Pharmaceutical composition for treating cancer, containing guide RNA and endonuclease as active ingredients
Inventors
Choe, Sunghwa • Park, Mi Jin • PARK, Aiden Yeonghoon • LIM, Jung Hak • Kim, Dong Wook • IN, Sungyong • Park, Jongjin
Assignees
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Publication Number
US-11865164-B2
Publication Date
2024-01-09
Expiration Date
Abstract
A pharmaceutical composition for treating cancer, containing a crRNA and an endonuclease as active ingredients is disclosed. The composition can be customized according to the needs of patients or cell types by specifically treating cancer cells on the basis of specific binding properties of DNA and RNA. The nuclease activity of a CRISPR PLUS system, containing both an endonuclease and an exonuclease can be activated by means of the binding between crRNA and a gene specifically found in cancer cells. Therefore, the cancer treatment effect of the composition is more specific than that of other anti-cancer agents that have been developed up till now.
Core Innovation
The disclosed invention relates to anti-cancer pharmaceutical compositions and methods that treat a cancer in a subject by administering an effective amount of a composition comprising vector(s) and polynucleotides. The composition includes an isolated polynucleotide complementarily binding to a nucleic acid specifically present in cancer cells, together with isolated polynucleotides encoding an endonuclease and an exonuclease.
The method activates CRISPR-associated endonucleases with a guide RNA targeting nucleic acids specifically present in cancer cells, where activation then enables exonuclease activity that is described as sequence-activated but non-specific. The disclosed fusion concept provides endonuclease and exonuclease as a fusion protein, combining an endonuclease selected from a group of CRISPR-associated proteins with an exonuclease selected from a specified list.
The nucleic acid specifically present in cancer cells is selected from genes for which copy number variation (CNV) is at least 4, and dependent refinements increase the CNV threshold to at least 7 and broaden target types to SNP, CNV, SV, insertion, and deletion. The disclosure frames a multi-targeting capability in connection with high-CNV regions and describes exemplified target genes including p53, PTEN, APC, MSH2, HBV/HCV, and EGFR.
Claims Coverage
The independent claim covers a cancer treatment method using a CRISPR-associated endonuclease and an exonuclease packaged as endonuclease and exonuclease encoding polynucleotides, including a complementarily binding nucleic-acid targeting polynucleotide, with the endonuclease and exonuclease being a fusion protein and targeting nucleic acids specifically present in cancer cells selected by a CNV threshold. The inventive features are organized across 5 dependent claims that refine the nucleic-acid type, structural subtypes, CNV threshold, example mutant gene targets, and allowable administration routes.
Cancer treatment using vector with cancer-specific complementarily binding polynucleotide and endonuclease/exonuclease-encoding polynucleotides
A method of treating a cancer in a subject comprising administering an effective amount of a composition comprising a vector containing an isolated polynucleotide complementarily binding to a nucleic acid specifically present in cancer cells, an isolated polynucleotide encoding an endonuclease, and an isolated polynucleotide encoding an exonuclease.
Exonuclease selection from defined list
The exonuclease is selected from the group consisting of RecBCD, RecE, RecJ, T5, Exo I, Exo III, Exo VII, Lexo, TREX2, Exoribonuclease T, TREXI, Mungbean exonuclease, Lambda, and a combination thereof.
Cancer-specific nucleic acid selected by CNV threshold (at least 4)
The nucleic acid specifically present in cancer cells is selected from the genes of which copy number variation (CNV) is at least 4.
CRISPR-associated endonuclease selected from defined Cas/Csf/Csy groups
The endonuclease is a clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein selected from a group consisting of Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9, Cas10, Cas12a, Cas12b, Cas12c, Cas12d, Cas12e, Cas12g, Cas12h, Cas12i, Cas13a, Cas13b, Cas13c, Cas13d, Cas14, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, CsMT2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, and Csf4.
Fusion protein of endonuclease and exonuclease
The endonuclease and the exonuclease are of a fusion protein.
Across the independent claim and dependent refinements, coverage centers on a CRISPR-associated endonuclease plus an exonuclease provided via a fusion protein, together with a vector carrying an isolated polynucleotide that complementarily binds to a cancer-specific nucleic acid. The cancer specificity is defined by CNV thresholds (at least 4, and at least 7 in a dependent claim) and further specified in dependent claims by SNP/SV/insertion/deletion types and structural subtypes, along with example mutant gene targets and specified administration routes.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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