Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Compositions and methods for treatment of cytomegalovirus

A virus-like, murine leukemia virus technology, applied in the field of virus envelope glycoprotein variants and virus-like particles, can solve problems such as toxicity and limited efficacy

Inactive Publication Date: 2015-01-07
VARIATION BIOTECHNOLOGIES INC
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing therapies are characterized by significant toxicity and limited efficacy, especially for late-onset disease (Boeckh M., 2004 Pediatr Transplant 8 (Suppl. 5): 19-27; Limaye AP., 2004 Transplantation 78 (9): 1390- 6), and they have no effect on congenital HCMV disease

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compositions and methods for treatment of cytomegalovirus
  • Compositions and methods for treatment of cytomegalovirus
  • Compositions and methods for treatment of cytomegalovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[1015] Embodiment 1: Construction of DNA expression plasmid

[1016] This example describes the development of expression plasmids and constructs for expression of recombinant HCMV gene sequences such as gB, gB-G, gH-G and Gag / pp65 fusion gene sequences. A standard expression plasmid usually consists of the following: mammalian-derived promoter sequence, intron sequence, polyadenylation signal sequence (PolyA), pUC origin of replication sequence (pUC-pBR322 is the colE1 origin of replication origin / site and antibiotic resistance genes for replication of plasmids in bacteria such as E. coli (DH5α) and as selectable markers for plasmid plaque selection. Following the introns in the plasmid are multiple restriction enzyme sites that can be used for splicing into a gene or partial gene sequence of interest.

[1017] Propol II expression plasmid containing pHCMV (early promoter for HCMV), β-globin intron (BGL intron), rabbit globin polyadenylation signal sequence (PolyA), pUC orig...

Embodiment 2

[1020] Embodiment 2: the production of virus-like particle (VLP)

[1021] This example describes methods for producing virus-like particles (VLPs) containing various recombinant HCMV antigens described in Example 1.

[1022] HEK293T cells (ATCC, CRL-11268) were transiently transfected with MMLV-Gag DNA expression plasmids using the calcium phosphate method and co-transfected with gB or gB-G (data not shown) or gH-G DNA expression plasmids. Alternatively, cells were transfected with Gag / pp65 DNA expression plasmids and co-transfected with gB or gB-G (data not shown) or gH-G DNA expression plasmids. It will be appreciated that cells can be transfected with MMLV-Gag DNA expression plasmids and co-transfected with gB and gH-G or both gB-G and gH-G DNA expression plasmids. Multiple HCMV antigens expressed by HEK293 cells were confirmed by flow cytometry ( figure 2 A). 48 to 72 hours after transfection, the VLP-containing supernatant was harvested and filtered through a 0.45 μm ...

Embodiment 3

[1023] Example 3: Physicochemical Characterization of Virus-Like Particles (VLPs)

[1024] Physicochemical analysis of VLPs included particle size determination and evaluation of polydispersity index using a Malvern Instrument Zeta-Sizer Nano series (ZEN3600). Exemplary results obtained from nanonization analysis are in image 3 A and 3B are shown. An exemplary VLP composition (gH-G / pp65 bivalent VLP composition) was produced in two different laboratories using the same recombinant expression vector and both VLP preparations gave average particle sizes ranging from 150 nm to 180 nm in diameter . This is consistent with the size of CMV virions, which are reported to be in the range of 150 nm to 200 nm (1997 J Pathol 182:273-281). A low polydispersity index (PdI) of 0.214 to 0.240 indicates product homogeneity and narrow size distribution.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
particle sizeaaaaaaaaaa
polydispersity indexaaaaaaaaaa
Login to View More

Abstract

The present disclosure provides compositions and methods useful for treating HCMV infection. As described herein, the compositions and methods are based on development of immunogenic compositions that include virus-like particles (VLPs) which comprise one or more Moloney Murine leukemia virus (MMLV) core proteins and include one or more HCMV epitopes, such as, for example, from HCMV envelope glycoproteins gB and / or gH and / or tegument protein pp65. Among other things, the present invention encompasses the recognition that a combination of antigens (e.g., envelope glycoproteins and structural proteins) can lead to beneficial immune responses, for example that include both a humoral response (e.g., production of neutralizing antibodies) and a cellular response (e.g., T-cell activation).

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Application No. 61 / 558,800, filed November 11, 2011, and U.S. Provisional Application No. 61 / 654,157, filed June 1, 2012, the contents of which are hereby incorporated by reference in their entirety . Background technique [0003] Human cytomegalovirus (HCMV) (beta-herpesvirus) is a ubiquitous pathogen. HCMV infection is usually overlooked in immunocompetent persons, with the mildest and nonspecific symptoms. In contrast, certain high-risk groups, such as in immunosuppressed patients (such as AIDS patients or transplant recipients), and after prenatal infection, HCMV infection has a severe manifestation (Staras SA et al., 2006 Clin Infect Dis43(9): 1143-51; Hebart H et al., 2004 Hum Immunol 65(5):432-6; Rowshani AT et al., 2005 Transplantation 79(4):381-6). Existing treatments include the use of immunoglobulins and antiviral agents (such as ganciclovir and its deriv...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63A61K39/245A61K39/295A61P31/22A61P37/04C07K14/045C07K14/145C07K14/15C07K19/00C12N15/38C12N15/48C12N15/62C12N15/86C12N7/01C12N7/04
CPCC07K2319/00A61K39/245C12N2740/13023A61K39/12A61K2039/5258C07K2319/03C07K14/005C12N2710/16134C12N2760/20234A61K2039/70A61P31/22C12N2760/20245A61P31/20
Inventor 大卫·E·安德森安妮-凯瑟琳·弗卢基格
Owner VARIATION BIOTECHNOLOGIES INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com