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Compositions useful in treatment of metachromatic leukodystrophy

A malnutrition and white matter technology that can be used in drug combinations, gene therapy, peptide/protein components, etc., to solve problems such as disappointing results

Pending Publication Date: 2022-02-18
THE TRUSTEES OF THE UNIV OF PENNSYLVANIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, together with IT administered ERT in early-onset and late juvenile MLD (NCT01510028), the results in advanced infantile MLD patients with IV administered ERT were disappointing (NCT00418561)

Method used

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  • Compositions useful in treatment of metachromatic leukodystrophy
  • Compositions useful in treatment of metachromatic leukodystrophy
  • Compositions useful in treatment of metachromatic leukodystrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0275] Example 1 - AAV.hARSAco vector

[0276] The components of AAV.hARSAco are described in the table below.

[0277]

[0278] The vector was constructed from a cis-plasmid containing the coding sequence of human ARSA (SEQ ID NO:1 and SEQ ID NO:3) from chicken β with the cytomegalovirus enhancer flanked by AAV2 inverted terminal repeats. Actin promoter (CB7; SEQ ID NO: 16) expression.

[0279] Vectors were packaged in AAV serotype hu68 capsids (WO 2018 / 160582) by triple transfection of adherent HEK293 cells, and as described in Lock, M. et al., Rapid, Simple, and Versatile Manufacturing of Recombinant Adeno-Associated Viral Vectors at Purification was performed by iodixanol gradient centrifugation as previously described in Scale. Human Gene Therapy 21, 1259-1271 (2010).

[0280] More specifically, AAV.CB7.CI.hARSAco.rBG was produced by triple transfection of HEK293 Working Cell Bank (WCB) cells with the following triplet plasmid: AAV cis plasmid (pENN.AAV.CB7.CI.hARSAc...

Embodiment 2

[0350] Example 2 - Proof-of-Concept Pharmacology and Dose Ranging Studies in Mice

[0351] Experiments were performed to determine the efficacy of the AAVhu68.hARSAco vector following ICV injection in mice. Healthy 6-8 week old C57BL6 / J mice were selected for this proof-of-concept (POC) experiment. The ICV route was chosen because the small size of mice made it difficult to reliably inject AAV vectors via the ICM. Age was chosen based on historical data and our previous experience performing ICV injections in mice of this age (Hinderer et al., 2016). A necropsy time point of 21 days was chosen to capture stable transgene expression based on previous experience with other ICV-administered AAV vectors in mice (Hinderer et al., 2016).

[0352] at 1.00x10 10 Low dose for GC or 1.00x10 11High dose of GC, AAVhu68.CB7.CI.hARSAco.rBG vector was administered into the CSF of C57BL6 / J wild-type mice via a single ICV injection into the right ventricle. Doses were chosen based on prev...

Embodiment 3

[0356] Example 3 - Cell tropism studies in mice.

[0357] The CNS expression profile of the ARSA enzyme was assessed following ICV administration of the vector in wild-type mice. AAVhu68 was shown to predominantly transduce neurons, and experiments were performed to determine whether ARSA localized to myelin-producing oligodendrocytes. The oligodendrocyte-specific expression of ARSA supports the possibility of cross-correction of key cell types affected in MLD patients. For this study, AAVhu68.CB7.CI.hARSAcoHA.rBG was utilized, which is an AAVhu68 vector similar to AAVhu68.CB7.CI.hARSAco.rBG that encodes a human engineered ARSA enzyme tagged with a C-terminal HA peptide. Because anti-ARSA antibodies could theoretically cross-react with endogenous murine ARSA in wild-type animals, anti-hemagglutinin (HA) antibodies were used to assess ARSA expression after ICV administration. The ARSA expression profile observed following administration of this similar AAVhu68 vector is expec...

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Abstract

Provided is a recombinant adeno-associated virus (rAAV) having an AAVhu68 capsid and a vector genome which comprises a nucleic acid sequence encoding a functional human arylsulfatase A (ARSA). Also provided are a production system useful for producing the rAAV, a pharmaceutical composition comprising the rAAV, and a method of treating a subject having metachromatic leukodystrophy, or ameliorating symptoms of metachromatic leukodystrophy, or delaying progression of metachromatic leukodystrophy via administrating an effective amount of the rAAV to a subject in need thereof.

Description

Background technique [0001] Metachromatic leukodystrophy (MLD) is a monogenic autosomal recessive sphingolipidosis caused by mutations in the gene encoding the lysosomal enzyme ARSA (Von Figura et al., 2001; Gieselmann and Krageloh -Mann, 2010). ARSA deficiency results in the accumulation of its natural substrates, sulfated galactosphingolipids (galactosylceramide-3-O-sulfate and galactosylsphingosine-3-O-sulfate), Commonly known as sulfatides. In addition to Schwann cells and macrophages in the peripheral nervous system (PNS), sulfatides are also found in oligodendrocytes, microglia, and certain types of neurons in the central nervous system (CNS). Accumulates in lysosomes (Peng and Suzuki, 1987). Although the PNS and CNS are primarily affected, sulfatide storage also occurs in internal organs; most notably, the kidney, liver (Toda et al., 1990) and gallbladder (Rodriguez-Waitkus et al., 2011; McFadden and Ranganathan, 2015). [0002] MLD patients (ie, those carrying mut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/00A61K38/47A61P25/00A61P25/28A61P37/02A61P43/00
CPCA61P43/00A61P37/02A61P25/00A61P25/28C12N15/86C12Y301/06001A61K38/00C12N9/16A61K9/0085A61K47/02C12N2750/14143C12N2750/14122A61K48/005A01K2227/105A01K2227/106C12N2830/42C12N2830/50A61K48/0075A01K67/0275A01K2217/075A01K2267/0318C12N15/907C12N2750/14151C07K2319/02A61K48/00C12N2750/14121C12N2750/14171
Inventor J·霍尔多瓦J·威尔逊
Owner THE TRUSTEES OF THE UNIV OF PENNSYLVANIA