METHODS AND COMPOSITIONS FOR RNAi-BASED CANCER TREATMENT

a cancer treatment and composition technology, applied in the field of nucleotide-based agent-based and rnai-based cancer treatment methods and compositions, can solve the problems of slow pc disease metastasis and epithelial-to-mesenchymal transition, and achieve the effect of reducing the levels of downstream effector proteins

Inactive Publication Date: 2014-10-23
SILENSEED LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, anti-mutated K-ras treatment potentially can slow the epithelial-to-mesenchymal transition (EMT) and thereby slow metastasis of PC disease.

Method used

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  • METHODS AND COMPOSITIONS FOR RNAi-BASED CANCER TREATMENT
  • METHODS AND COMPOSITIONS FOR RNAi-BASED CANCER TREATMENT
  • METHODS AND COMPOSITIONS FOR RNAi-BASED CANCER TREATMENT

Examples

Experimental program
Comparison scheme
Effect test

example 1

Schematic Depiction of the Described Treatment

[0160]FIG. 1 shows a schematic depiction of the described treatment of metastatic cancer. Local and prolonged delivery of RNAi-based treatment within a primary tumor is administered, often in parallel to systemic administration of drugs including chemotherapy drugs

example 2

LODER-Derived siG12D Significantly Inhibits Growth of Pancreatic Cancer Cells In Vitro

Methods

[0161]DDD's containing RNAi molecules were produced in a biological-class hood in a clean room, as follows:

[0162]Step 1: Preparation of siRNA / D-Mannitol / Sodium Bicarbonate Mixture:

[0163]siRNA was added to the pre-weighed D-Mannitol and Sodium Bicarbonate, and they were dissolved in RNase-free sterile water.

[0164]Step 2: Freezing:

[0165]The liquid was placed into glass vials, frozen in dry ice, and lyophilized for 48 hours.

[0166]Step 3: PLGA Preparation:

[0167]PLGA was dissolved in Ethyl Acetate.

[0168]Step 4: Combining PLGA with D-Mannitol / Sodium Bicarbonate / siRNA:

[0169]The PLGA solution was poured into the glass vial containing the lyophilized D-Mannitol / Sodium Bicarbonate / siRNA in fractions and stirred until homogenization

[0170]Step 5: Solvent Evaporation.

[0171]The solution was poured into a Teflon-covered dedicated glass dish and left to evaporate inside a dedicated container for 3-5 days.

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example 3

Functionality of the siRNA LODER In Vivo

[0181]For the in vivo assessment of LODER-siRNA effects, we initially used the murine colon cancer CT26 cells. These cells were stably transfected to constitutively express the luciferase gene (CT26-LUC cells). CT26-LUC cells were injected subcutaneously into BALB / c mice. When tumor volume reached ˜1 cm3, two siLuc or control (siGFP) LODERs were implanted into the tumors. As seen in FIG. 3A, luciferase expression increased by approximately 3-fold in the siGFP LODER-implanted mice, while in mice implanted with the siLuc LODERs, the luciferase levels barely increased. FIG. 3B reveals that there was no significant effect on tumor weight, at sacrifice, in the group treated with the siLuc LODER in comparison with the siGFP LODER group, indicating that the lower Luciferase levels were not caused by an antitumor effect; rather, the siLuc released from the LODER inhibited luciferase expression, as measured by its activity in vivo.

[0182]We also assesse...

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Abstract

The present invention generally concerns methods and nucleotide-based compositions for treating metastatic cancer that is associated with a primary tumor or a few tumors at the primary organ.

Description

[0001]This application is a Continuation-in-Part of International Application No. PCT / IL2013 / 050340, filed on Apr. 18, 2013, which claims priority from U.S. patent application Ser. No. 13 / 451,231, filed on Apr. 19, 2012. The contents of each of these applications are incorporated herein by reference.FIELD OF THE INVENTION[0002]The present invention generally concerns nucleotide-based agent-based and RNAi-based methods and compositions for treating metastatic cancer.BACKGROUND OF THE INVENTION[0003]Metastasis is the spread of a cancer from one organ, usually the primary tumor, to another non-adjacent organ. Typically, pancreatic cancer first metastasizes to regional lymph nodes, and later to the liver or to the peritoneal cavity and, rarely, to the lungs, bone or brain. Metastases represent the end-products of a multi-step cell-biological process (Valastyan and Weinberg 2011) termed the invasion metastasis cascade, which involves dissemination of cancer cells to anatomically distant ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/113
CPCC12N15/113A61K31/713A61K31/7105A61K31/7115A61K9/0024A61K47/34C12N15/1135C12N2310/14A61P35/00A61P35/04C12N2310/11
Inventor SHEMI, AMOTZKHVALEVSKY, ELINA ZORDE
Owner SILENSEED LTD
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