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 invention provides temperature-sensitive compositions for stimulating an immune response against an antigen in a mammalian subject and for expressing a protein in the mammalian subject. The composition comprises an excipient and is administered by intradermal injection, and the ts-agent encodes the antigen or protein and is capable of expressing it at skin temperature of the subject but not at core temperature of the subject.

In particular embodiments, the ts-agent is a temperature-sensitive viral vector, including temperature-sensitive Sendai virus vectors, or a temperature-sensitive viral vector encoding a spike protein or fragment thereof of a coronavirus. In other embodiments, the ts-agent is a temperature-sensitive self-replicating RNA comprising a viral replicon lacking a viral structural protein coding region, where the RNA is not packaged in a viral particle.

For certain RNA designs, the ts-agent includes an Alphavirus replicon having a nonstructural protein coding region with an insertion of 12-18 nucleotides resulting in expression of a nonstructural Protein 2 (nsP2) comprising additional amino acids between beta sheet 5 and beta sheet 6. The concept is applied to stimulating an immune response against an antigen and to expressing a protein in a mammalian subject by intradermal injection.

Claims Coverage

The independent claims cover intradermal administration of excipient plus a temperature-sensitive agent encoding an antigen or protein, where expression occurs at skin temperature but not at core temperature. Across the independent claims, the key inventive features include temperature-dependent expression selectivity, temperature-sensitive viral vector or temperature-sensitive self-replicating RNA replicon architecture not packaged in viral particles, coronavirus spike or RBD as the antigen in some embodiments, and Alphavirus replicon nsP2 insertion features.

Collectively, the independent claims require intradermal injection of an excipient-containing composition with a temperature-sensitive agent that encodes the antigen or protein and expresses it at skin temperature but not at core temperature, using either a temperature-sensitive viral vector or a temperature-sensitive self-replicating RNA replicon not packaged in viral particles. Further independent-claim specificity includes coronavirus spike, including RBD, as the antigen and, in one embodiment, an Alphavirus replicon with a defined nsP2 insertion architecture.

Stated Advantages

Documented Applications