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Methods and compositions for improved delivery, expression or activity of RNA interference agents

A technology of RNA interference, composition, applied in the direction of DNA/RNA fragments, recombinant DNA technology, botanical equipment and methods, etc.

Inactive Publication Date: 2012-10-03
杰西．L．S．奥 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(6) DNA / lipid complexes or DNA / polymer complexes that cannot be separated from the content may be degraded in lysosomes

Method used

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  • Methods and compositions for improved delivery, expression or activity of RNA interference agents
  • Methods and compositions for improved delivery, expression or activity of RNA interference agents
  • Methods and compositions for improved delivery, expression or activity of RNA interference agents

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0204] Preparation of RNAi lipid carrier

[0205]Cationic liposomes were prepared as follows. The desired weight of each lipid was mixed and dissolved in a 90:10 volume ratio mixture of chloroform and methanol. To prepare Pac-PCat-RNAi, add the required amount of paclitaxel to the dissolved lipid. Paclitaxel is added in an amount of 0.04% by weight of the total lipid weight. In the culture medium of a typical in vitro experiment, 0.04% paclitaxel by weight provides a total of 10 nM equivalent concentration of paclitaxel, while 0.2% paclitaxel by weight provides an equivalent concentration of 50 nM. For Doc-PCat-RNAi, Col-PCat-RNAi and Vin-PCat-RNAi, then add appropriate amount of docetaxel, colchicine and vincristine to provide the equivalent concentration of the culture medium as docetaxel 10nM, colchicine 100nM , vincristine 10 nM. For a typical total lipid of 10 mg, 5 mL of chloroform / methanol is required. The dissolved lipids were placed in a round bottom flask and ...

example 2

[0208] RIDES plus tumor loosening is effective in vivo: intraperitoneal administration for intraperitoneal tumors

[0209] Experiments were performed in immunosuppressed mice bearing intraperitoneal HS766T pancreatic cancer. Tumor cells were injected into the peritoneal cavity of mice, resulting in tumor formation throughout the peritoneal cavity. In the absence of treatment, the established tumor models eventually resulted in the death of 100% of the animals. Treatment was initiated 10 days after tumor implantation, or half the expected survival time of untreated animals. All treatments were administered intraperitoneally. Day zero is the first treatment and survival time is the time since the first treatment. Paclitaxel was administered in the form of tumor penetrating particles (TPM), prepared as described in US patent application Ser. No. 11 / 242,546. Animals were given a single dose of TPM (immediate-release and sustained-release granules containing a total equivalen...

example 3

[0219] RIDES works in vivo: intravenous therapy for subcutaneous tumors

[0220] This example demonstrates that intravenously administered RIDES is effective against systemic tumors. The study was performed in immunosuppressed mice bearing subcutaneously implanted human xenograft tumors. Three different types of tumors, namely, pancreatic cancer HS766T, prostate cancer PC3, pharyngeal cancer FaDu. Treatment begins when tumors reach a size greater than 3 mm. Paclitaxel dissolved in polyoxyethylene castor oil 50:50 V / V in ethanol. RNAi is survivin siRNA. The RNAi carrier was PCat liposome, loaded with 1 nmol of siRNA against wild-type survivin mRNA, and 0.12 mg DOTAP per animal. Day zero is the first treatment and survival time is the time from the first treatment.

[0221] Mice bearing HS766T tumors were randomly divided into six groups according to tumor size. Each group consisted of 4 or 5 animals. Treatment begins when tumors reach a size of about 3 mm in diameter...

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Abstract

The present disclosure provides methods and compositions for enhanced delivery of siRNA or miRNA, into the interior of multilayered tissues, and into the cytoplasm or nucleus of cells of a tissue. Such methods and compositions yield tumor-selective and intracellular delivery of RNAi agents and allow for RNAi-mediated activity such as knock-down of the target genes and associated products. The current disclosure further provides methods and compositions for improving the intracellular bioavailability of nucleotide agents.

Description

[0001] Cross References to Related Applications [0002] This application claims Provisional Patent Application Serial No. 61 / 250,587 filed October 12, 2009 and entitled "Methods and Compositions for Improved Management and Treatment of Patients with Tumors or Proliferative Disorders" and March 8, 2010 The benefit of an application filed under Serial No. 12 / 719,546 entitled "Methods and Compositions for Improved Delivery, Expression or Activity of RNA Interference Agents". The entire contents of these applications are incorporated herein by reference. The contents of all patents, patent applications, and references cited in this application are hereby incorporated by reference in their entirety. [0003] Statement Regarding Federally Funded Research [0004] This work was supported by a grant from the United States Department of Health and Human Services (grant number R43CA134047). technical field [0005] RNA interference (RNAi) can be used to correct the expression of a c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N25/26C12N15/11
CPCC12N2320/32C12N2310/14C12N2320/31C12N15/111
Inventor 杰西.L.S.奥M.吉尔劳姆.温特杰斯吕泽王杰
Owner 杰西．L．S．奥
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