Temperature-based transient delivery of nucleic acids and proteins to cells and tissues

Inventors

Ko, Minoru S. H.

Assignees

Elixirgen Therapeutics Inc

Abstract

The present disclosure relates to methods for transiently activating temperature-sensitive agents in one or more cells, for example by contacting one or more cells with a temperature-sensitive agent and transiently incubating the cells at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Additionally, the present disclosure relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent and then lowering the subject's core body temperature to a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. The disclosure also relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent, maintaining the subject's surface body temperature at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Further disclosed are methods of treating a subject with a temperature-sensitive therapeutic agent.

Core Innovation

The document describes temperature-based transient activation of temperature-sensitive agents (ts-agents) in which a ts-agent expresses an antigen at a permissive temperature but not at a non-permissive temperature. The approach defines ts-agents as capable of being active only within a permissive temperature range and inactive at a non-permissive temperature, enabling control of expression by temperature.

For immune stimulation, the document provides ts-agent compositions comprising an excipient and a ts-agent encoding an antigen, where the antigen is a spike protein or fragment thereof of a coronavirus. The document specifically includes temperature-sensitive Sendai viral vector embodiments and temperature-sensitive self-replicating Alphavirus replicon RNA embodiments lacking viral structural protein coding regions, with expression behavior tied to permissive versus non-permissive temperature.

The document further characterizes ts-agents as including temperature-sensitive self-replicating RNAs and temperature-sensitive polypeptides. It also describes a temperature-sensitive self-replicating Alphavirus-based RNA design in which a nonstructural protein coding region contains an insertion of 12–18 nucleotides, resulting in expression of nonstructural protein 2 (nsP2) comprising 4 to 6 additional amino acids between beta sheet 5 and beta sheet 6 of nsP2.

Claims Coverage

The partial content identifies three independent claims. Across these claims, the inventive subject matter is organized around temperature-sensitive expression behavior at permissive versus non-permissive temperatures, temperature-sensitive agent type, and specific sequence/structural modifications to nsP2.

Overall, the independent claim coverage ties transient activity to ts-agents that express at a permissive temperature and not at a non-permissive temperature, with immune-response compositions using either a temperature-sensitive Sendai viral vector or a temperature-sensitive self-replicating Alphavirus replicon RNA lacking viral structural protein coding regions, and a separate agent-structure claim specifying a 12–18 nucleotide insertion that alters nsP2 between beta sheet 5 and beta sheet 6.

Stated Advantages

Documented Applications