WO1991018624A1 - Ribozyme mediated reversal of transformation by cleavage of the hras oncogene rna - Google Patents

Ribozyme mediated reversal of transformation by cleavage of the hras oncogene rna Download PDF

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WO1991018624A1
WO1991018624A1 PCT/US1990/006226 US9006226W WO9118624A1 WO 1991018624 A1 WO1991018624 A1 WO 1991018624A1 US 9006226 W US9006226 W US 9006226W WO 9118624 A1 WO9118624 A1 WO 9118624A1
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ribozyme
plasmid
hras
rna
cleavage
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PCT/US1990/006226
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Kevin J. Scanlon
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City Of Hope
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Priority to PCT/US1990/007459 priority Critical patent/WO1991018625A1/en
Priority to AU71521/91A priority patent/AU7152191A/en
Publication of WO1991018624A1 publication Critical patent/WO1991018624A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1135—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against oncogenes or tumor suppressor genes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/82—Translation products from oncogenes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/12—Type of nucleic acid catalytic nucleic acids, e.g. ribozymes
    • C12N2310/121—Hammerhead

Definitions

  • This invention relates to a ribozyme effective to cleave oncogene RNA. More particularly the invention relates to a ribozyme effective to cleave Hras RNA at a GUC site on codon 12 where an activating point mutation may appear, to a plasmid into which said ribozyme has been cloned, to mammalian cells into which said plasmid has been transfected and which express said ribozyme and to cancer therapy which entails the administration of such ribozymes per se or of such plasmids to induce in vivo expression of such ribozymes.
  • These ribozymes or ribozyme plasmids can reverse the transformation process to convert malignant cells back to the normal cell phenotype.
  • Ribozyme cleavage of various RNA transcripts is known. See, e.g., published PCT applications no. 89/05852 and no. 88/04300 and pending U.S. Application 9/401,613 filed August 31, 1989.
  • Hras oncogene activates the Hras oncogene to yield a gene product which malignantly transforms cells and may cause a spectrum of neoplasms including human bladder carcinoma.
  • no known cancer therapy protocol implicates this phenomenon.
  • Su mary of the Invention Normal Hras gene RNA does not include a ribozyme cleavage site on codon 12.
  • the point mutation which yields a malignancy inducing Hras transcript does include such a cleavage site.
  • This invention provides a ribozyme effective to cleave the mutated gene both in vitro and in vivo, but not the normal gene.
  • the ribozyme of the invention may be administered by a known delivery system such as a liposome or by other means known to the art. It may also be administered in the form of a vector (i.e., RNA tumor virus) into which it has been cloned and which will express the ribozyme.
  • a vector i.e., RNA tumor virus
  • the invention also includes a ribozyme which inhibits C-fos expression in response to cis-platin or other stimuli.
  • the Hras RNA included in mutated gene transcript includes a GUC site appropriate for ribozyme cleavage as shown in II. II. 5' GUG GUG GGC GCC GUC GGU GUG GGC AAG 3' +8 9 10 11 12 13 14 15 16 A ribozyme effective to catalytically cleave the mutated Hras DNA at the GUC cleavage site in functional association therewith is shown in III.
  • nucleotide which is implicated in the mutation is shown by an open circle. As indicated, the nucleotide is "G” in the normal sequence and "U” in the mutation.
  • the ribozyme per se may be synthesized in known manner by use of a commercially available synthesizer produced, for example, by Applied Biosystems, Inc. or Milligen.
  • a double stranded DNA molecule having one strand which upon transformation yields the desired ribozyme, is synthesized from single stranded oligodeoxynucleotide molecules.
  • This molecule is then cloned into a plasmid capable of synthesizing the ribozyme in vivo when transfected into a mammalian cell.
  • Appropriate promoters, e.g., T7 RNA polymerase, and terminal sequences may be present and follow the "ribozyme" component of the DNA insert to be cloned into the plasmid.
  • nucleotide sequences having sufficient overlapping base pairs are amplified by the polymerase chain reaction to provide the insert to be cloned.
  • the 3' and 5' termini include restriction sites to insure the correct positioning of the insert in the plasmid.
  • a T 7 RNA polymerase promoter may be positioned at the 5' terminus of the "ribozyme" to accommodate in vitro cleavage.
  • V.A and V.B One appropriate double stranded construct is shown by V.A and V.B.
  • the single stranded synthetic oligodeoxynucleotides with flanking Sal I and Hind III restriction sites are annealed to provide an insert with sticky ends including the ribozyme to be cloned into the plasmid.
  • T7 RNA polymerase promoter sequence shown in V.A may be omitted if _in vitro cleavage of Hras RNA is not contemplated.
  • a preferred plasmid is pH-0-Apr.l, more specifically _pH- ⁇ Apr.l-neo which yields the corresponding vector pH- / 3 Apr.1-Hras-R in which R is the double stranded PCR product, one strand of which, V.A includes the Hras ribozyme.
  • Other plasmids useful in the invention include pH pKoneo, the pSV2 cat plasmid and pMAMneo.
  • the DNA fragments provided by the PCR amplification product are cloned by sticky end ligation into a selected site of the plasmid.
  • FIGURES Figure 1 is a block diagram schematic illustration of certain steps included in the invention.
  • Figure 2 illustrates a plasmid including a ribozyme of the type shown, in III and IV.
  • the sequence below the plasmid illustrates the "ras-ribozyme" insert.
  • the Hras ribozymes or the plasmid vectors which express such ribozymes of the invention may be administered by injection of appropriate delivery systems such as lipomes.
  • one aspect of the invention includes liposomes in which the Hras ribozymes are encapsulated or are included in the lyposomal bilayers.
  • EXAMPLE 1 A control group and an experimental group of nude mice were treated with one of the following treatments:
  • the control group received the EJ cells without the H-ras ribozyme. Seven of eight control mice died with an average survival of 4.5 weeks. Pathology studies revealed metastatic invasion of lung, kidney and liver of seven out of eight control mice within four to six weeks of treatment.
  • the experimental group received the EJ cells with the H-ras ribozyme. This group had only three deaths after 11.0 weeks. The pathologoy of these dead animals indicated no invasive tumor. This data is summarized in Table 1.

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  • Life Sciences & Earth Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract

Ribozyme mediated transformation of malignant cancer cells to normal cancer cells is described.

Description

RIBOZYME MEDIATED REVERSAL OF TRANSFORMATION BY CLEAVAGE OF THE HRAS ONCOGENE RNA
Field of the Invention
This application is a continuation-in-part of application PCT/US/90/03218, filed 07 June, 1990, which is incorporated herein by reference.
This invention relates to a ribozyme effective to cleave oncogene RNA. More particularly the invention relates to a ribozyme effective to cleave Hras RNA at a GUC site on codon 12 where an activating point mutation may appear, to a plasmid into which said ribozyme has been cloned, to mammalian cells into which said plasmid has been transfected and which express said ribozyme and to cancer therapy which entails the administration of such ribozymes per se or of such plasmids to induce in vivo expression of such ribozymes. These ribozymes or ribozyme plasmids can reverse the transformation process to convert malignant cells back to the normal cell phenotype.
Background of the Invention Ribozyme cleavage of various RNA transcripts is known. See, e.g., published PCT applications no. 89/05852 and no. 88/04300 and pending U.S. Application 9/401,613 filed August 31, 1989.
It is known that a point mutation on codon 12 activates the Hras oncogene to yield a gene product which malignantly transforms cells and may cause a spectrum of neoplasms including human bladder carcinoma. To date no known cancer therapy protocol implicates this phenomenon. Su mary of the Invention Normal Hras gene RNA does not include a ribozyme cleavage site on codon 12. The point mutation which yields a malignancy inducing Hras transcript does include such a cleavage site. This invention provides a ribozyme effective to cleave the mutated gene both in vitro and in vivo, but not the normal gene. It accordingly provides unique therapy for bladder carcinoma and other malignancies which may be induced by cells transformed by the mutated Hras gene product. The ribozyme of the invention may be administered by a known delivery system such as a liposome or by other means known to the art. It may also be administered in the form of a vector (i.e., RNA tumor virus) into which it has been cloned and which will express the ribozyme.
The invention also includes a ribozyme which inhibits C-fos expression in response to cis-platin or other stimuli.
Detailed Description of the Invention The DNA sequence flanking codon 12 of the human Hras gene is illustrated by I:
[T] I. 5' GTG GTG GGC GCC GGC CGT GTG GGC AAG 3' 8 9 10 11 12 13 14 15 16
The mutation which activates the Hras gene is shown in brackets above position 2 of codon 12.
The Hras RNA included in mutated gene transcript according includes a GUC site appropriate for ribozyme cleavage as shown in II. II. 5' GUG GUG GGC GCC GUC GGU GUG GGC AAG 3' +8 9 10 11 12 13 14 15 16 A ribozyme effective to catalytically cleave the mutated Hras DNA at the GUC cleavage site in functional association therewith is shown in III.
III. Hras RNA
5'- G GGC GCC GUC GGU GUG GGC - 3 C CCG CGG CA CCA CAC CCG 5' 3' A Cu
GA A U Ribozyme G AG
C-G A-U G-C G-C
G A GU
Another representation of the ribozyme is shown in IV.
Figure imgf000006_0001
Figure imgf000006_0004
Figure imgf000006_0002
<CGGUGUGGGC-3' cHras RNA catalytic strand
Figure imgf000006_0003
In IV, the nucleotide which is implicated in the mutation is shown by an open circle. As indicated, the nucleotide is "G" in the normal sequence and "U" in the mutation.
The ribozyme per se may be synthesized in known manner by use of a commercially available synthesizer produced, for example, by Applied Biosystems, Inc. or Milligen.
In the preferred practice of the invention a double stranded DNA molecule having one strand, which upon transformation yields the desired ribozyme, is synthesized from single stranded oligodeoxynucleotide molecules. This molecule is then cloned into a plasmid capable of synthesizing the ribozyme in vivo when transfected into a mammalian cell. Appropriate promoters, e.g., T7 RNA polymerase, and terminal sequences may be present and follow the "ribozyme" component of the DNA insert to be cloned into the plasmid. Preferably appropriate nucleotide sequences having sufficient overlapping base pairs are amplified by the polymerase chain reaction to provide the insert to be cloned. The 3' and 5' termini include restriction sites to insure the correct positioning of the insert in the plasmid. A T7 RNA polymerase promoter may be positioned at the 5' terminus of the "ribozyme" to accommodate in vitro cleavage. One appropriate double stranded construct is shown by V.A and V.B.
V.A Sequence 1
T7 RNA polymerase promoter 5' GGT CGA CTA ATA CGA CTC ACT ATA GGC CCA CAC CCT GAT Sal I 3' C GTG GGA CTA
V.B Sequence 2
GA - 3'
CTC AGG CAC TCC TGC TTT GCC GCG GGT TCG AAC - 5'
Hind III
The single stranded synthetic oligodeoxynucleotides with flanking Sal I and Hind III restriction sites are annealed to provide an insert with sticky ends including the ribozyme to be cloned into the plasmid.
The T7 RNA polymerase promoter sequence shown in V.A may be omitted if _in vitro cleavage of Hras RNA is not contemplated.
The sal I and Hind III sequences are included to insure that the double stranded product which results from PCR amplification of the V.A-V.B construct appears in the proper orientation when cloned into a plasmid to produce a vector. The selection of an appropriate plasmid is within the skill of the art. A preferred plasmid is pH-0-Apr.l, more specifically _pH-β Apr.l-neo which yields the corresponding vector pH-/3 Apr.1-Hras-R in which R is the double stranded PCR product, one strand of which, V.A includes the Hras ribozyme. Other plasmids useful in the invention include pH pKoneo, the pSV2 cat plasmid and pMAMneo.
The DNA fragments provided by the PCR amplification product are cloned by sticky end ligation into a selected site of the plasmid.
Cells transfected, e.g., by lipofection or electroporation, with the vector pH-ø-Apr.l Hras R express the Hras ribozyme shown in III and IV.
DESCRIPTION OF THE FIGURES Figure 1 is a block diagram schematic illustration of certain steps included in the invention.
Figure 2 illustrates a plasmid including a ribozyme of the type shown, in III and IV. The sequence below the plasmid illustrates the "ras-ribozyme" insert.
The Hras ribozymes or the plasmid vectors which express such ribozymes of the invention may be administered by injection of appropriate delivery systems such as lipomes. Hence, one aspect of the invention includes liposomes in which the Hras ribozymes are encapsulated or are included in the lyposomal bilayers. In vivo ribozymes expressed by cells transfected with a plasmid, as described, express the ribozyme shown in III or IV which cleaves the RNA expressed by malignant Hras genes as evidenced by Example 1. EXAMPLE 1 A control group and an experimental group of nude mice were treated with one of the following treatments:
1. EJ human bladder carcinoma cells exposed (treated) to the plasmid pϋ-β Apr.l-neo (no ribozyme) ; and
2. EJ human bladder carcinoma cells exposed (treated) to the Figure 2 plasmid pH-β Apr.l-neo containing the H-ras ribozyme as shown by Figure 2.
Cells (2 x 106) from each group were injected into the bladder of the nude mice.
The control group received the EJ cells without the H-ras ribozyme. Seven of eight control mice died with an average survival of 4.5 weeks. Pathology studies revealed metastatic invasion of lung, kidney and liver of seven out of eight control mice within four to six weeks of treatment.
The experimental group received the EJ cells with the H-ras ribozyme. This group had only three deaths after 11.0 weeks. The pathologoy of these dead animals indicated no invasive tumor. This data is summarized in Table 1.
TABLE 1 Mice Control EJ pH ffHrasR
Dead/Mice 7/8 3/53
% Survival 12% 94.4% In summary, the control animals had a 12% survival rate at six weeks while 94.4% of the experimental animals (with H-ras ribozyme) were alive after 11.0 weeks.

Claims

1. A ribozyme which cleaves Hras RNA at a GUC cleavage site.
2. A ribozyme having the sequence of the ribozyme shown in III.
3. A plasmid having as a cloned insert, a double stranded DNA fragment one strand of said fragment having the sequence of the ribozyme shown in III.
4. A cell including a plasmid, as defined by
Claim 3, said cell expressing a ribozyme as shown in III.
5. The plasmid depicted by Figure 2.
6. A method of converting a malignant cancer cell to a nonmalignant cell which comprises incorporating a plasmid, as defined by Claim 5, into said cell.
7. A method as defined by claim 6 in which said malignant cancer cell is a malignant EJ human bladder cancer cell.
8. A method for treating human bladder carcinoma which comprises administering to a patient in need of such treatment an effective amount of a plasmid, as defined by Claim 5.
PCT/US1990/006226 1990-06-07 1990-11-01 Ribozyme mediated reversal of transformation by cleavage of the hras oncogene rna WO1991018624A1 (en)

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USPCT/US90/03218 1990-06-07

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WO1993023057A1 (en) * 1992-05-14 1993-11-25 Ribozyme Pharmaceuticals, Inc. Method and reagent for inhibiting cancer development
US5496698A (en) * 1992-08-26 1996-03-05 Ribozyme Pharmaceuticals, Inc. Method of isolating ribozyme targets
US5599704A (en) * 1992-08-26 1997-02-04 Ribozyme Pharmaceuticals, Inc. ErbB2/neu targeted ribozymes
US5610052A (en) * 1992-08-26 1997-03-11 Ribozyme Pharmaceuticals Inc. Enzymatic RNA with activity to ras
US5612215A (en) * 1992-12-07 1997-03-18 Ribozyme Pharmaceuticals, Inc. Stromelysin targeted ribozymes
US5616488A (en) * 1992-12-07 1997-04-01 Ribozyme Pharmaceuticals, Inc. IL-5 targeted ribozymes
US5616490A (en) * 1992-12-07 1997-04-01 Ribozyme Pharmaceuticals, Inc. Ribozymes targeted to TNF-α RNA
US5639655A (en) * 1993-01-19 1997-06-17 Ribozyme Pharmaceuticals, Inc. PML-RARA targeted ribozymes
US5646042A (en) * 1992-08-26 1997-07-08 Ribozyme Pharmaceuticals, Inc. C-myb targeted ribozymes
US5658780A (en) * 1992-12-07 1997-08-19 Ribozyme Pharmaceuticals, Inc. Rel a targeted ribozymes
US5750390A (en) * 1992-08-26 1998-05-12 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of diseases caused by expression of the bcl-2 gene
US5811300A (en) * 1992-12-07 1998-09-22 Ribozyme Pharmaceuticals, Inc. TNF-α ribozymes
US5989906A (en) * 1992-05-14 1999-11-23 Ribozyme Pharmaceuticals, Inc. Method and reagent for inhibiting P-glycoprotein (mdr-1-gene)
US6080851A (en) * 1992-12-04 2000-06-27 American Home Products Corporation Ribozymes with linked anchor sequences
US6103890A (en) * 1994-05-18 2000-08-15 Ribozyme Pharmaceuticals, Inc. Enzymatic nucleic acids that cleave C-fos
US6132967A (en) * 1992-12-07 2000-10-17 Ribozyme Pharmaceuticals, Inc. Ribozyme treatment of diseases or conditions related to levels of intercellular adhesion molecule-1 (ICAM-1)
US6492512B1 (en) 1992-08-26 2002-12-10 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of lung cancer and other malignancies caused by the deregulation of L-MYC gene expression
US6544755B1 (en) 1992-08-26 2003-04-08 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of diseases by expression of the c-Myc gene
US6656731B1 (en) 1997-09-22 2003-12-02 Max Planck Gesellschaft Zur Forderung Der Wissenschaften E.V. Nucleic acid catalysts with endonuclease activity
WO2024026474A1 (en) 2022-07-29 2024-02-01 Regeneron Pharmaceuticals, Inc. Compositions and methods for transferrin receptor (tfr)-mediated delivery to the brain and muscle
WO2024098002A1 (en) 2022-11-04 2024-05-10 Regeneron Pharmaceuticals, Inc. Calcium voltage-gated channel auxiliary subunit gamma 1 (cacng1) binding proteins and cacng1-mediated delivery to skeletal muscle
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Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5246921A (en) * 1990-06-26 1993-09-21 The Wistar Institute Of Anatomy And Biology Method for treating leukemias
US6469158B1 (en) * 1992-05-14 2002-10-22 Ribozyme Pharmaceuticals, Incorporated Synthesis, deprotection, analysis and purification of RNA and ribozymes
US5804683A (en) * 1992-05-14 1998-09-08 Ribozyme Pharmaceuticals, Inc. Deprotection of RNA with alkylamine
US5686599A (en) * 1992-05-14 1997-11-11 Ribozyme Pharmaceuticals, Inc. Synthesis, deprotection, analysis and purification of RNA and ribozymes
US5714383A (en) * 1992-05-14 1998-02-03 Ribozyme Pharmaceuticals, Inc. Method and reagent for treating chronic myelogenous leukemia
US5977343A (en) 1992-05-14 1999-11-02 Ribozyme Pharmaceuticals, Inc. Synthesis, deprotection, analysis and purification of RNA and ribozymes
PT808898E (en) * 1996-05-24 2004-10-29 Aventis Pharma Gmbh REAGENT AND METHOD FOR INHIBITING N-RAS EXPRESSION

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0321201A2 (en) * 1987-12-15 1989-06-21 Gene Shears Pty. Limited Ribozymes

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4987071A (en) * 1986-12-03 1991-01-22 University Patents, Inc. RNA ribozyme polymerases, dephosphorylases, restriction endoribonucleases and methods

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0321201A2 (en) * 1987-12-15 1989-06-21 Gene Shears Pty. Limited Ribozymes

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Volume 112, No. 13, issued 26 March 1990, CAMERON et al., "Specific Gene Suppression by Engineered Ribozymes in Monkey Cells", see page 232, Column 1, the Abstract No. 1135276; & PROC. NATL. ACAD. SCI. USA. 1989, 86(23), 9139-43, (Eng.). *
CHEMICAL ABSTRACTS, Volume 112, No. 19, issued 07 May 1990, SARVER et al., "Ribozymes as Potential and HIV-1 Therapeutic Agents", see page 420, Column 1, the Abstract No. 175480q; & SCIENCE, 1990, 247(4947), 1225-5, (Eng.). *
CHEMICAL ABSTRACTS, Volume 112, No. 7, issued 12 February 1990, GELACH et al., "Synthetic Ribozymes for In Vivo Inactivation of Prokaryotic or Eakortyotic RNA Transcripts", see pages 336-337, Column 2, and 1, the Abstract No. 51284; & EP,A,321 201, 21 June 1989. *
NATURE, Volume 344, issued 05 April 1990, J. PERREAULT et al., "Mixed Deoxyribo-and Ribouligo-Nucleotides with Catalytic Activity", see pages 565-567. *
PROC. NATL. ACAD. SCI. USA, Volume 86, issued December 1989, CAMERON et al., "Specific Gene Suppression by Engineered Ribozymes in Monkey Cells", see pages 9139-9143. *

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WO1993023057A1 (en) * 1992-05-14 1993-11-25 Ribozyme Pharmaceuticals, Inc. Method and reagent for inhibiting cancer development
US5989906A (en) * 1992-05-14 1999-11-23 Ribozyme Pharmaceuticals, Inc. Method and reagent for inhibiting P-glycoprotein (mdr-1-gene)
US5801158A (en) * 1992-08-26 1998-09-01 Ribozyme Pharmaceuticals, Inc. Enzymatic RNA with activity to RAS
US5496698A (en) * 1992-08-26 1996-03-05 Ribozyme Pharmaceuticals, Inc. Method of isolating ribozyme targets
US5599704A (en) * 1992-08-26 1997-02-04 Ribozyme Pharmaceuticals, Inc. ErbB2/neu targeted ribozymes
US5610052A (en) * 1992-08-26 1997-03-11 Ribozyme Pharmaceuticals Inc. Enzymatic RNA with activity to ras
US6544755B1 (en) 1992-08-26 2003-04-08 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of diseases by expression of the c-Myc gene
US6492512B1 (en) 1992-08-26 2002-12-10 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of lung cancer and other malignancies caused by the deregulation of L-MYC gene expression
US5646042A (en) * 1992-08-26 1997-07-08 Ribozyme Pharmaceuticals, Inc. C-myb targeted ribozymes
US5817796A (en) * 1992-08-26 1998-10-06 Stinchcomb; Dan T. C-myb ribozymes having 2'-5'-linked adenylate residues
US5750390A (en) * 1992-08-26 1998-05-12 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of diseases caused by expression of the bcl-2 gene
US6080851A (en) * 1992-12-04 2000-06-27 American Home Products Corporation Ribozymes with linked anchor sequences
US5616490A (en) * 1992-12-07 1997-04-01 Ribozyme Pharmaceuticals, Inc. Ribozymes targeted to TNF-α RNA
US5612215A (en) * 1992-12-07 1997-03-18 Ribozyme Pharmaceuticals, Inc. Stromelysin targeted ribozymes
US5731295A (en) * 1992-12-07 1998-03-24 Ribozyme Pharmaceuticals, Inc. Method of reducing stromelysin RNA via ribozymes
US5658780A (en) * 1992-12-07 1997-08-19 Ribozyme Pharmaceuticals, Inc. Rel a targeted ribozymes
US5811300A (en) * 1992-12-07 1998-09-22 Ribozyme Pharmaceuticals, Inc. TNF-α ribozymes
US5616488A (en) * 1992-12-07 1997-04-01 Ribozyme Pharmaceuticals, Inc. IL-5 targeted ribozymes
US6132967A (en) * 1992-12-07 2000-10-17 Ribozyme Pharmaceuticals, Inc. Ribozyme treatment of diseases or conditions related to levels of intercellular adhesion molecule-1 (ICAM-1)
US6410224B1 (en) 1992-12-07 2002-06-25 Ribozyme Pharmaceuticals, Inc. Ribozyme treatment of diseases or conditions related to levels of NF-κB
US6436644B1 (en) 1992-12-07 2002-08-20 Ribozyme Pharmaceuticals, Inc. Ribozyme treatment of diseases or conditions related to levels of intercellular adhesion molecule-1 (CAM-1)
US5639655A (en) * 1993-01-19 1997-06-17 Ribozyme Pharmaceuticals, Inc. PML-RARA targeted ribozymes
US6103890A (en) * 1994-05-18 2000-08-15 Ribozyme Pharmaceuticals, Inc. Enzymatic nucleic acids that cleave C-fos
US6656731B1 (en) 1997-09-22 2003-12-02 Max Planck Gesellschaft Zur Forderung Der Wissenschaften E.V. Nucleic acid catalysts with endonuclease activity
WO2024026474A1 (en) 2022-07-29 2024-02-01 Regeneron Pharmaceuticals, Inc. Compositions and methods for transferrin receptor (tfr)-mediated delivery to the brain and muscle
WO2024098002A1 (en) 2022-11-04 2024-05-10 Regeneron Pharmaceuticals, Inc. Calcium voltage-gated channel auxiliary subunit gamma 1 (cacng1) binding proteins and cacng1-mediated delivery to skeletal muscle
WO2024107765A2 (en) 2022-11-14 2024-05-23 Regeneron Pharmaceuticals, Inc. Compositions and methods for fibroblast growth factor receptor 3-mediated delivery to astrocytes

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