Recombinant (alpha)-L-iduronidase, methods for producing and purifying the same and methods for treating diseases caused by deficiencies thereof

An iduronidase, iduronidase technology used in molecular biology, enzymology, biochemistry and clinical medicine to solve the problems of lack of donors, infeasibility, high morbidity and mortality in MPSI

Inactive Publication Date: 2001-06-20
RES & EDUCATION INST HARBOR - UCLA MEDICAL CENT
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] No meaningful treatment for MPS I other than bone marrow transplantation
Bone marrow transplantation may be effective in treating some of the disease's symptoms, but MPS I is highly morbid and fatal and is often not feasible due to lack of suitable donors

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
  • Recombinant (alpha)-L-iduronidase, methods for producing and purifying the same and methods for treating diseases caused by deficiencies thereof
  • Recombinant (alpha)-L-iduronidase, methods for producing and purifying the same and methods for treating diseases caused by deficiencies thereof
  • Recombinant (alpha)-L-iduronidase, methods for producing and purifying the same and methods for treating diseases caused by deficiencies thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] production of recombinant iduronidase

[0077] Standard techniques such as Sambrook et al. (1987) "Molecular Cloning: A Laboratory Manual"2 nd ed, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. can be used to clone the cDNA encoding human α-L-iduronidase. The previously cloned human α-L-iduronidase cDNA was subcloned from bluescript KS as a HindIII-XbaI fragment into PRCCMV (InVitrogen). PCR amplification of bases 788-1372 (Tucker et al., Proc. Natl. Acad. Sci. USA, 78: 7684-7688 (1991)) of clone pRIR14.5 (Kakkis et al., Nucleic Acids Research, 16:7796 (1988)) An intron cassette derived from the Cot intron between exons 2 and 3 of murine immunoglobulin was augmented. The cassette included 136 bp of the 3' end of exon 2 and 5' of exon 3 The 242 bp of the end, which will remain in a correctly spliced ​​cDNA. The ATG sequence is absent in the coding region of the intron box. The intron box is cloned into the 5' HindIII position of the α-L-iduronidase cDNA Point...

Embodiment 2

[0083] Recombinant α-L-iduronidase treatment is effective

[0084] Short-term intravenous administration of purified recombinant α-L-iduronidase to 9 MPSI dogs and 6 MPSI cats has shown that the enzyme is significantly An estimated 50% or more of them are absorbed. Although the liver and spleen absorbed the greatest amount of enzyme and had the best improvement in pathology, improvement in pathology and mucopolysaccharide content was observed in many but not all tissues. Specifically, cartilage, brain and heart valves did not improve significantly. Clinical improvement was observed in one dog over 13 months of long-term treatment, but other studies were limited to 6 months or less. All dogs and most cats that received the recombinant human enzyme developed antibodies to the human product. IgG antibodies are of the complement activating type (possibly canine IgG equivalents). This phenomenon was also observed in at least 13% of Gaucher disease patients treated with glucocer...

Embodiment 3

[0088] Recombinant α-L-iduronidase therapy effective in humans

[0089] The human cDNA of α-L-iduronidase predicted a protein of 653 amino acids after cleavage of the signal peptide and a predicted molecular weight of 70,000 Daltons. Amino acid sequencing revealed the N-terminal alanine 26 giving a predicted protein of 629 amino acids. Human recombinant α-L-iduronidase has a histidine at position 8 of the mature protein. The predicted protein sequence contains 6 possible N-linked oligosaccharide modification sites. All of these sites are modified in the recombinant protein. The third and sixth positions have been shown to contain one or more mannose 6-phosphate residues that cause high affinity uptake into cells.

[0090] This peptide corresponds to amino acids 26-45 of human recombinant α-L-iduronidase, with an N-terminal alanine and the following sequence:

[0091] ala-glu-ala-pro-his-leu-val-his-val-asp-ala-ala-arg-ala-leu-trp-pro-leu-arg-arg

[0092] The recombinant e...

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
concentrationaaaaaaaaaa
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to view more

Abstract

The present invention provides a recombinant alpha -L-iduronidase and biologically active fragments and mutants thereof, methods to produce and purify this enzyme as well as methods to treat certain genetic disorders including alpha -L-iduronidase deficiency and mucopolysaccharidosis I (MPS I).

Description

field of invention [0001] The invention relates to the fields of molecular biology, enzymology, biochemistry and clinical medicine. Specifically, the present invention provides a recombinant α-L-iduronidase (α-L-iduronidase), a method for producing and purifying the enzyme and a method for treating α-L-iduronidase deficiency and mucopolysaccharide Methods for some genetic disorders of metabolic disease I (MPS I). Background of the invention [0002] Sugars play many important roles in the activities of living organisms. In addition to their metabolic role, carbohydrates are structural components in the human body that are covalently bound to many other components, such as proteins and lipids (known as glycoconjugates). For example, human connective tissue and cell membranes contain a matrix of proteins, carbohydrates and proteoglycans. The sugar moiety of the proteoglycan matrix provides important properties to the body's structure. [0003] Genetic defects in the sugar-...

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): A61K38/00A61K38/46A61K48/00A61P43/00C12N1/04C12N5/10C12N9/24C12N15/09C12N15/56C12R1/91
CPCC12Y302/01076C12N9/2402C12N9/24A61K38/00A61P43/00
Inventor E·D·卡基斯B·塔纳马奇
Owner RES & EDUCATION INST HARBOR - UCLA MEDICAL CENT
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