Methods and compositions to enhance the immunogenicity of tumors

An immunogenic, tumor cell-based technology, applied in drug combinations, antineoplastic drugs, chemical instruments and methods, etc.

Pending Publication Date: 2020-06-19
INSIDEOUTBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite the progress, immunomodulatory approaches that specifically target tumor cells and enhance tumor immunogenicity are

Method used

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  • Methods and compositions to enhance the immunogenicity of tumors
  • Methods and compositions to enhance the immunogenicity of tumors
  • Methods and compositions to enhance the immunogenicity of tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Method for selecting immunoreactive peptides from C3d

[0067] The methods described herein and in Example 2 are based on the methods of Knopf, P.M. et al., Immunol. Cell Biol. (2008) 86, 221-225, and De Groot, A.S., Immunol. Cell Biol. (2014) 1-9, to Identification of immunostimulatory MHC class II binding peptides present in C3d. This method can be extended to identify other peptides suitable for use in the present invention. The purpose of these extensions is to:

[0068] 1. Identification of high-affinity class I and class II major histocompatibility complex (MHC) binding sites for C3d-derived peptides using a non-proprietary algorithm;

[0069] 2. Ensure that metalloproteases in the tumor environment can digest them to the appropriate size for binding to MHC proteins.

[0070] step

[0071] 1. Human C3d amino acid sequence: (SEQ ID NO: 5)

[0072]

[0073] Alignment with sequences from dog, pig, cow, mouse and rat to identify conserved regions. ...

Embodiment 2

[0090] Example 2: C3 MHC Peptides

[0091] Figure 6 Listed are the C3 wild-type amino acid sequences (SEQ ID NOs: 6, 8, 10, 12 and 14) and their associated mutant sequences (SEQ ID NOs: 7, 9, 11, 13 and 15), which have been Optimized to improve MHC binding and alter the proteolytic site of metalloproteases. The mutated peptides have reasonably improved MHC binding and improved immune stimulation.

Embodiment 3

[0092] Example 3: Design of Guide RNA Sequences for CRISPR

[0093] For the method of gene editing using CRISPR technology, a guide RNA sequence is required that edits / cuts out the C3 gene but not the C3d gene sequence properly. See eg portals.broadinstitute.org / gpp / public / analysis-tools / sgrna-design. Exemplary guide RNA sequences (SEQ ID NO: 16-18) and corresponding PAM sequences are at Figure 7 shown in .

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Abstract

Methods and compositions for enhancing the immunogenicity of a tumor of interest by modulating/altering the expression of specific complement proteins and complement protein receptors associated withthe immune suppression of a tumor/tumor cell are described herein.

Description

[0001] related applications [0002] Pursuant to 35 U.S.C. §119(e), this application claims the benefit of the following U.S. provisional application numbers: 62 / 556,836 filed September 11, 2017, 62 / 582,365 filed November 7, 2017, and April 12, 2018 62 / 656,495 filed on , the entire contents of which are hereby incorporated by reference in their entirety. [0003] Incorporate material into ASCII text files by reference [0004] This application contains a Sequence Listing, which has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy was created on September 10, 2018, named 0371_0001WO1_SL.txt, and was 22,560 bytes in size. Background technique [0005] Cancerous tumors form an invisible barrier to protect themselves from the body's immune-driven defense mechanisms. Various approaches have been developed to enhance the immune response to this tumor, including inoculation with bacterial infections, viruses, vaccin...

Claims

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Application Information

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IPC IPC(8): C12N15/113C07K14/47C07K14/705A61K38/17A61P35/00A61P37/04
CPCA61K38/1725C07K14/70596C07K14/472C12N15/113C07K2319/00C12N2310/14C12N2310/20A61K38/465A61K45/06A61K31/7105C12N15/907A61P35/00A61K2300/00A61K9/127A61K9/51A61K38/00
Inventor 艾伦·戈登·赫伯特
Owner INSIDEOUTBIO INC
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