safe & not

so invasive technology to get rid of the belly? People would spend money for this technology.

15 But so far, no technology has achieved these.

Advancements in the science of Nanotechnology have given us a host of new materials

and a deeper understanding of their properties. Among these novel materials are gold

nanoparticles. An interesting property of these nanoparticles is their ability to convert photonic

20 energy into thermal energy (ref). This is achieved through the phenomenon of plasmonics,

which is the study of oscillation of free electrons.

Every nanoparticle is surrounded by a cloud of free electrons. When a photon interacts

with this cloud of electrons they oscillate. This oscillation of the cloud of electrons generates

25 heat that is dissipated to the surrounding area. Lasers are a form of photonic radiation.

Therefore, when a solution of gold nanoparticles is exposed to a source of photonic radiation

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(i.e. a laser) the solution will increase in temperature (ref). Expand – describe a study in which

the temp increases – to what C – can it reaches >5C?. The cells (all cells?) in the human body

can only survive for a few minutes at temperatures 50 Celsius above our normal core

temperature of 370 Celsius, this heightened temperature cause’s damage to the cell membrane

5 leading to cell membrane lysis. This directly leads to cell death as a cell cannot survive without

a membrane.

This phenomena has up till now been applied in the treatment of cell proliferation

disorders, mainly cancer. The hope is that by treating the nanoparticles with suitable antibodies

10 that are specific to the type of targeted cell, the “targeted” destruction of the cancer cells can be

achieved. Expand – what type of cancer, in vitro/in vivo, what did the study show?, was it

efficient?, etc) .This method requires that the nanoparticle bind specifically to the targeted tissue

in order to cause tissue destruction. Research has found that the mere presence of

nanoparticles and a laser source are not enough to achieve cell destruction (Lowery, A.R.;

15 2006; Immunonanoshells for targeted photothermal ablation of tumor cells; International Journal

of Nanomedicine 2006:1(2) (Quick question, by reading this, one will not be able to come up or

have the slightest idea to come up with your invention right? If so, we will maintain this but if this

is obvious then it will be advisable not to include this.). Do you mean that people can copywhat Formatted: Font: Not Italic

we do from this info? I don’t think so. I suggest we expand a bit more on the study, but leave the

20 details on the methodology,

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If I were you I would state my hypothesis here to link between the paragraph above and below.

Seems that there is no connection. The hypothesis would be something like this – based on the

study by Lowerry we hypothesized that the photothermal ablation could also be applied to fat

25 cells. We should only state this if it doesn’t risk our patent application. Formatted: Font: Not Italic
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The fat cell is the body’s energy store. Excess calories consumed is converted to fat and

stored inside fat cells. Although the number of fat cells in the body is fixed, the size of one fat

cell is only limited by how much fat is stored within it. The problem of excess adipose tissue is a

concern for a large size of the worlds’ population. Diet and exercise should be the main tools

5 used to deal with this problem. Having said that, it is acknowledged that there are some with

“stubborn fat”, areas on the body that don’t respond well to diet and exercise.

Lipolysis, which refers to the breakdown of fat cells has been used as a cosmetic

procedure to rid the body of unwanted adipose tissue deposits for decades. The theory behind

10 the treatment states that if an agent could be introduced to the adipose tissue to cause

dissolution of fat cell membranes then this will lead to death of the fat cell itself (ref). These

dead fat cells will be removed by the body naturally, as is any dead tissue in the normal cell

turnover cycle. This process liberates the fat stored within the fat cells to be used up as fuel by

the body. The end result is that the unwanted fat/adipose tissue is gotten rid of and more

15 pleasing aesthetic look is achieved.

The current standard for removing unwanted fat tissue is liposuction. It is a surgical

procedure and like any surgical procedure comes with its set of risks and complications.

Describe how liposuction is being done. Focus on the gory part of the procedure. Liposuction is

20 suitable when larger amounts of fat are desired to be removed. Highlight the death incidence

happened in Maaysia and around the world – need ref for this.
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For targeted removal of small amounts of fat of less than 100 ml, injection lipolysis is a

popular method. In this method, a chemical phosphotidylcholine is injected along with a solvent

(due to the fact that phosphotidylcholine is not water soluble) to the desired fat tissue area. This

causes breakdown of fat cell membranes and leads to fat cell death. Although this method is

5 widely practiced, it is classified under “off label use” of the drug Lipostabil (phosphatidylcholine).

Lipostabil is only approved by the FDA for the treatment of liver cirrhosis (ref). The addition of a

solvent, which by itself can cause lipolysis, to the agent leads to a confusion of the mode of

action of how lipolysis is achieved. Since the agent used is active as soon as it comes in contact

with any living tissue, the risk of damaging healthy or untargeted tissue is ever present due to

10 possible technical incompetence on the part of the one administering the treatment (ref). Any

example to support this?

With this, injection lipolysis will remain as a treatment that cannot be advertised and

promoted under FDA regulations for lack of clear scientific basis for its outcomes. Therefore,

15 there is a need for a new approach of targeted cell destruction, in particular, fat cells which

poses less risk but also sustainable for industries which require them. Awesome.

SUMMARY OF INVENTION

20 The invention provides a method for cell or tissue destruction using thermal energy, in

particular fat cells, for therapeutic or cosmetic purposes. The invention provides method for

inducing cell lysis wherein the steps includes (a) administering gold nanoparticle to the fat?

cells; (b) exposing the gold nanoparticles to a radiation source at a particular wave length?; and

(c) destroying the cells using the radiation exposed gold nanoparticle, wherein the gold

25 nanoparticle does not attach to the cells and emits heat for cell destruction when exposed to

radiation. The gold nanoparticles emit heat due to electron oscillation and the electron
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oscillation is produced by interacting electrons surrounding the gold nanoparticles with photons

from the radiation. The radiation source is a photonic radiation which includes near-infrared. The

method is not only applicable to fat cells in particular but also cells or tissues such as connective

tissue, nerve cells and blood vessels. (People may concern that this method can also kill non

5 intended cell – try to defend this)

BRIEF DESCRITION OF THE FIGURES

FIG. 1 refers to the effect on the adipocytes using gold nanorods (GNR), viewed after treatment

10 with calcein. (Can we have a little explanation on what is the effect?) Yes we should. The Formatted: Font: Not Italic
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readers will also want to know what is the calcein for. Magnification? What microscope?
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FIG. 2 refers to the effect on the adipocytes using near infra-red (NIR) laser, viewed after

treatment with calcein. (Can we have a little explanation on what is the effect?) Same as above. Formatted: Font: Not Italic

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FIG. 3 refers to the effect on the adipocytes using both gold nanorods (GNR) and near infra-red

(NIR) laser, viewed after treatment with calcein. (Can we have a little explanation on what is the

effect?) Same Formatted: Font: Not Italic

20 FIG. 4 refers to the effect on the adipocytes using both gold nanorods (GNR) and near infra-red

(NIR) laser, viewed after treatment with propidium iodide. (Can we have a little explanation on

what is the effect?) same – explain why use PI.

No where describing in detail the procedure we use. Are we not advised to do so? Formatted: Font: Not Italic

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DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention provides a method for cell or tissue destruction using thermal

energy, in particular fat cells, for therapeutic or cosmetic purposes. Hereinafter, this

specification will describe the present invention according to the preferred embodiments of the

5 present invention. However, the following example is to be understood that limiting the

description to the preferred embodiments of the invention is merely to facilitate discussion of the

present invention and it is envisioned that those skilled in the art may devise various

modifications and equivalents without departing from the scope of the appended claims.

10 The invention provides a method which uses the combination of gold nano-rods (GNR)

and heat from near infra-red (NIR) or any source of radiation to result in cell destruction, in

particular fat cells. This invention achieves cell destruction without the need for binding of

nanoparticles to the targeted cell type. The main cell type targeted is, but not limited, adipocytes

or fat cells.

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The basis of the lipolytic ability of GNRs and NIR lasers is due to plasmonics.

Plasmonics is a phenomenon that occurs when the photonic energy of the NIR laser interacts

with the cloud of electrons surrounding a GNR. This interaction causes the cloud of electrons

surrounding the GNRs to oscillate which in turn generates energy. This energy is then

20 dissipated as heat to the surrounding, which leads to an increase in temperature in the area

immediate to the GNRs. The energy which dissipated as heat that causes lipolysis. Cells in our

body can only survive a temperature increase of 5° C for 10 minutes duration before the cell

membrane breaks down and leads to cell death. In our study we never did prove that the effect

was not due without the binding of GNR to fat cells- in which you claimed earlier. Does this

25 paragraph somehow prove that there is actually no binding based on the heat property emitted
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from the GNR? If it does, please state so. How close the GNR needs to be near the cells for the

heat to cause the effect?

The method is carried out with a determined and preferred concentration of GNRs and

5 exposure time to NIR laser and with this, the use of these two elements can lead to a new

method of lipolysis. This method can be used in various applications such as therapeutic

purposes and in particular for cosmetic purposes.

As mentioned, the principle in this cell destruction is the combination of both the GNRs

10 and NIR. The application of GNRs and NIR individually does not result in cell destruction (Fig 1

&2?). The administered GNRs will be present in the layer of fat cells however; adsorption of

GNRs into the fat cells is not required (how can we support this?). Gold nanoparticles are a

class of nano materials that are able to convert photonic energy into thermal energy. This

occurs when the photons from the NIR interacts with cloud of electrons present around the gold

15 nanoparticles. This causes oscillation which is eventually converted into thermal energy. The

thermal energy or heat which is emitted causes the fat cells to break down and destruct due to

increase of temperature surrounding the cells.

Experimental data as to the efficacy of this invention was obtained by an in vitro

20 demonstration using cultured human adipocytes, a laser emitting in the near infra red frequency

and an appropriate concentration of gold nanorods solution. The choice of the near infra red

frequency spectrum for the laser was influenced by the ability of the laser light at this spectrum

to penetrate through human tissue without causing any damage. The human fat cells were

cultured from a pre-adipocyte to mature adipocyte stage in a sterile environment. The cultured

25 fat cells were then divided into three groups. One group was treated by administering only gold
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nanorods; another was treated by only exposing them to the near infra red spectrum laser while

the third group received both the gold nanorods and near infra red spectrum laser.

It is pertinent to note that the laser treatment of the last group (gold nanorods and laser)

5 commenced immediately after the gold nanorods solution had been administered. This ensures

that the method of action of this invention does not require attachment of gold nanoparticles to

the target cell as review of the literature shows (which literatures, show references) that a

minimum of 2 hours is required for attachment of nanoparticles to cell membranes. Fluorescent

dye cell viability assay was used to determine adipocyte destruction.

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The results clearly demonstrate that the destruction of the adipocytes only occur in the

presence of both gold nanorods and laser irradiation. (please refer to attached pictures).

For example, a patient may remove fat deposits in targeted areas of her body by

15 undergoing this method as presented in the invention. Based on the amount of fat that is

desired to be removed, the clinician then administers the appropriate amount of GNRs solution

(in millilitres) to the targeted area. The GNR solution is administered through an injection into

the subcutaneous fat layer and taking care not to go any deeper in the layer where the targeted

cells are. Once the GNRs are administered to the area and without attachment to the targeted

20 cells, the GNR is then exposed almost immediately to a NIR laser of the appropriate frequency

for duration of 10 minutes. An added benefit of this method over existing methods is the safety

aspect. As mentioned, the Lipostabil and the solvent used can lead to unwanted damages such

as muscle damage when administered incorrectly.

25 The use of GNRs prevents the problem of probable unwanted damages as the GNRs do

not have an effect on any tissues when being administered to the cells. The GNRs are only
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activated to emit heat when there is the presence of photonic energy from a NIR laser. The

depth of the laser penetration in turn is determined by its frequency. Hence, if the GNRs were to

be administered by mistake into the muscular layer or any layer of tissues, it would not result in

tissue damage as the NIR lasers’ energy will not be able to penetrate the said layer. Good stuff.

Therefore, this method presents a promising alternative to the current technology in cell

or tissue, in particular fat cells, destruction using thermal energy for therapeutic or cosmetic

purposes.
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CLAIMS

1. A method for inducing cell lysis wherein the steps includes:

a. administering gold nanoparticle to the cells;

5 b. exposing the gold nanoparticle to a radiation source; and

c. destroying the cells using the radiation exposed gold nanoparticle

wherein the gold nanoparticle does not attach to the cells and the gold nanoparticle emits

heat for cell destruction when exposed to radiation.

10 2. The method as claimed in claim 1 wherein the cells are from one or more from the group of

fat cells, connective tissue, nerve cells and blood vessels.

3. The method as claimed in claim 1 wherein the radiation is a photonic radiation which

15 includes near-infrared.

4. A method for inducing fat cell lysis wherein the steps includes:

a. administering gold nanoparticle to the cells;

b. exposing the gold nanoparticle to a radiation source; and

20 c. destroying the cells using the radiation exposed gold nanoparticle

wherein the gold nanoparticle does not attach to the cells and the gold nanoparticle emits

heat for cell destruction when exposed to radiation.

25 5. The method as claimed in claim 1 wherein the radiation is a photonic radiation which

includes near-infrared. Formatted: Font: Bold

5. Claim 4 & 5 – similar to 1 & 3? Formatted: Indent: Left: 0.25", No bullets or

numbering
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ABSTRACT

GOLD NANO PARTICLES FOR USE IN THERAPEUTIC METHODS (I thought we Formatted: Font: Not Bold

gonna only sell this idea for fat cells? Shdnt we focus?)

5 The invention provides a method for cell or tissue destruction using thermal energy, in

particular fat cells, for therapeutic or cosmetic purposes. The invention provides method for

inducing cell lysis wherein the steps includes (a) administering gold nanoparticle to the cells; (b)

exposing the gold nanoparticles to a radiation source; and (c) destroying the cells using the

radiation exposed gold nanoparticle, wherein the gold nanoparticle does not attach to the cells

10 and emits heat for cell destruction when exposed to radiation. The gold nanoparticles emit heat

due to electron oscillation and the electron oscillation is produced by interacting electrons

surrounding the gold nanoparticles with photons from the radiation. The radiation source is a

photonic radiation which includes near-infrared. The method is not only applicable to fat cells in

particular but also cells or tissues such as connective tissue, nerve cells and blood vessels.

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FIG. 1, 2, 3 and 4.