GRAPHENE BASED THERANOSTICS FOR TUMOR TARGETED DRUG/GENE DELIVERY AND IMAGING

Subhra Mohapatra

Patent

Atlas literature

Patent

US 9,675,714

GRAPHENE BASED THERANOSTICS FOR TUMOR TARGETED DRUG/GENE DELIVERY AND IMAGING

Subhra Mohapatra

Drawings

Patent drawings and their descriptions. Click a drawing to enlarge it.

Figure 1

Figure 1 shows one embodiment of synthesis of a doxorubicin-Chitosan functionalized magnetic graphene (CMG)-DNA nanoparticle

Figure 2

Figure 2 shows Fou ri ner Transform Infrared (FTIR) spectra of (a) chemically reduced 25 graphene oxide (C RGO), (b) CRGO- COO H, and (c) CMG 3 T| H Docket Number 292103-

Figure 3

Figure 3 shows magnetic properties of C MG with diﬀerent concentrations of Fe and a T₂ relaxometry map de ri ved from multi-TE-T₂ measurements

Figure 4

Figure 4 shows the viability of P C 3 cells treated with diﬀerent concentrations of C RGO and C MG (*** p< 0 00001) 5

Figure 5A

Figures 5A-5F show the size dist ri bution of graphene oxide (GO) (5A), C RGO (5 B), CRGO- COO H (5C), MG (5 D), C MG (5 E), and doxorubicin-CMG (5F)

Figure 6

Figure 6 shows representative results from ex vivo ﬂorescence images of organs (lung, liver, kidney, spleen, and prostate) from mice (BL₆ and transgenic adenocarcinoma of mouse prostate tumor bearing (TRAMP) mice) harvested 4h post intravenous (I V) in j ection of Cy 5 5-10 C MG nanoparticles or …

Figure 7

Figure 7 shows the average ﬂuorescence intensity of organs (lung, liver, kidney, spleen, and prostate) from mice (BL₆ and TRAMP) harvested 4h post I V in j ection of Cy 5 5-CMG nanoparticles or PBS (control) Fluorescence was normalized to the weight of each organ Error bars were based on six tumors …

Figure 8

Figure 8 shows representative ex vivo magnetic resonance (MR) i mages of tumor and liver tissue in j ected with C MG nanoparticles or PBS (control)

Figure 9

Figure 9 shows the mean signal intensity of tumors and kidneys after I V administration of C MG or PBS (** p< 0 01)

Figure 10A

Figures 10A-10 D show i ron dist ri bution as shown by Prussian blue staining of tumor 20 (10A and 10 B) and kidney (10 C and 10 D) tissue samples from a mouse that received CMG nanoparticles (10 B and 10 D) or PBS (10A and 10 C) as a control

Figure 11A

Figure 11 A shows the representative ﬂuorescence images from in j ected C₅₇BL/6 and TRAMP mice

Figure 11B

Figure 11 B shows average ﬂuorescence intensity of each organ from in j ected C₅₇BL/6 and TRAMP mice

Figure 11C

Figure 11 C shows the representative ﬂuorescence images from in j ected C₅₇BL/6 and Lewis lung 5 carcinoma 1 (LLC₁) Tumor bea ri ng mice

Figure 11D

Figure 11 D shows average ﬂuorescence intensity of each organ from in j ected C₅₇BL/6 and LLC₁ Tumor bea ri ng mice

Figure 12

Figure 12 shows UV-Vis absorption spectra of (a) C MG, (b) doxorubicin-CMG nanoparticles, and (c) doxorubicin alone

Figure 13

Figure 13 shows the eﬀect of initial doxorubicin concentration on the doxorubicin loading 10 capacity of CMG

Figure 14

Figure 14 shows the cumulative release of doxorubicin from doxorubicin-CMG nanoparticles at pH of about 7 4 and about 5 1 (**p < 0 0001)

Figure 15A

Figures 15A-15F show confocal microscopic images of A549cells after 20h incubation with doxorubicin (15A-15C) or doxorubicin-CMG nanoparticles (15D-15F) Subcellular 15 localization of doxorubicin (15B and 15 E) was demonstrated via a ﬂuorescence assay Nucle i were stained with DAPI as a control (15 …

Figure 16

Figure 16 shows the viability of A₅₄₉ cells treated with diﬀerent concentrations of doxorubicin and doxorubicin-CMG nanoparticles (*p < 0 05) 20

Figure 17

Figure 17 shows a representative gel electrophoresis of complexes of nanoparticles and DNA at diﬀerent weight rations

Figure 18A

Figures 18A-18F show phase contrast (18A and 18D), ﬂuorescence (18B and 18E), and merged phase contrast/ﬂuorescence (18 C -18F) microscopy images of C 42b cells 48 h post- transfection with C MG nanoparticles (18A-18 C) or LipofectamineT M reagent (18D-18F) 25 complexed with a red-ﬂuorescent …

Figure 19A

Figures 19A-19B show the transfection eﬃciency of C42b cells (19A) (p = 0 37)and A₅₄₉ cells (19B) (p = 0 10) Transfection eﬃciency was quantiﬁed by Image J program

Figure 20A

Figures 20A-20P show GFP expression (20 C, 20G, 20K, and 200), DAPI (20A, 20E, 201, and 20M), doxorubicin (20B, 20F, 20J, and 20N), as indicated by a ﬂuorescence assay, and 5 the merged image for compa ri son (20 D, 20H, 20L, 20P) of LLC₁ tumor cells implanted on the ﬂanks (left and ri ght) of mice …

Figure 21A

Figures 21A-21B show quantiﬁcation of GFP expression as imaged in

Claims

Claims define the patent's legal scope. Independent claims stand alone; dependent claims (nested) narrow them. Click a claim to expand its dependents.

15 independent · 5 dependent

1
Independentchemically reduced graphene sheetchitosanMRI contrast agentchemotherapeutic agentDNA plasmidtheranostic nanoparticle
A nanoparticle comprising: a chemically reduced graphene sheet; chitosan, wherein the chitosan is attached to or loaded onto the chemically reduced graphene sheet; a magnetic resonance imaging (MRI) contrast agent attached to or loaded onto the chemically reduced graphene sheet; a chemotheraputic agent attached to or loaded onto the chemically reduced graphene sheet; and a DNA plasmid attached to or loaded onto the chemically reduced graphene sheet.
1. 2
  Dependent← claim 1theranostic nanoparticle
  The nanoparticle of claim 1, wherein the nanoparticle is a theranostic agent.
2. 6
  Dependent← claim 1MRI contrast agent
  The nanoparticle of claim 1, wherein the MRI contrast agent is a radio contrast agent.
3. 11
  Dependent← claim 1targeting moietytheranostic nanoparticle
  The nanoparticle of claim 1, further comprising a targeting moiety attached to the chemically reduced graphene sheet.
3
Independent
3-4. canceled
4
Independent
canceled
1. 5
  Dependent← claim 4MRI contrast agent
  The nanoparticle of claim 4, wherein the MRI contrast agent is super paramagnetic iron oxide. -2-
7
Independent
-10. canceled
8
Independent
canceled
9
Independent
canceled
10
Independent
canceled
12
Independentchemically reduced graphene sheetchitosanMRI contrast agentchemotherapeutic agentDNA plasmidtheranostic nanoparticle
A theranostic formulation comprising: a nanoparticle comprising: a chemically reduced graphene sheet; chitosan, wherein the chitosan is attached to or loaded onto the chemically reduced graphene sheet; a magnetic resonance imaging (MRI) contrast agent attached to or loaded onto the chemically reduced graphene sheet; a chemotherapeutic agent attached to or loaded onto the chemically reduced graphene sheet; a DNA plasmid attached to or loaded onto the chemically reduced graphene sheet; and a pharmaceutically acceptable carrier.
13
Independent
-18. canceled
14
Independent
canceled
15
Independent
canceled
16
Independent
canceled
17
Independent
canceled
18
Independent
-3- canceled
19
Independentchemically reduced graphene sheetchitosanMRI contrast agentchemotherapeutic agentDNA plasmidgraphene oxide sheettheranostic nanoparticle
A method for generating theranostic nanoparticles com pris i ng: reducing a graphene oxide sheet to form chemically reduced graphene oxide sheet; loading or attaching a magnetic resonance imaging (MRI) contrast agent onto the chemically reduced graphene oxide sheet; loading or attaching chitosan onto the chemically reduced graphene oxide sheet; loading or attaching a chemotherapeutic agent onto the chemically reduced graphene oxide sheet; and loading or attaching a DNA plasmid moiety onto the chemically reduced graphene oxide sheet.
1. 20
  Dependent← claim 19targeting moietytheranostic nanoparticle
  The method of claim 19, further comprising the step of attaching or loading a targeting moiety onto the chemically reduced graphene oxide sheet. -4-

Worked examples

Embodiments described in the patent, grouped by the materials and process steps they use.

Doxorubicin-CMG-GFP DNA nanoparticle in vivo study (IV injection)

example section example

2 materials1 process step

LLC₁ tumor-bearing mice (n=2 per group) were injected with doxorubicin-CMG-GFP DNA nanoparticles by IV (25 µg DNA/mouse in 100 µL). At 24 or 48 h after injection, mice were sacrificed and frozen organ sections were examined for GFP expression and doxorubicin fluorescence in kidney, liver, lung, and spleen tissue.

Doxorubicin-CMG-GFP DNA nanoparticle in vivo study (IP injection)

example section example

2 materials

LLC₁ tumor-bearing mice (n=2 per group) received a single IP injection (25 µg DNA/mouse in 100 µL) of doxorubicin-CMG-GFP DNA nanoparticles. At 24 or 48 h after injection, mice were sacrificed and frozen organ sections were immunostained with anti-GFP antibody and counterstained with DAPI. Quantification of GFP expression and doxorubicin fluorescence was normalized to control background by ImageJ.

Device structures

Layer stacks claimed or described, ordered top of device to substrate.

theranostic nanoparticle

DNA plasmidgene therapy moiety (DNA plasmid)

chemotherapeutic agentdrug moiety (chemotherapeutic)

MRI contrast agentdiagnostic moiety (MRI contrast agent)

chitosanfunctionalization moiety

chemically reduced graphene sheetscaffold

Materials

Materials described outside the worked examples.

chemically reduced graphene sheet

Nanoparticle Scaffold/Carrier

chitosan

Functionalization Moiety/Biocompatibility Agent

Characterization

Measurements and analyses referenced in the patent, with their drawing references.

ftir ir

Ftir Ir

Figure 2 shows Fou ri ner Transform Infrared (FTIR) spectra of (a) chemically reduced 25 graphene oxide (C RGO), (b) CRGO- COO H, and (c) CMG 3 T| H Docket Number 292103-

Figure references

Figure 2

graphene

Reported properties

Performance values and ranges asserted in the specification or claims.

Property	Value	Material
Duration	169–173 Second	—
Thickness

Why these are connected

Related documents with shared materials, methods, properties, or citations.

GRAPHENE BASED THERANOSTICS FOR TUMOR TARGETED DRUG/GENE DELIVERY AND IMAGING | Matter42 Literature