NUCLEIC ACID MOLECULES AND USES THEREOF
Provided in this application are formulations of double stranded RNA molecules and Krebs Cycle analogs that improving ribonuclease stability, reducing off-target effects of a double stranded siRNA molecule, or of reducing interferon responsiveness of a double stranded siRNA molecule using such dsRNA. Also disclosed are methods of treating a primary tumor or a metastasis by contacting circulating tumor cells, a primary tumor, or a metastasis with a described formulation.
1 . A formulation, comprising:
(a) an RNAi molecule comprising at least on: locked nucleic acid (LNA), unlocked nucleic acid (UNA), bridged nucleic acid (BNA), glycerol nucleic acid (GNA), or a combination thereof; and
(b) an RNAi carrier.
2 . The formulation of claim 1 , wherein the carrier provides for one or more of the following: stability for shortened duplexes, reduction or prevention of sense strand loading, reduction or prevention of seed region microRNA adverse side effects and reduction of non-specific immunoactivation.
3 . The formulation of claim 1 , wherein the RNAi carrier is a di-lipid amino acid (DILA 2 ).
4 . The formulation of claim 1 , wherein the RNAi carrier is a Krebs Cycle analog.
5 . The formulation of claim 1 , wherein the RNAi carrier is a Krebs Cycle analog and wherein the Krebs Cycle analog reduces or prevents cytotoxicity.
6 . A formulation, comprising:
(a) an RNAi molecule; and
(b) a Krebs Cycle analog RNAi carrier.
7 . The formulation of claim 6 , wherein the RNA or RNA analog comprises a locked nucleic acid (LNA), an unlocked nucleic acid (UNA), a bridged nucleic acid (BNA), a glycerol nucleic acid (GNA), or a combination thereof.
8 . A formulation, comprising:
(a) an RNAi molecule comprising at least on: locked nucleic acid (LNA), unlocked nucleic acid (UNA), bridged nucleic acid (BNA), glycerol nucleic acid (GNA), or a combination thereof; and
(b) a Krebs Cycle analog RNAi carrier.
9 . A formulation, comprising:
(a) an RNAi molecule comprising at least on glycerol nucleic acid (GNA), or a combination thereof a glycerol nucleic acid (GNA) analog; and
(b) a Krebs Cycle analog RNAi carrier.
10 . Use of a formulation of any of claims 1 - 9 for the manufacture of a medicament for the treatment of cancer.
11 . Use of a formulation of any of claims 1 - 9 for the manufacture of a medicament for inducing apoptosis of a circulating tumor cell (CTC).
12 . Use of a formulation of any of claims 1 - 9 for the treatment of cancer.
13 . The use of claim 12 , wherein the cancer is characterized by the presence of a primary tumor or a metastasis.
14 . The use of claim 12 , wherein the cancer is breast cancer, a gastrointestinal cancer (such as a colon cancer), lung cancer or prostate cancer.
15 . The use of claim 12 , wherein the formulation is administered before, during, or immediately after surgery to remove a primary tumor or a metastasis.
16 . The use of claim 15 , wherein the formulation is locally administered at the site of the surgery.
17 . The use of claim 12 , wherein the formulation is administered in a time-release formulation.
18 . The use of claim 12 , wherein the formulation is administered by intravenous injection.
19 . The use of claim 12 , wherein the formulation exhibits reduced lipid-induced hepatic toxicity.
20 . The use of claim 12 , wherein the formulation reduces spread of the primary tumor or metastases.
21 . Use of a formulation of any of claims 1 - 9 for inducing apoptosis of a circulating tumor cell (CTC).
22 . The use of claim 21 , wherein the circulating tumor cell (CTC) is from a primary tumor or a metastasis.
23 . The use of claim 21 , wherein the formulation is administered in a time-release formulation.
24 . The use of claim 21 , wherein the formulation is administered by intravenous injection.
25 . The use of claim 21 , wherein the formulation is administered in a time-release formulation and by intravenous injection.
26 . The use of claim 21 , wherein the formulation exhibits reduced lipid-induced hepatic toxicity.
27 . Use of a formulation of any of claims 1 - 9 for inhibiting cancerous and pre-cancerous gene expression of breast cancer-related genes and pre-cancerous-related genes.
28 . The use of claim 27 , wherein the formulation is administered to an individual presenting with premalignant or malignant breast duct epithelial cells in a breast duct.
29 . The use of claim 27 , wherein the formulation is administered locally the breast duct.
30 . The use of claim 27 , wherein the formulation is administered in a time-release formulation.