Modified factor ix, and compositions, methods and uses for gene transfer to cells, organs, and tissues

a technology of factor ix and fusion factor, applied in the field of modified, can solve the problems of need for repeated infusion, cost of treatment, risk of developing anti-therapeutic factor immune responses,

Inactive Publication Date: 2020-04-30
THE CHILDRENS HOSPITAL OF PHILADELPHIA
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0165]Methods and uses of the invention include treatment methods, which result in any therapeutic or beneficial effect. In various invention methods and uses, further included are inhibiting, decreasing or reducing one or more adverse (e.g., physical) symptoms, disorders, illnesses, diseases or complications caused by or associated with the disease. For a bleeding disorder such as hemophilia, a therapeutic or beneficial effect includes, but is not limited to, reduced bruising, reduced blood clotting time, reduced bleeding episodes (duration, severity, frequency). For example, reduced duration, severity or frequency of joint or cerebral (brain) bleeding episodes. For a bleeding disorder such as hemophilia, a therapeutic or beneficial effect also includes, but is not limited to, reduced dosage of a supplemental clotting factor protein (e.g., Factor IX protein) or elimination of administration of a supplemental clotting factor protein (e.g., Factor IX protein).

Problems solved by technology

However, this therapeutic approach has several drawbacks such as the need for repeated infusions, the cost of the treatment, the risk of developing anti-therapeutic factor immune responses, and the risk of potentially fatal bleedings.

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
  • Modified factor ix, and compositions, methods and uses for gene transfer to cells, organs, and tissues
  • Modified factor ix, and compositions, methods and uses for gene transfer to cells, organs, and tissues
  • Modified factor ix, and compositions, methods and uses for gene transfer to cells, organs, and tissues

Examples

Experimental program
Comparison scheme
Effect test

example 1

sign / Preparation

[0233]A novel Factor IX nucleic acid encoding a high specific activity human factor IX protein having the 338 L Padua variant (Simioni P, et al., N Engl J Med 2009, 361:1671) was designed (“FIX 39-Padua”; SEQ ID No: 10; FIG. 10). FIX39-Padua is completely devoid of CpG dinucleotides in the FIX coding and intronic sequences. For comparative testing, FIX19 (Mingozzi et al. Sci. Transl Med. 2013) was prepared and modified to include the FIX Padua, to rule out any potential confounding effects resulting from the FIX Padua (“FIX 19-Padua”; SEQ ID NO:11; FIG. 11).

[0234]A plasmid (“pAAV-ApoE_hAAT-FIX39”; 11125 bp; SEQ ID NO: 12; FIG. 12A) was synthesized and included the FIX39-Padua expression cassette and the elements described in Table 2. A map of pAAV-ApoE_hAAT-FIX39 is shown in FIG. 13.

TABLE 2pAAV-ApoE_hAAT-FIX395′ AAV2 ITRSEQ ID NO: 13Enhancer (HepaticSEQ ID NO: 14Control Region)hAAT promoterSEQ ID NO: 155′ UTRSEQ ID NO: 16FIX39-Padua CDSSEQ ID NO: 10(FIG. 10)Intron AS...

example 2

AAV Variant 4-1 Transduction

[0237]Primary hepatocytes from cynomolgus macaque and human origin were transduced with the 4-1 variant capsid (SEQ ID NO:4) expressing luciferase at four different multiplicities of infection (MOI) ranging from 500 to 62,500 vector genomes per cell. Seventy-two hours after transduction, luciferase expression was analyzed. As shown in FIG. 16, the ratio of transduced human hepatocytes relative to non-human primate hepatocytes ranged from 0.8 to 1.5, depending on the MOI used. These data generated in vitro appear to be consistent with previous observations in vivo when comparing expression of coagulation factor IX in cynomolgus macaques and human subjects.

example 3

tudy

[0238]A study was conducted to evaluate the potency of AAV-FIX39-Padua versus AAV-FIX19-Padua in mice. Groups of 5 mice were injected at 8-10 weeks of age with either 1×1011 or 1×1012 vg / kg of AAV-FIX39-Padua and AAV-FIX19-Padua. Following vector administration, blood was collected by retro-orbital bleeding using heparinized capillary tubes; plasma was isolated by centrifugation at 9000 rpm for 10 minutes at 4° C. and stored frozen at −80° C. until assayed.

[0239]Plasma collected was used to evaluate hFIX transgene expression. Human FIX levels in plasma were measured using an ELISA kit (Affinity Biologicals, Ancaster, ON, Canada).

[0240]Activity levels of human FIX were measured by activated partial thromboplastin time (aPTT) assay. The aPTT assay was performed by mixing sample plasma in a 1:1:1 volume-ratio with human FIX-deficient plasma (George King Biomedical, Inc) and aPTT reagent (Trinity Biotech), followed by a 180s incubation period at 37° C. Coagulation was initiated by a...

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
pHaaaaaaaaaa
nucleic acidaaaaaaaaaa
nucleic acid sequenceaaaaaaaaaa
Login to view more

Abstract

The invention relates to modified Factor IX coding sequence, expression cassette, vectors such as viral (e.g., lenti- or adeno-associated viral) vectors, and gene transfer methods and uses. In particular, to target Factor IX nucleic acid to cells, tissues or organs for expression (transcription) of Factor IX.

Description

RELATED APPLICATIONS[0001]This patent application is a continuation application of U.S. application Ser. No. 15 / 191,357, filed Jun. 23, 2016, which claims the benefit of priority to provisional application No. 62 / 183,599, filed Jun. 23, 2015, application No. 62 / 315,453, filed Mar. 30, 2016, application No. 62 / 338,315, filed May 18, 2016, application No. 62 / 348,781, filed Jun. 10, 2016 and application No. 62 / 349,572, filed Jun. 13, 2016. The entire contents of the foregoing applications are incorporated herein by reference, including all text, tables, sequence listing and drawings.SEQUENCE LISTING[0002]The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Dec. 5, 2019, is named “CHOP0506152_ST25.txt” and is 26.4 KB in size.INTRODUCTION[0003]Genetic disorders, caused by absence or a defect in a desirable gene (loss of function) or expression of an ...

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(United States)
IPC IPC(8): A61K38/48C12N9/64C12N15/86
CPCC12Y304/21022C12N2750/14143A61K38/4846C12N9/644C12N15/86A61P7/04A61K48/00C07K14/745C12N15/52C12N15/8645C12N2750/14132C12N2750/14141
Inventor HIGH, KATHERINE A.ANGUELA, XAVIER
Owner THE CHILDRENS HOSPITAL OF PHILADELPHIA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap