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Conjugated virus-like particles and uses thereof as Anti-tumor immune redirectors

A virus-like, conjugated technology, applied in the direction of anti-tumor drugs, viral antigen components, viruses, etc., can solve the problems of high cost development channels and high medical expenses

Pending Publication Date: 2021-09-14
维伊木恩股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] As a result, current treatments retain the same well-recognized toxicity limitations, limited responder populations (not applicable to many cancers), and most importantly, they also have cost-prohibitive development pathways that result in patient healthcare costs higher

Method used

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  • Conjugated virus-like particles and uses thereof as Anti-tumor immune redirectors
  • Conjugated virus-like particles and uses thereof as Anti-tumor immune redirectors
  • Conjugated virus-like particles and uses thereof as Anti-tumor immune redirectors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0177] Conjugated VLPs bind to mouse tumors in vivo

[0178] The ability of the described conjugated VLPs to bind tumors is an important first step in the proposed mechanism of action (see figure 1 ). In vivo binding of VLPs was assessed using the murine cervical carcinoma model TC-1. Briefly, unconjugated VLPs were expressed in Expi293F TM Expi293F was used in human cells (ThermoFisher Scientific, Waltham, MA, US) TM produced by the expression system. In this protocol, 360 × 10 6 cells. Afterwards, to express HPV16L1 VLPs, cells were transfected with HPV16 L1 gene expression vector (Catalog No. 89910, Addgene, Cambridge, MA, US) via the manufacturer's protocol (ThermoFisher Scientific, Waltham, MA, US). Transfected cells were maintained for 72 hours before harvesting. HPV particles were released from Expi293F cells by detergent lysis, and the lysates were incubated overnight at 37°C with mild detergents such as Brij58 or Triton X-100. VLPs from overnight incubation ly...

Embodiment 2

[0184] Conjugated VLP binding to human tumors in vitro

[0185] To test the ability of conjugated VLPs to bind to human tumors in vitro, five human tumor cell lines will be evaluated: OVCAR3, MDA-MB231, HCT116, PC-3 and SKOV3 (ATCC, Manassas, VA, US). Each cell line was first seeded in one well of a 6-well plate. After 16-24 hours, cells will be dissociated using 0.05% trypsin. Cells were then counted to obtain 1 x 10 6 cells, centrifuged, and the supernatant removed, then resuspended in 100 μL of FACS buffer (1% FBS in PBS). Next, 0.3 μg / mL of conjugated VLPs was added to the respective cultures in vitro, and these cultures were incubated for 24 hours under optimal growth conditions. To demonstrate that the binding is HSPG specific, conjugated VLPs will or will not be pre-incubated with heparin (final concentration 0.1 mg / ml) and incubated for 1 hour at 4°C (to prevent endocytosis). After 1 hour of binding, the cells will be washed twice with 1 mL of FACS buffer before be...

Embodiment 3

[0187] Conjugated VLPs bind to human tumors in vivo

[0188] The described conjugated VLPs will be further tested to assess their ability to bind human tumors in vivo. In the first experiment, mice will be injected with OVCAR3 tumor cells (3×10 6 cells). Approximately two weeks after injection, the average tumor volume is expected to be approximately 10mm 3 . Three weeks after injection, Cu 2+ or Zn 2+Radiolabeled VLPs. VLP biodistribution and tumor specificity will be assessed at 4 hours, 12 hours, 24 hours and 48 hours. Tumors were then removed for weighing, radioactivity counting, viral microdistribution, and cell colocalization. In addition to tumors, imaging methods as well as histology will be used to assess the presence of VLPs in various major organs (such as liver, spleen, kidney) to assess the efficacy of VLP homing to tumors. In further experiments, mice will be injected with other tumor cell lines, such as HCT116, PC-3 or SKOV3, and the same experimental pr...

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Abstract

Disclosed is a new class of conjugated virus-like particles (VLPs), These conjugated VLPs bind a wide variety of tumors and comprise epitopes recognized by a prior T cell immune response already existing in a host. These epitopes are derived from pathogens or previous vaccinations (such as early childhood vaccines). This provokes the body's pre-existing cytotoxic immunity obtained through previous infection or previous childhood vaccination to be redirected to the tumor cells for tire elimination of cancer, and form long-term anti-tumor immunity. The described conjugated VLPs are useful for tailoring a broad range of tumors towards a response from existing immunity circumventing the need to identify tumor antigens or generate tumor-specific immune responses, importantly, the compositions and methods described herein broadens opportunities for treatment for ail cancer types in subjects who previously had un-targetable cancers due to various technological and biological limitations of currently available immuno-therapeutic drugs.

Description

[0001] priority [0002] This application claims priority to and benefit of U.S. Provisional Patent Application Serial No. 62 / 785,502, filed December 27, 2018, which is hereby incorporated by reference in its entirety. [0003] Reference to Sequence Listing submitted via EFS-WEB [0004] According to the EFS-Web legal framework and 37 CFR §§1.821-825 (cf. MPEP §2442.03(a)), an ASCII-compatible text file (named "8002PCT_ST25.txt", created on December 26, 2019, size 28,572 A sequence listing in the form of bytes) was filed at the same time as this application, and the entire content of the sequence listing is incorporated herein by reference in its entirety. technical field [0005] The present invention relates to conjugated virus-like particles (VLPs) that preferentially bind tumors. The VLPs of the invention include a recall protein capable of eliciting a T cell response using pre-existing recall protein-specific T cells that selectively attack the tumor to which the VLP bi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/025C07K19/00A61K39/00A61K39/02A61K35/17A61K35/76A61K35/768A61K39/12A61K38/16A61K39/39A61K47/64A61P35/00
CPCA61K9/107A61K35/17A61K2039/5258A61K2039/585A61K39/39A61K2039/545A61K39/02A61K39/12C07K14/005C12N2710/20023C12N2710/20022C12N2710/20033A61K35/768A61K35/76A61K38/162Y02A50/30A61P35/00C12N7/00G01N33/5094A61K39/00C12N2710/20032A61K2039/5256
Inventor J·W·王N·英加瓦K·松井
Owner 维伊木恩股份有限公司
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