TW201201819A - Treatment of BCL2-like 11 (BCL2L11) related diseases by inhibition of natural antisense transcript to BCL2L11 - Google Patents

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of BCL2-like 11 (BCL2L11), in particular, by targeting natural antisense polynucleotides of BCL2-like 11 (BCL2L11). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of BCL2L11.

Description

201201819 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION Embodiments of the invention include oligonucleotides and related molecules that modulate the performance and/or function of BCL2L11. The present application claims priority to U.S. Provisional Patent Application Serial No. 61/346,252, filed on Jan. [Prior Art] DNA-RNA and RNA-RNA hybridization are important for many nucleic acid functional aspects including DNA replication, transcription, and translation. Hybridization is also critical for the detection of specific nucleic acids or the various techniques that alter their performance. For example, antisense nucleotides disrupt gene expression by hybridizing to a target RNA, thereby interfering with RNA splicing, transcription, translation, and replication. Antisense DNA has the additional property that DNA-RNA hybrids can be used as receptors for ribonuclease digestion, which is present in most cell types. The antisense molecule can be delivered to a cell, such as in the case of an oligodeoxynucleotide (ODN), or it can be expressed as an RNA molecule from an endogenous gene. The FDA recently approved the antisense drug VITRAVENETM (for the treatment of cytomegalovirus retinitis), indicating that the antisense drug has therapeutic utility. BRIEF DESCRIPTION OF THE DRAWINGS The Summary is provided to introduce a summary of the invention This Summary is submitted with the understanding that it is not to be construed as limiting or limiting the scope or meaning of the scope of the application. In one embodiment, the invention provides a method of inhibiting the action of a natural antisense transcript 156341.doc 201201819 by up-regulating an antisense oligonucleotide by using a shoe to any region of a natural antisense transcript The righteous gene is reached. It is also contemplated herein to inhibit natural antisense transcripts by siRNA, ribozymes and small molecules, which are considered to be within the scope of the invention. One embodiment provides a method of modulating the function and/or expression of a BCL2L11 polynucleotide in a patient's cells or tissues in vivo or in vitro, comprising subjecting the cells or tissues to a length of 5 to 30 nucleotides The oligonucleotide is contacted, wherein the oligonucleotide has at least 50% sequence identity to the reverse complement of the polynucleotide, the polynucleotide comprising nucleotides 1 to 3026 of SEQ ID NO: 2 and 5 to 30 contiguous nucleotides within nucleotides 1 to 1512 of SEQ ID NO: 3 ' thereby modulating the function and/or expression of BCL2L11 polynucleotides in a patient's cells or tissues in vivo or in vitro. In one embodiment, the oligonucleotide targets a natural antisense sequence of a BCL2L11 polynucleotide (eg, the nucleotides set forth in SEQ ID NOS: 2 and 3), and any variants thereof, allelic Examples of genes, homologs, mutants, derivatives, fragments and complementary sequences of antisense oligonucleotides are set forth in SEQ ID NOs: 4 to 13 . Another embodiment provides for the regulation of patient cells in vivo or in vitro. Or a method of functioning and/or expressing a BCL2L11 polynucleotide in a tissue comprising contacting the cells or tissues with an antisense oligonucleotide of 5 to 30 nucleotides in length, wherein the oligonucleotide The reverse complement of the BCL2L11 polynucleotide antisense transcript has at least 50% sequence identity; thereby modulating the function and/or expression of the BCL2L11 polynucleotide in a patient cell or tissue in vivo or in vitro. 156341.doc 201201819 Another embodiment provides a method of modulating the function and/or expression of a BCL2L11 polynucleotide in a cell or tissue of a patient in vivo or in vitro, comprising culturing a cell or tissue such as shai with a length of 5 to 30 One nucleotide antisense oligonucleotide contacts 'where the nucleotide has at least 50% sequence identity with the antisense oligonucleotide of the BCL2L11 antisense polynucleotide; thus in vivo or in vivo Externally regulates the function and/or performance of BCL2L11 polynucleotides in a patient's cells or tissues. In one embodiment, the composition comprises one or more antisense oligonucleotides that bind to a sense and/or antisense BCL2L11 polynucleotide. In one embodiment, the oligonucleotide comprises one or more modified or substituted nucleotides. In one embodiment the 'oligonucleotide comprises one or more modified linkages. In yet another embodiment, the modified nucleotide comprises a modified base comprising the following: a phosphorothioate, a phosphonium phosphonate, a peptide nucleic acid, a 2, a fluorenyl group, a fluoro- or a carbon, a methylene group or other lock. Nucleic acid (LNA) molecule. Preferably, the modified nucleotide acid-locked nucleic acid molecule comprises a-L-LNA. In one embodiment, the oligonucleotide is administered to the patient subcutaneously, intramuscularly, intravenously or intraperitoneally. In an embodiment, the oligo(tetra) acid is administered as a pharmaceutical composition. The treatment regimen includes administering the antisense compound to the patient at least once; however, the treatment can be modified to include multiple doses over a set period of time. The treatment can be combined with - or a plurality of other types of therapies. 5 In the examples, the oligonucleotide is encapsulated in a liposome or an accessory molecule (e.g., cholesterol, TAT peptide). 156341.doc 201201819 Other aspects are set out below. [Embodiment] Several aspects of the present invention are explained below by way of example application with reference to the explanation. It is to be understood that the specific details, aspects However, it will be readily apparent to those skilled in the art that the present invention may be practiced without the use of one or more specific details or other methods. The present invention is not limited to the order of the various acts or events, as some of the acts may occur in different orders and/or concurrently with other acts or events. In addition, not all illustrated acts or events may be required to implement the methods of the invention. All of the genes, gene names, and gene products disclosed herein are intended to correspond to homologs from any species to which the compositions and methods disclosed herein can be applied. Thus, such terms include, but are not limited to, human and mouse genes and gene products. It is to be understood that the disclosure is intended to be illustrative only, and is not intended to be limiting, unless the context clearly indicates otherwise. Thus, for example, in certain embodiments relating to mammalian nucleic acid and amino acid sequences, the genes disclosed herein are intended to encompass homologous and/or orthologous genes and gene products from other animals' such other animals. Includes, but is not limited to, other mammals, fish, amphibians, reptiles, and birds. In one embodiment, the gene or nucleic acid sequence is human. The terminology used herein is for the purpose of the description of the embodiments and As used herein, the singular forms "a", "a", "the", "the", "the", "the", "the", "the", and "the" are also intended to include the plural. In addition, the terms "ineluding", "ineludes", /, (1^7丨11§), "has", "have" are used in the detailed description and/or patent application. The terms "with", or variations thereof, are intended to mean a range of inclusions in a manner similar to the term "comprising." The term "about" or "approximately acceptable range of acceptable values for a particular value" is intended to be determined by those skilled in the art, and is determined in part by the manner in which the value is measured or measured, that is, the limitations of the measurement system. . For example, according to industry practice, "about" can mean within one or more standard deviations. Alternatively, "about" can mean a range that differs from a given value by at most 2%, preferably at most 10%, more preferably at most 5%, and even more preferably at most 1%. Alternatively, especially for biological systems or processes, the term may mean within an order of magnitude, preferably within 5 times, and more preferably within 2 times of a numerical value. In the case of a specific value stated in the scope of the application and the scope of the patent application, unless otherwise stated, the term "approximately" means that it is within the acceptable tolerance of the specified value. The term "mRNA" as used herein refers to a currently known mRNA transcript of a targeted gene, and any other transcript that can be elucidated. An "antisense oligonucleotide" or "antisense compound" means an RNA or DNA molecule that binds to another RNA or DNA (target RNA, DNA). For example, if an RNA oligonucleotide is used, it binds to another RNA by RNA-RNA interaction and changes the activity of the dry RNA. Antisense oligonucleotides can up- or down-regulate the performance and/or function of a particular polynucleic acid. This definition actually contains I56341.doc 201201819 containing any foreign RNA or DNA molecule that can be used for therapeutic, diagnostic, or other aspects. Such molecules include, for example, antisense RNA or DNA molecules, interfering RNA (RNAi), microRNAs, declining RNA molecules, siRNA, enzyme RNA, therapeutically editable RNA and RNA agonists and antagonists, antisense oligomeric compounds An antisense oligonucleotide, an external leader sequence (EGS) oligonucleotide, an alternative splicing, a primer, a probe, and other oligomeric compounds that hybridize to at least a portion of the target nucleic acid. Thus, such compounds can be introduced as single-stranded, double-stranded, partially single-stranded, or cyclic oligomeric compounds. In the context of the present invention, the term "oligonucleotide" refers to an oligo or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or a mimetic thereof. The term "oligonucleotide" also encompasses natural and/or modified monomers or linked linear or cyclic oligomers, including deoxyribonucleosides, ribonucleosides, substituted and alpha-rotated Configuration form, peptide nucleic acid (PNA), locked nucleic acid (LNA), thio acid complex, methyl phosphonate, and the like. Oligonucleotides can specifically bind to a target polynucleotide such as Watson-Crick type base pairing, Ho0gsteen type base pairing And so on. Oligonucleotides can be "chimeric" oligonucleotides, i.e., composed of different regions. In the context of the present invention, a "chimeric" compound is an oligonucleotide comprising two or more chemical regions, such as a DNA region, an RNA region, a PNA region, and the like. In the case of an oligonucleotide compound, each chemical region consists of at least one monomer unit (i.e., nucleotide). Such oligonucleotides typically include at least one region that modifies the oligonucleotide to reveal one or more desired properties. Desirable properties of the oligonucleotide include, but are not limited to, 156341.doc 201201819 (for example): increasing resistance to nuclease degradation, increasing cellular uptake, and/or increasing binding affinity for a target nucleic acid. Different regions of the oligonucleotide may thus have different properties. The chimeric oligonucleotides of the invention can be formed into a hybrid structure of two or more of the above oligonucleotides, modified nucleotides, oligonucleosides and/or nucleus nucleotide analogs. Oligonucleotides can be composed of regions that can be joined in a "register", i.e., a single system of continuous linkages (e. g., in native DNA), or via a spacer. The spacers are intended to form a covalent "bridge" between the regions and, in the preferred case, have a length of no more than about 100 carbon atoms. The spacers can have different functions, such as having a positive or negative charge, having specific nucleic acid binding properties (chimeric agents, groove binders, toxins, fluorophores, etc.), having lipophilic properties, and inducing specific secondary structures (eg, , to induce the acridine-containing peptide). As used herein, "BCL2L11" and "BCL2U-like" include all family members, mutants, alleles, fragments @, species, coding and non-coding sequences, sense and antisense polynucleotide chains, and the like. The words BCL2 U, BCL2LU, BAM, cell death Bcl2 interacting mediator, Bcl2_L_u, face, ugly-^, face_β6, ΒΙΜ-β7, BimEL, BimL, Qing are used in the literature as the same, and in the literature This application is intended to be used interchangeably. "The term "oligonucleotide that is specific for ..." or a violent nucleotide that targets ... refers to an oligonucleotide having the following sequence: (1) A stable complex can be formed with a portion of the gene of the dry gene, or (9) a stable duplex can be formed with the portion of the mRNA transcript of the dry gene. The stability of the compound and duplex can be determined by theoretical calculations and/or in vitro analysis. An exemplary analysis for determining the stability of hybridization complexes and duplexes is set forth in the Examples below. The term "target nucleic acid" as used herein encompasses DNA, RNA (including mRNA precursors and mRNA) transcribed from the DNA, and cDNA derived from the RNA, coding sequence, non-coding sequence, sense or antisense polynucleotide. . The specific hybridization of an oligomeric compound to its target nucleic acid interferes with the normal function of the nucleic acid. This modulation of the function of the light nucleic acid with a compound that specifically hybridizes to the nucleic acid is commonly referred to as "antisense." The DNA function to be interfered with includes, for example, copying and transcription. The RNA function to be interfered with includes all important functions, such as shifting of RNA to protein translation sites, translation of proteins from RNA, splicing of RNA to produce one or more mRNA species, and catalytic activity in which RNA can participate or promote. The overall effect of this interference on the function of the target nucleic acid is the regulation of the expression of the encoded product or oligonucleotide. RNA interference ("RNAi") is mediated by double-stranded RNA (dsRNA) molecules with sequence-specific homology to the "target" nucleic acid sequence. In certain embodiments of the invention, the medium has a "small interference" RNA duplex (siRNA) of 5-25 nucleotides. siRNA is derived from the treatment of dsRNA by an RNase enzyme called Dicer. The siRNA duplex product is recruited into a polyprotein siRNA complex (RNA-induced silencing complex) called RISC. Without wishing to be bound by any particular theory, it is believed that RISC can be directed to a target nucleic acid (mRNA is preferred) where the siRNA duplex interacts in a sequence-specific manner to catalyze cleavage in a catalytic manner. Small interfering RNAs useful in the present invention can be synthesized and used according to procedures well known in the art and known to those skilled in the art, 156341. doc • 10-201201819. The small interference (10) VIII used in the method of the invention suitably comprises from about 1 to about 50 nucleotides (nt). In an example of a non-limiting embodiment, the siRNA can comprise from about 5 to about 40 additions, from about 5 to about 30 additions, from about 0 to about 30 nt, from about 15 to about 25 additions, or about 2 Å. _25 nucleotides. Appropriate oligocores (4) (4) are promoted by using computer programs that automatically align nucleic acid sequences and indicate regions of identity or homology. These programs are used to compare the obtained nucleic acid sequences by, for example, searching a database such as GenBank or by sequencing the pCR product. Comparison of nucleic acid sequences from various species allows selection of nucleic acid sequences that show the degree of agreement between appropriate species. In the case of unsequenced genes, Southern blotting is performed to determine the degree of agreement between the target species and other species. Consistency can be roughly measured by performing Southern blots (as is well known in the industry) with varying degrees of stringency. Such procedures allow for the selection of oligonucleotides that exhibit a high degree of complementarity to the dry nucleic acid sequences in the individual to be controlled and a lower degree of complementarity to the corresponding nucleic acid sequences in other species. Those skilled in the art will recognize that a wide range of gene regions suitable for use in the present invention can be selected. "Enzyme RNA" means an RNA molecule with enzymatic activity (Cech, (1988)

American. Med· Assoc. 260, 3030-3035). The enzymatic nucleic acid (ribose _) first acts by binding to the target RNA. The binding is carried out via a target binding portion of an enzymatic nucleic acid that is immediately adjacent to the enzymatic portion of the molecule used to lyse the dry RNA. Thus, the enzymatic nucleic acid first recognizes the crude and then binds to the target RNA via base pairing and acts enzymatically to cleave the target RN A upon binding to the exact site. 156341.doc •11- 201201819 “Bee RNA” means an RNA molecule that mimics the natural binding domain of a ligand. The inducer RNA thus competes with the natural binding stem for binding to a specific ligand. For example, it has been shown that overexpression of HIV transactivation (TAR) RNA can be used as a "slanting" and effective binding to the HIV tat protein, thereby preventing its binding to the TAR sequence encoded in HIV RNA. This is intended to be a specific example. Those skilled in the art will recognize that this is only an example and that other embodiments can be readily derived using techniques generally known in the art. The term "monomer" as used herein generally denotes an oligo which is bonded by a linoleic acid diester bond or an analog thereof to form a size ranging from a plurality of monomer units (for example, about 34) to about several hundred monomer units. Monomeric monomer. Phosphate diacetate linked analogs include phosphorothioate, dithiophosphate, phosphonic acid phosphatide, phosphorothioate, amino phosphate, and the like, as described below. The term "nucleotide" encompasses naturally occurring nucleotides as well as non-naturally occurring nucleotides. Those skilled in the art should be aware that the various nucleotides previously considered to be "unnaturally abundant in J" have since been discovered in nature. Therefore, "nucleic acid" includes not only molecules containing known purine and pyrimidine heterocycles, but also Also included

Its heterocyclic analogs and tautomers. Other types of nucleotide release A cases include molecules of the following parts: adenine, guanine, thymus spray = cytosine, uracil, guanidine, xanthine, diamine.疋7 Side Oxygen-N6-decyl adenine, 7-deazapurine, 7_deaza^ 〇^ ritual horse*呤, N4, N4 ethanol cell" dense bite, N6, N6-ethanol-2 ,6-diaminopurine, T cyanoxyl, 5-(C3-C6)-alkynylcytosine, 5-fluorouracil, / urinary urinary tract, pseudoisomer, 2-hydroxy-5-A 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 The term "nucleotide" is intended to cover each and every such instance as well as analogs and tautomers thereof. Particularly important nucleotides are those containing adenine, black mites, thymine, cytosine, and uracil, which are considered naturally occurring nucleotides associated with therapeutic and diagnostic applications in humans. Nucleotides include native 2'-deoxy and 2,-hydroxy sugars (e.g. ' as described in Kornberg and Baker, DNA Replication, 2nd Edition (Freeman, San Francisco, 1992) and analogs thereof. An "analog" involving a nucleotide comprises a synthetic nucleotide having a modified base moiety and/or a modified sugar moiety (see, for example, as outlined below: Scheit, Nucleotide Analogs, John Wiley, New York, 1980) i

Freier & Altmann, (1997) Nucl. Acid. Res., 25(22), 4429-4443, Toulme, JJ, (2001) Nature Biotechnology 19:17-18 » Manoharan M., (1999) Biochemica et Biophysica Acta 1489:1 17-139 ; Freier SM, (1997) Nucleic Acid Research, 25:4429-4443, Uhlman, E., (2000) Drug Discovery & Development, 3: 203-213, Herdewin P., (2000) Antisense &

Nucleic Acid Drug Dev" 10:297-310); 2'-0, 3'-C linked [3.2.0] bicyclic glucosinolates. These analogs are designed to enhance binding properties (eg 'double stranded' Synthetic nucleotides of bulk or triplex stability, specificity, or the like. As used herein, "hybridization" means the pairing of substantially complementary strands of a polymeric compound. A pairing mechanism involves the recruitment of a hydrogen bond between a complementary nucleoside or a nucleotide base (nucleotide) of a compound chain, which can be Watson-Crick, 156341.doc •13·201201819

Hoiigsteen or reverse Hodgsteen hydrogen bonds. For example, . 'Glandular thyme and thymine are complementary cores formed by hydrogen bond formation, and the anti-sense compound can be "specifically hybridized" when the following conditions are different. The combination of the character and the dry nucleic acid can interfere. The normal function of dry nucleic acids. Or active, and sufficient degree of complementarity exists to avoid the antisense compound nucleic acid sequence under conditions that are expected to undergo specific binding (ie, under physiological conditions in the case of in vivo analysis or therapeutic treatment, and in vitro = analysis) Non-specific binding occurs under conditions in which the analysis is carried out. As used herein, the phrase "stringent hybridization conditions" and "stringent conditions" are the conditions under which the compound of the present invention hybridizes to its dry sequence, but hybridizes with the least number of sequences. The stringent conditions are sequence-dependent and in the same case; In the context of the invention, the "stringent conditions" for hybridization of an oligomeric compound to a dry sequence are determined by the nature and composition of the polymeric compound and its analysis. In general, stringent hybridization conditions include low concentrations (<〇15Μ) salts containing inorganic cations (e.g., ^++ or κ++) (i.e., low ionic strength), temperatures above 2 (rc_25t but below) Oligomeric compound: Tm of the dry sequence complex, and the presence of a denaturant (such as guanamine, dimethyl decylamine, dimethyl sulfoxide) or detergent sodium lauryl sulfate (SDS). For each 1% formamide, the rate of hybridization was reduced by 1.1%. Examples of high stringency hybridization conditions were 0.1X sodium sulphate-sodium citrate buffer (SSC) / 〇.1% (w/v SDS (for 30 minutes at 60 ° C). As used herein, "complementary" refers to the ability to precisely pair between two nucleosides I on one or two oligo-chains. For example, if antisense Compound 156341.doc 201201819 The nucleobase in place is capable of hydrogen bonding with a nucleobase at a position of the target nucleic acid (the target nucleic acid DNA, RNA, or oligonucleotide molecule), then the oligo The position at which hydrogen bonding occurs between the nucleotide and the target nucleic acid is regarded as a complementary position. There is a sufficient number of complementary positions in each molecule to be hydrogenable to each other. When the nucleotides of the knot occupy, the oligomeric compound and other DNA, RNA, or oligonucleotide molecules are complementary to each other. Therefore, "specifically hybridizable" and "complementary" are used to indicate the following terms: a sufficient number of There is a sufficient degree of exact pairing or complementarity in the nucleotide acid to allow for stable and specific binding between the oligomeric compound and the target nucleic acid. It is understood in the art that the oligomeric compound sequence does not require its target nucleic acid sequence 100 to specifically hybridize to it. % complementary. In addition, oligonucleotides may hybridize in one or more segments such that insertion or adjacent segments are not involved in hybridization events (eg, loop structure, mismatch or hairpin structure) The polycompound comprises at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95% of the target region within the target nucleic acid sequence to which it is targeted, or Sequence complementarity of at least about 99. For example, 18 of the 20 nucleotides of the antisense compound are complementary to the target region and the antisense compound that specifically hybridizes will represent 90% complementarity. In this example, the rest Complementary nucleotides may be clustered or dispersed with complementary nucleotides and need not be adjacent to each other or to a complementary nucleotide. Thus, the length is 8 nucleotides and has 4 (four) non-complementary nucleotides (by two The antisense compound flanked by the region in which the target nucleic acid is fully complementary) has a total complementarity of 77.8% with the target nucleic acid and is thus within the scope of the invention. The percent complementarity of the antisense compound having the target nucleic acid region is generally The BLAST program (base local pair 156341.doc 15 201201819 quasi-search tool) and PowerBLAST program are known in the art. The homology, sequence identity or complementarity percentage can be obtained by, for example, Gap program (Wisconsin Sequence Analysis). Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.) was measured using default settings (which use the Smith and Waterman algorithms) (Adv. Appl, Math., (1981) 2, 482-489). The term "thermodynamic melting point (Tm)" as used herein refers to the temperature at which 5 〇 0 / 〇 in an oligonucleotide complementary to a target sequence at a defined ion intensity, pH, and nucleic acid concentration hybridizes to a dry sequence under equilibrium. Generally, for short oligonucleotides (eg, 'having 10 to 50 nucleotides), stringent conditions are those under which the salt concentration is at least about 0.1 to 1.0 MiNa ion at pH 7.0 to 8.3. Concentration (or other salt) and a temperature of at least about 30. Stringent conditions can also be achieved by the addition of destabilizing agents such as formamide. As used herein, "modulation" means the expression of an increase (stimulation) or a decrease (inhibition) of a gene.

When used in the context of a polynucleotide sequence, the term "variant" can encompass a polynucleotide sequence associated with a wild-type gene. This definition may also include, for example, "allele", "splicing", "species" or "polymorphism". The splicing variant may have significant agreement with the reference molecule, but during mRNA processing. The alternate splicing of the exemplants usually has a larger or smaller number of polynucleotides. The corresponding polypeptide may have additional functional domains or no domain. A sequence of nucleotides in which a species-variation system varies between species. Particularly useful in the present invention are variants of the wild type gene product. A variant may be derived from at least one mutation in a nucleic acid sequence and may produce altered mRNA 156341.doc -16 - 201201819 or a polypeptide that may or may not be altered in structure or function. Any given natural or recombinant gene may have no allelic form, with one or many allelic forms. Common mutational changes that result in variants are often attributed to natural deletions, additions, or substitutions of nucleotides. Each of these variations can occur individually or in combination with others in a given sequence one or more times. The resulting polypeptides typically have significant amino acid identity with each other. A polymorphic system changes a polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants may also encompass "single nucleotide polymorphisms" (SNp) or single-sequence mutations in which one base of the polynucleotide sequence is altered. The presence of a SNP can indicate, for example, that a population has a predisposition to disease status (i.e., susceptibility to resistance). The derivatized polynucleotide comprises a nucleic acid that undergoes chemical modification (e.g., hydrogen is replaced by an alkyl group, a thiol group, or an amine group). Derivatives (e.g., derivatized oligonucleotides) can include non-naturally occurring moieties such as altered sugar moieties or intersaccharide linkages. Such exemplary derivatives are phosphorothioates and other sulfur-containing materials known in the art. The derivatized nucleic acid may also contain a label comprising a radioactive nucleotide, an enzyme, a fluorescent agent, a chemiluminescent agent, a chromogen, a substrate, a cofactor, an inhibitor, a magnetic particle, and the like. A derivative polypeptide or peptide is, for example, modified by: glycosylation, pegylation, phosphonium, sulfation, reduction/alkylation, thiolation, chemical coupling, or mild blessing Formalin treatment. The derivative may also be modified to contain a detectable label (directly or indirectly) including, but not limited to, radioisotopes, fluorescent, and enzymatic labels. The term "animal" or "patient" as used herein is intended to include, for example, human 156341.doc -17-201201819, sheep, elk, deer, mule deer, water, mammal, monkey, horse, cow, pig, Goats, dogs, cats, rats, mice, birds, chickens, cockroaches, fish, insects and moths. "Mammals" encompass warm-blooded mammals (such as 'humans and domestic animals) that are usually under medical care. Examples include cats, dogs, horses, cows, and humans, and only humans. "Trating or treatment" encompasses the treatment of a disease state in a mammal and includes (a) in the mammal, in particular in the mammal, which is susceptible to the disease but has not yet been diagnosed with the disease. The state is prevented from occurring in the state; (b) inhibiting the disease state, for example, preventing its development; and/or (c) slowing the disease state, for example, reducing the disease state until the desired endpoint is reached. Treatment also includes amelioration of the symptoms of the disease (e. g., pain relief or discomfort), wherein the improvement may or may not directly affect the disease (e.g., cause, infection, performance, etc.). As used herein, "cancer" refers to all types of cancer or neoplasms or malignancies found in mammals including, but not limited to, leukemia, lymphoma, melanoma, carcinoma, and sarcoma. Cancer itself manifests itself as a "tumor" or tissue that includes malignant cells of cancer. Examples of tumors include sarcomas and carcinomas such as, but not limited to, fibrosarcoma, mucinous sarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphatic endothelial sarcoma, synovium Tumor, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland cancer, Sebaceous gland cancer, papillary carcinoma, papillary adenocarcinoma, 156341.doc 201201819 cystadenocarcinoma, medullary carcinoma, bronchial ductal carcinoma, renal cell carcinoma, hepatocellular carcinoma, biliary cancer, choriocarcinoma, seminoma, Embryonic cancer, Wilms' tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma Tumor, ependymoma, pineal tumor, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, and retinoblastoma. Additional cancers treatable by the compositions disclosed herein include, but are not limited to, for example, Hodgkin's disease, non-Hodgkin's lymphoma (N〇n_H〇dgkin, s Lymph〇) Ma), multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, rhabdomyosarcoma, essential thrombocytosis, primary macroglobulinemia, small cell lung tumor, primary brain tumor, Gastric cancer, colon cancer, malignant pancreatic insulinoma, recorded carcinoid, bladder cancer, gastric cancer, pre-malignant skin damage, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary cancer, malignant hypercalcemia Disease, cervical cancer, endometrial cancer, adrenal cortical cancer, and prostate cancer. The term "apoptosis" means programmed cell death. It also means "cell death" or "targeted cell death" which occurs by administering an antisense oligonucleotide as described herein for treatment or reduction of uncontrolled cell growth and/or normal apoptosis and/or In the case of a disease or condition associated with a malfunction of the apoptotic pathway. As used herein, "neurological disease or condition" refers to the nervous system and/or the visual system, any of the first diseases or conditions. "Nervous diseases or conditions" include the middle sacral nervous system (brain, brainstem and cerebellum), the peripheral nervous system (including Gushen, II), and the autonomic nervous system (156341.doc in both the central and peripheral nervous systems - 19-201201819 Part) A disease or condition. Neurological diseases or conditions include, but are not limited to, acquired epilepsy aphasia; acute disseminated encephalomyelitis; adrenal leukodystrophy; age-related macular degeneration; corpus callosum dysplasia; aphasia; Aicardi syndrome; Alexander disease 'A pers, disease'; Alzheimer's disease; vascular dementia; amyotrophic lateral sclerosis, · no brain malformation; Angelman syndrome; chlorocholine disease; dystrophy; aphasia; apraxia; arachnoid cyst, arachnoiditis, Anr〇nl Chiari malformation; arteriovenous malformation; Asperger syndrome; ataxia telangiectasia; attention deficit with hyperactivity disorder; autism; autonomic dysfunction; back pain; Batten disease; Bechette Behcet's disease; Bell's palsy; benign essential hernia; benign local muscle atrophy; benign intracranial pressure, negative Swann disease (Binswanger, s Disease) Fallen' Bloc-Su 4 Beck syndrome (Bi〇ch Sulzberger syndrome); Brachial plexus injury; Brain abscess; Brain injury; Brain tumor (including Glioblastoma multiforme); Spinal cord tumor; Brown-Seccar syndrome (Br〇wn_

Sequard syndrome), Canavan disease; carpal tunnel syndrome; carcinogenic neuralgia; central pain syndrome; central medullary cerebral ischemic; head condition; cerebral aneurysm; cerebral arteriosclerosis; brain atrophy; Sexual giant disease; cerebral palsy; Charc〇t_ Made-Tooth disease; chemotherapy-induced neuropathy and neuropathic pain, Chiari malformation; chorea; chronic inflammation Sexual 156341.doc -20- 201201819 Demyelinating polyneuropathy; chronic pain; chronic regional pain syndrome 'Coffin Lowry syndrome; coma, including persistent vegetative state; congenital facial paralysis; cortex Basal degeneration; cranial arteritis 'cranial suture early closure, Creutzfeidt-Jakob disease; accumulative traumatic disorder, Cushing's Syndrome; giant cell inclusion disease; cytomegalovirus infection; - Dance foot syndrome, Dandy Walker syndrome; Dawson disease; De Morsier's syndrome; Dejerine_Klumke palsy, dementia; dermatomyositis; diabetic neuropathy; diffuse sclerosis; autonomic dysfunction; difficulty in writing; dyslexia; dystonia; early childhood epilepsy encephalopathy; Syndrome; encephalitis; cerebral palsy; brain three neuromuscular disease; epilepsy; Obo paralysis (7) 吒, palsy); idiopathic tremor; Fabry's disease; Fahr syndrome ); fainting; familial spastic paralysis; febrile seizures; Fisher syndrome; Friedreich's ataxia; frontotemporal dementia and other "tau lesions"; Gaucher's disease; Gerstmann's syndrome; giant cell arteritis; giant cell inclusion disease; globular cell leukodystrophy; Ge-Ba_Mu &> w 〇-jiu Guillain-Barre syndrome; HTLV-1 associated myelopathy; Hallervorden-Spatz disease; head injury. head, wide. semi-ma facial paralysis; hereditary spastic paraplegia; Pathogenic polyneuritis-like sputum dislocation; ear herpes zoster; herpes zoster; Pingshan syndrome (Hiraya 156341.doc -21 · 201201819 syndrome); HIV-related dementia and neuropathy (and neurological manifestations of AIDS) ), forebrain non-cracking malformation; Huntington's disease and other polyglutamine indole repeat disease; water-free no brain malformation; hydrocephalus; hypercortisolism; hypoxia; immune-mediated encephalomyelitis; Myositis; pigment disorders; infant phytanic acid storage disease; infantile refsum disease; infantile spasm; inflammatory myopathy; intracranial cyst; intracranial hypertension; jubbert syndrome (Joubert Syndrome); Keams-Sayre syndrome; Kennedy disease, Kinsboume syndrome; Klippel Feil syndrome; Klaper Krabbe disease, Kugelberg-Welander disease, Kuru disease; Lafora disease; Lang-Eaton myast syndrome (Lambert-Eaton myast) Henic syndrome); Landau-Kleffner syndrome; lateral medullary (Wallenberg) syndrome; learning disability; Leigh's disease; Lennox-Gasto Lennox-Gustaut syndrome; Lesch-Nyhan syndrome, leukodystrophy; Lewy body dementia; no cerebral gyrus; atresia syndrome; Lu-grid disease (L〇u Gehrig's disease) (ie, motor neuron disease or amyotrophic lateral sclerosis) ' lumbar disc disease; Lyme disease - neurological sequelae; Machado-Joseph disease; brain Hypertrophy; giant brain; Melkersson-Rosenthal syndrome; Menieres disease; meningitis; Menkes disease; 156341.doc •22- 201201819 Miller. Faxi granule muscle metasomal leukodystrophy; microcephaly; Miller Fisher syndrome; small stroke, Mobius syndrome; single limb muscle atrophy; motor neuron Abnormal vascular network Mucopolysaccharidosis; multiple infarct dementia; multifocal motor neuropathy; multiple sclerosis and other demyelinating disorders; multiple systemic atrophy with location hypotension; muscular dystrophy; myasthenia gravis; Sexual sclerosis; myoclonic encephalopathy; myoclonus, myopathy; myotonia; narcolepsy; neurofibromatosis, disease; neuroleptic malignant syndrome; neurological manifestations of AIDS; neurological sequelae of lupus; neuromuscular rigidity Neuronal wax-like lipofuscin; abnormal brain neuron migration, Niemann-PickdiSease; O'Sullivan's disease (0'SUllivan_McLe〇d syndrome); occipital neuralgia; recessive Spinal canal insufficiency sequence; Daejeon syndrome (10) dynasty syndrome; platycotina cerebral atrophy; strabismic cerebral palsy; optic neuritis; orthostatic hypotension; overuse syndrome; paresthesia; neurodegenerative diseases or disorders (Parkinson, s disease, Huntington's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALs), dementia, multiple sclerosis and Other diseases and conditions related to neuronal cell death); congenital accessory myotonia; paraneoplastic disease; paroxysmal attack; Parry Romberg syndr〇me; Pey-Mei's disease Pelizaeus-Merzbacher disease); peripheral paralysis; peripheral neuropathy; painful neuropathy and neuropathic pain; persistent vegetative state; systemic developmental delay; optical sneezing reflex; (4) acid storage disease; Pick's disease; Kneading; pituitary tumor; polymyositis, · brain penetrating aberration I56341.doc •23· 201201819 shape; polio late syndrome; post-herpetic neuralgia; salty encephalomyelitis; orthostatic hypotension; Wei Er's syndrome (Prader_Wi out syndrome); primary lateral sclerosis; prion disease; progressive one-sided atrophy; progressive multifocal leukoencephalopathy; progressive sclerotic gray matter atrophy; progressive supranuclear palsy; Brain tumor; Ramsay-Hunt Syndr〇me (types I and II); Rasmussen's encephalitis; reflex sympathetic dystrophy Refsum disease; repetitive motor disorder; repetitive compression injury; restless leg syndrome; retrovirus-associated myelopathy; Rett syndrome; Reye syndrome s syndrome); Saint Vitus dance; Sandhoff disease; Schilder's disease; cerebral palsy; clear septum-optical dysplasia; scare infant syndrome; Shear therapy; Shy-Drager syndrome; Sjogren's syndrome; sleep apnea; Soto's syndrome; spasticity; spina bifida; spinal cord injury; Spinal muscular atrophy; Stiff Pers〇n syndrome; Stroke, Sturge Weber syndrome' subacute sclerosing encephalitis; subcortical arteriosclerotic encephalopathy, Sydenham chorea; syncope; syringomyelia; tardive dyskinesia; Tay_Sachs disease; temporal arteritis; spinal cord syndrome; Tom Th〇msen disease; thoracic outlet syndrome; Tic Douloureux; T〇dd, s paralysis; hyperactive slang 156341.doc • 24 - 201201819 candidate; transient Cerebral ischemic attack; disseminated spongiform encephalopathy; transverse myelitis; traumatic brain injury; trembling; trigeminal neuralgia; tropical spastic paraplegia; tuberous sclerosis; vascular dementia (multiple infarct dementia); I Yan' contains temporal arteritis, V〇n Hippel-Lindau disease; Wallenberg's syndrome; Werdnig-Hoffman disease; Wei West syndrome; cervical spine bruising; Williams syndrome, wildon's disease; and Zellweger syndrome. A "proliferative disease or condition" includes, but is not limited to, a hematopoietic neoplastic disorder involving a cell derived from a proliferative/neoplastic cell, or a precursor cell thereof, which is also derived from a bone marrow-like, lymphoid or erythrocyte lineage. Such diseases include, but are not limited to, erythroblastic leukemia, acute anterior osteocholine leukemia (APML), chronic myeloid leukemia (Cml), lymphoid malignancies (including but not limited to acute lymphoblastic leukemia (ALL), which include B - lineage ALL and T-lineage ALL), chronic lymphocytic leukemia (CLL), lymphoblastic leukemia (PLL), hairy cell leukemia (HLL), and Waldenstrom's macroglobulinemia (WM) ). Other forms of malignant lymphoma include, but are not limited to, non-Hodgkin's lymphoma and its variants, peripheral T-cell lymphoma, adult tau cell leukemia/lymphoma (ATL), cutaneous T-cell lymphoma (CTCL), giant granules Lymphocytic leukemia (LGF), Hodgkin's disease, and Reed_Sternberg disease ° "Blood disease or condition" includes diseases affecting hematopoietic cells or tissues, 156341.doc -25- 201201819 Or condition. A blood disorder contains a disease, disorder, or condition associated with an abnormal blood content or function. Examples of blood disorders include conditions derived from treatment of bone marrow or cancer chemotherapy, such as -, hemorrhagic sputum, hemolytic anemia, aplastic anemia, sputum anemia, iron granulocyte anemia, and chronic infection ( For example, cancer, trypanosomiasis, HIV, hepatitis virus or other viruses) anemia, bone marrow anemia caused by bone marrow deficiency, kidney failure caused by anemia, anemia, polycythemia, infectious mononucleosis (IM), acute non-lymphocytic leukemia (ANLL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), acute granulocyte-monocytic leukemia (AMM〇L), polycythemia vera, Lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia, Wilms, stum〇r, Ewing's sarc〇ma, retinoblastoma, hemophilia, and Blood tests for increased risk associated with illness, running therapy, thalassemia, antibody-induced conditions (such as transfusion reactions and erythroblastosis), red blood cell machinery Trauma (eg, microangiopathic hemolytic anemia, thrombotic thrombocytopenic purpura, and disseminated intravascular coagulation), infection by parasites (eg, cancer), chemical damage from, for example, lead poisoning, and spleen Hyperfunction. Lymphatic diseases include, but are not limited to, lymphadenitis, lymphatic expansion, lymphangitis, lymphedema, lymphocystosis, lymphoproliferative disorders, cutaneous mucosal lymph node syndrome, reticuloendothelial proliferation, spleen disease, thymic hyperplasia, thymic neoplasms, Tuberculosis, lymph nodes, pseudolymphoma, and lymphoid abnormalities. The conditions of the lymphoid jk system include, but are not limited to, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, and reactive lymphoid group 15634l.doc 26·201201819 woven hyperplasia. Polynuclear SllL and Nucleic Acid Compositions and Molecules and: In one embodiment, the target comprises a nucleic acid sequence of BCL2 11 (BCL2L11) comprising (without limitation) a sense and/or antisense associated with BCL2L11 Coding and / or coding sequences.

Members of the Bcl-2 family of proteins are structurally similar but have different activities. This family of proteins is divided into members of the "pro-apoptotic" family (ie, Bax, Bak, Bok) and members of the "promoting survival" family (ie, Bci_2). , Bcl-XL, Bcl-w) ^ The former can be further subdivided into their proteins containing the bh3 domain and their lack of BH3 domain. It is believed that the BH3 domain facilitates the direct binding of such proteins to "promoting" proteins, thereby inhibiting "promoting" activity and promoting cell death or apoptosis. The human BCL2L11 locus is located on chromosome 2qi 3, which encodes a protein of 198 amino acids that is structurally and functionally related to the only BH3 class of pro-apoptotic BCL2 family members. Genes are frequently mutated in different human tumors, resulting in loss of BCL2L11 activity. BCL2L11 expression is induced by a range of apoptosis stimuli (e.g., deletion of growth factors/cytokines, ionizing radiation, and cytotoxic peptides). In one embodiment, an antisense oligonucleotide is used to prevent or treat a disease or condition associated with a member of the BCL2L11 family. Exemplary BCL2 11 (BCL2L11) mediated diseases and conditions that can be used in cell/tissue treatment derived from stem cell regeneration using antisense compounds include: cancer, abnormal cells > weekly death, proliferative diseases or disorders, and grain lines A disease or condition associated with a path of apoptosis in a body, a leukemia, an autoimmune disease or condition, a disease or condition associated with an epidemic damage, infection, hyperproliferative disorder, reproductive tissue Hyperproliferation (eg uterine cancer, testicular cancer and (4) cancer, endometriosis, squamous cervix, glandular epithelial cancer, etc.), neurological diseases or conditions, developmental disorders associated with developing embryos, blood Disease or condition, tissue homeostasis, rheumatoid arthritis, septicemia, retinal ganglion cell death 'liver disease and kidney disease. In one embodiment, BCL2L11 is modulated by one or more antisense oligonucleotides in a patient in need thereof to prevent or treat any of the abnormalities, functions, and activities of BCL2LU (as compared to a normal control group) A disease or condition. In one embodiment, the oligonucleotide is specific for a polynucleotide of BCL2L11, which comprises, without limitation, a non-coding region; a BCL2L11 target comprising a variant of BCL2L11; a mutant of BCL2L11 comprising a SNP; a non-BCL2L11 Coding sequences; alleles, fragments, and the like. Preferably, the oligonucleotide is an antisense RNA molecule. According to an embodiment of the invention, the target nucleic acid molecule is not limited to BCL2L11 polynucleic acid but extends to any of the isoforms, receptors, homologs, non-coding regions and the like of BCL2L11. In one embodiment, the oligonucleotide targets a natural antisense sequence (a natural sequence encoding a non-coding region) of a BCL2L11 target, including, without limitation, variants, alleles, homologs, mutants thereof, Derivatives, fragments and complementary sequences. Preferably, the oligonucleotide is an antisense RNA or DNA molecule. In one embodiment, the oligomeric compounds of the invention also comprise variants in which different bases are present at one or more nucleotide positions in the compound. For example, 156341.doc •28· 201201819 言 “If the first nucleotide is adenine, a variant containing a chestnut, guanosine, cytidine or other natural or non-natural nucleotide at this position may be produced. This can occur anywhere in the antisense compound. The compounds are then tested using the methods described herein to determine their ability to inhibit the performance of the target nucleic acid. In some embodiments, the homology, sequence identity or complementarity between the antisense compound and the target is from about 50% to about 60%. In some embodiments, homology, sequence identity, or complementarity is from about 6% to about 7%. In some embodiments, 'homology, sequence identity, or complementarity is about 70〇/〇 to About 80%. In some embodiments, the homology 'sequence identity or complementarity is from about 80% to about 90%. In some embodiments, the homology, sequence identity, or complementarity is about 90%, about 92%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 1〇〇%. An antisense compound can specifically hybridize in the following situations: binding of the compound to the dry nucleic acid can interfere with the normal function of the target nucleic acid resulting in loss of activity, and a sufficient degree of complementarity exists to avoid antisense compounds and non-dry nucleic acid sequences from occurring as desired. Non-specific binding occurs under conditions of specific binding. Such conditions include (i.e., physiological conditions in the case of in vivo analysis or therapeutic treatment, and conditions under which the analysis is performed in the context of in vitro analysis. An antisense compound (whether DNA, RNA, chimeric compound, or substituted compound, etc.) can specifically hybridize in the following situations: binding of the compound to a target DNA or RNA molecule can interfere with the normal function of the target DNA or RNA. Loss of utility, and sufficient degree of complementarity to avoid antisense compounds and non-target sequences under conditions that are expected to specifically bind (ie, under physiological conditions in the case of in vivo analysis or therapeutic treatment, or in vivo) 156341.doc • 29-201201819 Under the conditions of performing an analysis in the case of an external analysis, the occurrence of non-specific binding β In one embodiment, 'undirected BCL2L11 (including (not limited to) identification and expansion using, for example, PCR, hybridization, etc. Antisense sequences, such as SEq ID Ν〇: one or more of the sequences shown in 2 and 3, and the like, can modulate the performance or function of bcl2L 11. In one embodiment, the performance or function is up-regulated compared to the control group. In one embodiment, the performance or function is downregulated as compared to a control group. In one embodiment, the oligonucleotide comprises a nucleic acid sequence as set forth in SEQ ID NOS: 4 to 13, comprising an antisense sequence which is identified and expanded using, for example, PCR, hybridization or the like. Such oligonucleotides may include one or more modified nucleotides' shorter or longer fragments, modified linkages, and the like. Examples of the modified bond or the internucleoside linkage include a phosphorothioate, a dithiophosphate or the like. In the case of consistent fineness, 'nuclear acid includes a derivative. Phosphorus derivatives (or modified phosphate groups) that can be attached to a sugar or a sugar analog moiety of a modified oligonucleotide of the invention can be monophosphates, diphosphates, tri-salts, alkyl phosphates Base esters, alkyl phosphates, phosphorothioates, and the like. The preparation of the above citrate analogs, as well as the methods of their entry into nucleotides, modified nucleotides and nucleotides are also known per se and need not be set forth herein. Those skilled in the art also utilize the specificity and sensitivity of antisense oligonucleotides in therapeutic applications. Antisense oligonucleotides are used as therapeutic moieties to treat disease states in animals and humans. Antisense nucleotides have been safely and efficiently administered to humans and many clinical trials are currently being performed. It has thus been determined that oligonucleosides can be a useful therapeutic modality that can be used in the treatment of cells, tissues and animals, especially humans. In an embodiment of the invention, an oligo antisense compound, particularly an oligonucleotide, 156341.doc 201201819 binds to a dry nucleic acid molecule and modulates the expression and/or function of the molecule encoded by the target gene. DNA functions to be interfered with include, for example, replication and transcription. The RNA function to be interfered with includes all important functions, such as shifting of RNA to protein translation sites, translation of proteins from RNA, splicing of RNA to produce one or more mRNA species, and catalytic activity in which RNA can participate or promote . The function can be adjusted or suppressed depending on the desired function. An antisense compound comprises an antisense oligomeric compound, an antisense oligonucleotide, an external leader sequence (EGS) nucleus nucleotide, an alternative splicing, a primer, a probe, and other oligomeric compounds that hybridize to at least a portion of the target nucleic acid. Thus, the compounds can be introduced in the form of single-stranded, double-stranded, partially single-stranded, or cyclic oligomeric compounds. In the context of the present invention, the drying of an antisense compound to a particular nucleic acid molecule can be a multi-step process. This process often begins with the identification of target nucleic acids that regulate function. For example, the target nucleic acid can be a nucleic acid molecule that exhibits a cellular gene (or mRNA transcribed from the gene) associated with a particular disorder or disease state or from an infectious agent. In the present invention, the target nucleic acid encodes bcl2 u (BCL2L11). The targeting process also often includes determining at least one target region, region, or site within which the antisense interaction occurs within the target nucleic acid to produce a desired effect (e.g., modulate expression). Within the context of the present invention, the term "region" is defined as a portion of a target nucleic acid having at least one identifiable structure, function, or property. There is a segment within the region of the target nucleic acid. A "segment" is defined as a small or sub-portion of a region within a target nucleic acid. The "site" used in the present invention is defined as the position within the dry nucleic acid. 156341.d〇) 31 201201819 In one embodiment, the antisense oligonucleotide binds to a natural antisense sequence of BCL2 11 (BCL2L11) and regulates the expression and/or function of BCL2L11 (SEQ ID NO: 1). An example of an antisense sequence comprising SEqNO: 2 to 13 〇 In one embodiment, the 'antisense oligonucleotide binds to one or more of the BCL2 11 (BCL2L11)-like polynucleotides and modulates the expression of Bcl2L1 1 and / or function. The segment includes at least 5 contiguous nucleotides of a BCL2L11 sense or antisense polynucleotide. In one embodiment, the antisense oligonucleotide is specific for the natural antisense sequence of BCL2L11, and binding of the oligonucleotide to the natural antisense sequence of BCL2L11 modulates the expression and/or function of BCL2L11. In one embodiment, the oligonucleotide compound comprises the sequence as set forth in SEQ ID NOS: 4 to 13, and the antisense sequence identified and expanded using, for example, PCR, hybridization, or the like. Such oligonucleotides may include one or more modified nucleotides, shorter or longer fragments, modified linkages, and the like. Examples of modified bonds or internucleotide linkages include phosphorothioates, dithiophosphates or the like. In one embodiment, the nucleotide comprises a phosphorus derivative. A destructive derivative (or modified acid ester group) that can be attached to a sugar or a sugar analog moiety of a modified oligonucleotide of the invention can be a monostearate, a di-salt, a triphosphate, Alkyl phosphates, alkyl phosphates, phosphorothioates, and the like. The preparation of the above-described acid ester analogs, and their incorporation into nucleotides, modified nucleotides and oligonucleotides are also known per se and need not be set forth herein. As is known in the art, since the translation initiation codon is usually 5'-AUG (in the transcription of mRNA molecules; in the corresponding DNA molecule is 5'-ATG), the translation is activated 156341.doc -32· 201201819 It is "AUG Codon" - "Start Codon" or "Bagua ^ Start Codon". A few genes have a translation initiation codon with an rna sequence of 5, -GUG, 51-UUG or 51-CUG, and it has been shown that 5, _AUA, ACG and 5'-CUG can function in vivo. Therefore, even if the starting amino acid is usually methionine (in eukaryotes) or methotrexate (in prokaryotes) in each case, the terms "translation start codon" and "start" Codons can also encompass a wide variety of codon sequences. Eukaryotes and prokaryotic genes can have two or more alternative initiation codons, either of which can be preferentially used in a particular cell type or tissue. Or a translation start under a specific condition group. In the context of the present invention, "start codon" and "translation start codon" refer to one or more codons used to initiate translation of mRNA from a gene encoding a gene encoding BCL2 11 (BCL2L11) in vivo. , regardless of the sequence of such codons. The gene translation stop codon (or "stop codon") can have three sequences (ie, 5·· UAA, 5'-UAG, and 5I-UGA, and the corresponding DNA sequences are 5, _TAA, 5'-TAG, and One of 5'-TGA). The "delta start codon region" and "translation start codon region" mean that the mRNA or gene covers from about 25 to about 50 in either direction from the translation initiation codon (ie, 5 or 3'). Part of a nearby nucleotide. Similarly, the stop of the code sub-region and the "transition stop codon region" means that the mRNA or gene covers from about 25 to about 50 in either direction from the translation stop codon (ie, 5' or 3'). Part of a nearby nucleotide. Therefore, the start codon region (or "translation start codon region") and "catch-in codon region" (or "transfer stop codon region") are 156341.doc -33-201201819 invented antisense compounds Effectively work in all areas. An open reading frame (ORF) or "coding region" known in the art refers to the region between the translation initiation codon and the translation stop codon, which is also an area that can be effectively targeted. Within the context of the present invention, the targeting region encompasses the intragenic region of the translational start or stop codon of the open reading frame (ORF) of the gene. Another target region comprises the 5|non-translated region (5, UTR) known in the art, which refers to the portion of the mRNA in the direction from the translation initiation codon 5, and thus comprises the 5' capping site of the mRNA. And translating the nucleotides (or corresponding nucleotides in the gene) between the initiation codons. Another target region comprises the 3, non-translated region (rUTR) known in the art, which refers to the portion of the mRNA in the direction from the translation stop codon 3, and thus contains the translation stop codon and the 3' end of the mRNA. Between the nucleotides (or the corresponding nucleotide nucleoside mRNAi5, the addition of the Nb-based thiolation of the 5'-most residue of the mRNA via the 5'-5' triphosphate linkage The guanosine residue. mRNA25, the capped region is considered to contain the y capping structure itself and 5 nucleotides before the capping site. Another dry region of the invention is the 5' capped region. Nuclear biological mRNA transcripts can be translated directly, but many eukaryotic mRNA transcripts contain one or more regions that are cut from the transcript prior to translation and are referred to as "introns." The remaining (and thus translated) regions are called "Exons" and spliced together to form a continuous mRNA sequence. In one embodiment, a targeted splice site (i.e., an intron-exon node or an exon-intron node) is particularly Can be used for abnormal splicing related to disease' or excessive production of specific splicing products related to disease In the case of rearrangement or loss of production 156341.doc

-34- 201201819 Another embodiment of the abnormal fusion junction node target site. An mRNA transcript produced by splicing two (or more) mRNAs from different gene sources is referred to as a "fusion transcript." Antisense compounds that target, for example, DNA or mRNA precursors can be used to effectively render the undirected intron. In one embodiment, an antisense oligonucleotide binds to a coding and/or non-coding region of a target polynucleotide and modulates the expression and/or function of the target molecule. In one embodiment, the antisense oligonucleotide binds to a natural antisense polynucleotide and modulates the expression and/or function of the target molecule. In one embodiment, the antisense oligonucleotide binds to a sense polynucleotide and modulates the expression and/or function of the target molecule. Alternative RNA transcripts can be produced from the same genomic region of DNA. Such alternative transcripts are commonly referred to as "variants." More specifically, "mRNA precursor variants" are transcripts produced from the same genomic DNA that differ from the other transcripts produced from the same genomic DNA in terms of starting or stopping positions and contain introns and Proton sequence. When one or more exons or intron regions, or portions thereof, are cleaved during splicing, the mRNA precursor variants produce smaller "mRNA variants". Thus, mRNA variants of processed mRNA precursor variants and each unique mRNA precursor variant must always produce a unique mRNA variant by splicing. These mRNA variants are also referred to as "alternate splice variants". If the mRNA precursor variant is not spliced, the mRNA precursor variant is identical to the mRNA variant. Variants can be generated via the use of alternating signals that initiate or stop transcription. The mRNA precursor and mRNA may have more than one initiation codon or stop 156341.doc -35 - 201201819 MM. Variants derived from the secret precursor or mRNA using an alternative initiation codon are referred to as mRNA precursors or "alternative initiation variants" of the capture. He used an alternative stop Maggie M丄 to treat the transcript in the code called the “alternative stop variant” of the mRNA precursor or mRNA. A specific type of alternative stop variant is a "P〇lyA variant" in which multiple transcripts are derived from an alternative to the transcription machine for one of the "p〇lyA stop signals", resulting in A transcript that terminates at a unique polyA site. Within the context of the present invention, the types of variants described herein are also examples of target nucleic acids. The position on the target nucleic acid that hybridizes to the antisense compound is defined at least as a portion of the target region that is 5 nucleotides long that is targeted by the active antisense compound. Although specific sequences of certain exemplary target segments are set forth herein, those skilled in the art will recognize that such sequences are used to exemplify and describe particular embodiments within the scope of the invention. Those skilled in the art can readily identify other starting segments in accordance with the present disclosure. It is believed that a dry segment of 5-1 00 nucleotides in length and comprising at least five (5) contiguous nucleotides selected from the preferred interpretive light segments is also suitable for extension to 0. The light segment may comprise DNA or An RNA sequence comprising at least 5 contiguous nucleotides from the 5'-end of one of the preferred interpretable target segments (the remaining nucleotides are consecutive segments of the same DNA or RNA, which are immediately adjacent to the target The upstream of the 5'-end of the segment begins and continues until the DNA or RNA contains from about 5 to about 1 nucleotide. Likewise, a preferred target segment is represented as a DNA or RNA sequence which comprises at least 5 contiguous nucleotides from the 3, _ terminus of one of the preferred interpretable target segments (the remaining nucleotides are the same DNA or In the continuation of the I56341.doc •36·201201819, the RNA starts immediately downstream of the 3,_end of the target segment and continues until the DNA or RNA contains about 5 to about 1 nucleotide. Those who are familiar with this technique can identify other better target segments without undue experimentation. An antisense compound that is sufficiently complementary (i.e., sufficiently hybridized and sufficiently specific) can be selected to achieve the desired effect, either after the vacant region, segment or site. In an embodiment of the invention, the oligonucleotide binds to an antisense strand of a particular target. The oligonucleotides are at least 5 nucleotides in length and can be synthesized such that each oligonucleotide is dried to the overlapping sequence such that the oligo (4) is synthesized to cover the entire length of the dry polyphosphate. The code also includes both coded and non-coded areas. In one embodiment, the antisense oligonucleotide preferably targets a specific nucleic acid. The dry direction of the anti-sense δ to the 疋 nucleic acid is a multi-step process. This process often begins with the identification of the nucleic acid sequence that regulates the function. This may, for example, be a cellular gene (or mRNA transcribed from a gene) associated with a particular disorder or condition, or a non-coding polynucleotide (e.g., non-coding RNA (ncRNA)). RNA can be classified into (丨) messenger 1^8 (mRNA), which translates into protein; and (2) non-protein-encoding RNA (ncRNA). ncRNAs include microRNAs, antisense transcripts, and other transcriptional units that contain high-density stop codons and lack any extensive "open reading frame". Many ncRNAs appear to start at the 3, non-translated region (3 UTR) of the locus of the protein-encoding locus. ncRNAs are generally rare and at least half of the ncRNAs that have been sequenced by the fant〇m association appear to be unpolyadenylated. For the obvious reasons, most of them focus on the polyadenylation mRNA that has been processed and exported to the cytoplasm 156341.doc •37·201201819. Recently, the group showing non-polyadenylated nuclear rna may be extremely large, and many of these transcripts are derived from the so-called intergenic region. The mechanism by which ncRNA regulates gene expression is linked to the base of the target transcript. RNAs that function by base pairing can be grouped into: (1) cis-encoded RNA, which encodes at the same genetic position at which it functions, but encodes at the opposite strand of rna and thus shows complete complementarity with its target And (2) trans-encoded RNA, which encodes at a different chromosomal location than the RNA it functions with and typically does not exhibit full base pairing potential with its target. Without wishing to be bound by theory, the interference of antisense oligonucleotides described herein with antisense polynucleotides can alter the performance of the corresponding sense messenger RNA. However, this modulation may be uncoordinated (anti-sense knockdown causes messenger rna to increase) or co-regulation (antisense weakening results in concomitant messenger RNA reduction). In such cases, the antisense oligonucleotide can target overlapping or non-overlapping portions of the antisense transcript to create a weakening or sequestration. Both coding and non-coding antisense transcripts can be targeted in the same manner, and any species can modulate the corresponding sense transcript in a coordinated or uncoordinated manner. The strategy employed in identifying new oligonucleotides for use with a target can be based on any other means of weakening the antisense RNA transcript by antisense oligonucleotides or modulating the desired lightness. Gray enough 7. In the case of uncoordinated regulation, weakening antisense transcripts will increase the performance of the conventional (sense) gene. If the latter gene encodes a known or putative drug target' then it is conceivable to attenuate its antisense counterpart to mimic the effects of a receptor agonist or an enzyme stimulator. Strategy 2: In the case of coordinated regulation, antisense and sense transcripts can be simultaneously weakened and thus synergistically reduced (negative) gene expression. For example, 156341.doc •38· 201201819 Tongue 'If antisense oligonucleotides are used to achieve weakening, this strategy can be used to apply antisense oligonuclear acid to the sense transcript and the corresponding antisense Another antisense oligonucleotide of the transcript, or a single energy symmetric antisense oligonucleotide that simultaneously targets overlapping sense and antisense transcripts. According to the invention, the antisense compound comprises an antisense oligonucleotide, a ribozyme, and the like. A leader sequence (EGS) oligonucleotide, an siRNA compound, a mono- or double-stranded RNA interference (RNAi) compound (e.g., an siRNA compound), and other oligomeric compounds that hybridize to at least a portion of the target nucleic acid and modulate its function. Thus, it can be DNA, RNA, DNA-like, RNA-like, or a mixture thereof, or can be a mimetic of one or more of such substances. The compounds may be single-stranded, double-stranded, cyclic or hairpin-type polycondensation compounds and may contain structural elements such as internal or terminal expansion, mismatch or loop. Antisense compounds are typically made in a linear form' but may be joined or otherwise formed into a cyclic and/or branched form. Antisense compounds can comprise, for example, constructs that are hybridized to form two strands of a fully or partially double-stranded compound, or that have sufficient self-complementary steepness to hybridize to form a fully or partially double-stranded compound. The two keys can be internally connected to produce a free 3, or 5, end, or can be joined to form a continuous hairpin structure or loop. The hairpin type structure may have a depending portion at the end of the 5 or Γ to extend the long chain characteristics of the single ▲ person 4. The double-stranded compound may have an overhang at the end as needed. Other modifications may include attachment to one end, a selected nuclear acid position, a sugar position, or a coupling group attached to an internucleoside linkage. Another option is two chains It can be linked by a non-nuclear moiety or a linker group. If it is formed by only one stop hammer and eight m bonds, the dsRNA can be self-complementary hairpin type molecular form, JL self-feeding 4匕, 斤 to form a duplex. Therefore, dsRNA can be 156341.doc.39· 201201819 c^NA mai t. It can specifically regulate (4) expression by stably expressing N 々 in the transgenic cell line, ^, In a differential embodiment, the gene expression or function is up-regulated. If two bonds are formed, or a single-stranded form of the auto-complementary hairpin-type molecule (self-folding to form a double bond), the two chains (or A region in which a duplex is formed in a single strand) is a complementary RNA strand that is base-paired in a partial _Cnck manner. After introduction into a system, the compound of the present invention can initiate the action of _ or a plurality of enzymes or structural proteins to effect cleavage of the target nucleic acid. Or other modifications may be made via a mechanism based on occupancy. And t, the nucleic acid (including the nucleus acid) can be described as "DNA-like" (that is, usually with one or more 2,_deoxy sugar and (usually) T instead of U base) or "RNA-like" (ie, usually having one or more 2,-hydroxy or 2,-modified sugars and (usually}u instead of tau bases). Nucleic acid helices may employ more than one type of structure, most commonly Α type and type 8 It is believed that 'generally, the nucleotides with a Β-like structure are "DNA-like" and those with a Α-like structure are "like rNA" ^ in some (intersected) embodiments The compounds may contain A- and B-type regions. In one embodiment, the desired oligonucleotide or antisense compound comprises at least one of the following: antisense RNA, antisense DNA, chimeric antisense oligonucleotides , including modified antisense oligonucleotides, interfering RNA (RNAi), short stem

Scrambled RNA (siRNA); small interfering RNA (miRNA); small temporal RNA (stRNA); or short hairpin RNA (shRNA); small RNA-induced gene activation (RNAa); small activating RNA (saRNA), or a combination thereof . dsRNA also activates gene expression, a mechanism known as "small RNA-induced gene activation" or RNAa. The dsRNA-inducible phase targeting the gene promoter 156341.doc -40 - 201201819 The efficient transcriptional activation of the gene" shows RNAa in human cells using synthetic dsRNA (called "small live RNA" (saRNA). It is not yet known whether RNAa is conserved in other organisms. Small double-stranded RNA (dsRNA) (eg, small interfering RNA (siRNA) and microRNA (miRNA)) has been found to be a trigger for the evolutionary conserved mechanism of RNA interference (RNAi). RNAi is usually remodeled via chromatin. This represses transcription to cause gene silencing, thereby degrading complementary mRNA, or blocking protein translation. However, in the context of the detailed description of the Examples section below, it is shown that oligonucleotides can enhance the expression and/or function of a BCL2 11 (BCL2L11)-like polynucleotide and its encoded product. "dsRNA can also be used as a small activating RNA ( saRNA). Without wishing to be bound by theory, the sequence 'saRNA can be induced by the gene in the dry gene promoter to induce target gene expression, a phenomenon known as dsRNA-induced transcriptional activation (RNAa). In another embodiment, the "preferred target segment" identified herein can be used to screen for additional compounds that modulate the expression of a BCL2 11 (BCL2L11) polynucleotide. "Modulators" are the following compounds: Or increase the expression of a nucleic acid molecule encoding BCL2L11, and at least include a 5-nucleotide portion that is complementary to the preferred target segment. The screening method comprises the steps of contacting a preferred target segment of a nucleic acid molecule encoding a sense or natural antisense polynucleotide of BCL2L11 with one or more candidate modulators, and selecting one or more to reduce or increase the encoding of BCL2L11. Candidate modulators of the expression of nucleotide nucleic acid molecules (eg, SEq m NO: 4 to 13). If one or more candidate modulators are shown to be capable of modulating (eg, reducing or increasing) the performance of a nucleic acid molecule encoding a bCL2LU polynucleotide, the modulator can be used in BCL2L1丨 polynucleotide work 156341.doc • 41 201201819 Investigative studies, or as a research, diagnostic, or therapeutic agent of the invention. Targeting a natural antisense sequence preferably modulates the function of the target gene. For example, the BCL2L11 gene (for example, accession number is nm_207002). In one embodiment, the antisense polynuclear acid of the stem BCL2L11 gene is acid. In one embodiment, the antisense oligonucleotide targets a sense and/or natural antisense sequence of a BCL2L11 polynucleotide (eg, accession number NM-207002), variants thereof, alleles, isozymes Types, homologs, mutants, derivatives, fragments and complementary sequences. Preferably, the oligonucleotide is an antisense molecule and the target comprises an encoding and non-coding region of an antisense and/or sense BCL2L11 polynucleotide. Preferred target segments of the invention may also be combined with their corresponding complementary antisense compounds of the invention to form stable double-stranded (duplex) oligonucleotides. It has been shown in the art that these double-stranded oligonucleotide moieties can modulate target expression and regulate translation and RNA processing via antisense mechanisms. In addition, the double-stranded moiety can be chemically modified. For example, it has been shown that the double-stranded portions can inhibit the target by typical hybridization of the antisense strand of the duplex to the target, thereby triggering enzymatic degradation of the target. In one embodiment, the antisense raised nucleotide Targeted BCL2 11 (BCL2L11) polynucleotides (eg, accession number NM_207002), variants thereof, alleles, isoforms, homologs, mutants, derivatives, fragments, and complementary sequences. Preferably, the oligonucleotide is an antisense molecule. According to an embodiment of the invention, the target nucleic acid molecule is not limited to BCL2L11 but extends to any of the isoforms, receptors, homologs and the like of the BCL2L11 molecule. 156341.doc • 42-201201819 In one embodiment, the oligonucleotide targets a natural antisense sequence of a BCL2L11 polynucleotide (eg, the polynucleotides set forth in SEQ ID NOS: 2 and 3), and Any of its variants, alleles, homologs, mutants, derivative 'fragments and complementary sequences. Examples of antisense oligonucleotides are set forth in SEQ ID NOS: 4 to 13. In one embodiment, the oligonucleotide complements or binds to a nucleic acid sequence of a BCL2L11 antisense molecule (including, without limitation, a non-coding sense and/or antisense sequence associated with a BCL2L11 polynucleotide) and modulates a BCL2L11 molecule Performance and / or function. In one embodiment, the oligonucleotide complements or binds to the nucleic acid sequence of the BCL2L11 natural antisense molecule (as set forth in SEQ ID NOS: 2 and 3) and modulates the expression and/or function of the BCL2L11 molecule. In one embodiment, the raised nucleotide comprises the sequence of at least 5 contiguous nucleotides of SEQ ID NOs: 4 to 13 and modulates the expression and/or function of the BCL2L11 molecule. Polynucleotide targets include BCL2L11 (including family members thereof), variants of BCL2L11; mutants of BCL2L11, including SNPs; non-coding sequences of BCL2L11; alleles of BCL2L11; species variants, fragments, and the like. Preferably, the oligonucleotide is an antisense molecule. In one embodiment, the oligonucleotides that target the BCL2L11 polynucleotide include: antisense RNA, interfering RNA (RNAi), short interfering RNA (siRNA); micro interfering RNA (miRNA); small temporal RNA (stRNA) Or short hairpin RNA (shRNA); small RNA-induced gene activation (RNAa); or small activating RNA (saRNA). 156341.doc -43- 201201819 In one embodiment, the 'BCL2 11 (BCL2L11) polynucleotides (e.g., SEQ ID NOs: 2 to 13) can modulate the performance or function of such targets. In one embodiment, the performance or function is up-regulated compared to the control group. In one embodiment, the performance or function is downregulated as compared to a control group. In one embodiment, the antisense compound comprises the sequences set forth in SEQ ID NOs: 4 to 13. The sigma 4 oligos may include one or more modified nucleotides, shorter or longer fragments, modified linkages, and the like. In one embodiment, SEQ id NO: 4 to 13 comprise one or more LNA nucleotides. Table 1 shows exemplary antisense oligonucleotides for use in the methods of the invention. Table 1 ··

Sequence number antisense sequence name sequence SEQ ID NO: 4 CUR-1519 G*T*T*C*C*A*C*C*T*T*C*T*C*C* D *C*C*C *A*G SEQ ID NO: 5 CUR-1520 C*T*G*C*T*G*T*C*C*T*A*G*T*G*T*C*T*C3*eCi* eC SEQ ID NO: 6 CUR-1523 G*T*C*T*G*T*T*G*C*T*T*C*C*C* D *C*C*T*C*T SEQ ID NO: 7 CUR-1525 G*G*A*G*G*G*A*G*G*T*G*T*G*T*G*T*G*T*G*T SEQ ID NO: 8 CUR-1524 C*C*T*T*C*T*T*T*C*T*C3|:C:*:T*C*A*C*A!*tC*CsiEC SEQ ID NO: 9 CUR -1526 G*C*C*A*C*C*C*A*G*T*A*C*A*C*A*C*A*C*C*T SEQ ID NO: 10 CUR-1521 〇 *0*0*Α*Τ*Τ*0*0*Τ*Τ*Τ*0*0*Τ*0*0*ϋ*Τ*0*Α SEQ ID NO: 11 CUR-1522 g*c* t*g*c*t*a*c*t*c*t*c*a*c*c*c*a*c*a*t SEQ ID NO: 12 CUR-1174 C*C*A* 0ΦΤ *0* A*G*G* G*C*A* A*C*A* C*A*G*C*A SEQ ID NO: 13 CUR-1505 C*C*T*C*T*C* C*A*C*G*C*G*C*A*G*T*A*C*A*T*T The desired target nucleic acid can be modulated in a number of ways known in the art. For example, antisense oligonucleotides, siRNA, and the like are used. An enzymatic nucleic acid molecule (e.g., a 'ribosinase) is a nucleic acid molecule capable of catalyzing one or more different reactions, including 156341.doc • 44-201201819, capable of repeatedly cleaving other individual nucleic acid molecules in a nucleotide base sequence-specific manner. The enzymatic nucleic acid molecule can be used, for example, to target substantially any rNA transcript. Because of its sequence specificity, trans-cleavable enzymatic nucleic acid molecules can be used as therapeutic agents for human diseases, and enzymatic nucleic acid molecules can be designed to cleave specific rnA targets in the background of cellular RNA. This cleavage event causes the mRNA to lose functionality and remove protein expression from the RNA. In this way, the synthesis of proteins associated with disease states can be selectively inhibited. In general, an enzymatic nucleic acid having an RNA cleavage activity functions by first binding to a target RNA. The binding is carried out via a target binding portion of the enzymatic nucleic acid. The target binding portion is immediately adjacent to the enzymatic portion of the molecule for cleavage of the target RNA. Thus, the enzymatic nucleic acid first recognizes the target RNA and then binds to the target RNA via a complementary base pair' and acts in an enzymatic manner to cleave the target RNA upon binding to the exact site. This cleavage strategy for a target RNA will disrupt its ability to direct synthesis of the encoded protein. After the enzymatic nucleic acid has bound and cleaves its RNA target, it is released from the RNA to find another target and can repeatedly bind and cleave the new target. Several methods, such as in vitro selection (evolution) strategies (〇rgel, (1979) pr〇c 1 London' B 205, 435), have been used to generate various reactions (eg, phosphodiester linkages and guanamine linkages). New nucleic acid catalyst for cleavage and ligation). The development of ribozymes with optimal catalytic activity will significantly contribute to any strategy that uses RNA lytic ribozymes to regulate gene expression. For example, 5, hammerhead ribozyme is active at a catalytic rate (kcat) of about 1 min-l under saturation of a concentration of Mg2+ cofactor 156341.doc -45-201201819. Artificial "RNA ligase" has been shown. The ribozyme can catalyze the corresponding self-modification reaction at a rate of about 100 min-1. In addition, it is known that certain modified hammerhead ribozymes have a binding arm composed of DNA, which can be close to 100 min-1. The turnover rate catalyzes the cleavage of RN A. Finally, the use of certain nucleotide analogs in place of the specific residues in the hammerhead ribozyme catalyzed core produces a modified ribozyme whose catalytic rate is shown to increase by up to 10 fold. It is shown that the ribozyme can significantly increase the catalytic rate of the catalytic rate shown by the majority of the natural self-cleaving ribozyme in vitro to promote chemical conversion. The structure of some self-cleaving ribozymes can be optimized for maximum catalytic activity, or can be prepared. A novel RNA motif showing a significantly faster rate of RNA phosphodiester cleavage. The intermolecular cleavage of RNA receptors catalyzed by RNA catalysts in accordance with the "hammerhead" model was first shown in 1987 (Uhlenb) Eck, 0·C. (1987) Nature, 328: 596-600). The RNA catalyst is recovered and reacted with a plurality of RNA molecules to indicate that it is indeed catalytic. Based on the "hammerhead" motif design Catalytic RNA has been used to cleave specific target sequences by making appropriate base changes in the catalytic RNA to maintain pairing with the necessary bases of the sequence. This allows the use of catalytic RN A to cleave specific target sequences. And indicates that the catalytic RNA designed according to the "hammerhead" model can cleave specific receptor RNA in vivo. RNA interference (RNAi) has become an effective tool for regulating gene expression in mammalian and mammalian cells. The coding sequence for the expression plasmid or virus and the small hairpin RNA for processing into siRNA delivers small interfering RNA (siRNA) in the form of RNA itself or DNA. This system enables 156341.doc -46-201201819 enough to siRNA precursor Efficient delivery into its active cytoplasm and allows the use of regulated and tissue-specific promoters for gene expression. In one embodiment, the nucleotide or antisense compound includes ribonucleic acid (RNA) And/or an oligomer or polymer of deoxyribonucleic acid (DNA), or a mimetic, chimera, analog or homolog thereof, the term encompasses between naturally occurring nucleotides, sugars, and covalent nucleosides Oligonucleotides composed of (backbone) linkages, and oligonucleotides having non-naturally occurring portions and functioning in a similar manner. Such modified or substituted oligonucleotides are often superior to natural forms, Desirable properties such as enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases. An 'oligonucleotide or r antisense compound according to the invention comprises antisense oligonucleotides (eg RNA, DNA, mimetics, chimeras, analogs or homologs thereof), ribozymes, external leader sequences (EGS) Oligonucleotides, siRNA compounds, single- or double-stranded RNA interference (RNAi) compounds (eg, siRNA compounds), saRNAs, aRNAs, and other oligomeric compounds that hybridize to at least a portion of a target nucleic acid and modulate its function. Thus, it can be DNA, RNA, DNA-like, RNA-like, or a mixture thereof, or can be a mimetic of one or more of such substances. The compounds may be single chain, double chain, cyclic or hairpin type oligomeric compounds and may contain structural elements such as internal or terminal expansion, mismatch or loop. Antisense compounds are typically made in a linear form, but may be joined or otherwise formed into a cyclic and/or branched form. Antisense compounds can comprise, for example, constructs that are hybridized to form two strands of a fully or partially double-stranded compound or a single strand that has sufficient auto-complementarity to hybridize and 幵V to 7L king or a double-stranded compound. The two strands can be internally connected to 15634I.doc -47 - 201201819 to create a free 3' or 5, end or connectable to form a continuous hairpin type structure or loop hairpin type structure can have an overhang at the 51 or 3' end Extend single-chain features. The double-stranded compound may contain an overhang at the end as needed. Other modifications may include attachment to one end, a selected nucleotide position, a sugar position, or a coupling group attached to an internucleoside linkage. Alternatively, the two strands can be joined via a non-nucleic acid moiety or a linker group. In the form of a single-stranded strand, the dsRNA can be in the form of an auto-complementary hairpin-type molecule that folds in itself to form a duplex. Thus, the dsRNA can be fully or partially double stranded. Gene expression can be specifically regulated by stably expressing the dsRNA hairpin type in the transgenic cell line. When formed from a single strand of two strands, or in the form of an auto-complementary hairpin-type molecule (self-folding to form a duplex), the two strands (or regions in which the duplex forms a duplex) are Wats〇 The complementary RNA bond of the base pairing of the n_CHck method. After introduction into the system, the compounds of the invention may elicit one or more enzymes or structural proteins <action to effect cleavage or other modification of the target nucleic acid or may act via a mechanism based on a primary basis, in general, nucleic acid (including nucleus acid) ) can be described as "DNA-like" (ie, usually having one or more 2,_deoxy sugars and (usually) T rather than TJ bases) or "RNA-like" (ie, usually having one or more 2,-hydroxyl or 2,_modified sugars and (usually, the snails of the sulphonic and non-tau bases can be used as the structural type of the above species, most commonly type A and type B. It is believed that the two have a type B The structural oligoacids are "like dna" and they have a type A-like structure "RNA-like". In some (chimeric) examples, antisense compounds may contain A and 3_-type regions. The antisense compound of the month may comprise an antisense portion of about 5 to about 8 nucleotides 156341.doc • 48-201201819 (that is, about 5 to about 80 linked nucleosides), which means antisense The length of the antisense strand or portion of the compound. In other words, the single chain antisense compound of the invention comprises from 5 to about 80 cores. And the double-stranded antisense compound of the invention (eg, dsRNA) comprises a sense and antisense strand or portion of from 5 to about 80 nucleotides in length. Those skilled in the art will appreciate that the length of this coverage is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 '23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 '35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, Or an antisense portion of 80 nucleotides, or any range thereof. In one embodiment, the antisense compound of the invention has an antisense portion of from 1 Å to 5 nucleotides in length. It should be understood that the length of the antisense part is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 , 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 Nucleotide, or an oligonucleotide thereof in any range. In some embodiments, the oligonucleotide is 15 nucleotides in length. In one embodiment, the antisense or oligonucleotide of the invention The compound has an antisense portion of 12 or 13 to 30 nucleotides in length. Those skilled in the art will appreciate that the length of the antisense portion is 12, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 156341.doc • 49·201201819 nucleotides, or any range of antisense compounds thereof. In one embodiment, the oligomeric compounds of the invention also comprise variants in which different bases are present at one or more nucleotide positions in the compound. For example, if the first nucleotide is adenine, a variant containing thymidine, uridine or cytidine at this position can be produced. This can occur anywhere in the antisense or dsRNA compound. The compounds are then tested using the methods described herein to determine their ability to inhibit the performance of the target nucleic acid. In some embodiments, the homology, sequence identity or complementarity between the antisense compound and the target is from about 40% to about 60%. In some embodiments, homology, sequence identity or complementarity is from about 60% to about 70% «in some embodiments, 'homology, sequence identity or complementarity is from about 7% to about 80%. %. In some embodiments, homology, sequence identity or complementarity is from about 80% to about 90%. In some embodiments, the homology, sequence identity or complementarity is about 90%, about 92%, about 94%, about 95. /. , about 96%, about 97%, about 98%, about 99% or about 100%. In one embodiment, antisense oligonucleotides (eg, the nucleic acid molecules set forth in SEQ ID NOS: 4 to 13) include one or more substitutions or modifications. In one embodiment, the nucleotide is replaced by a locked nucleic acid (LNA). In one embodiment, the oligonucleotide targets one or more of the sense and/or antisense nucleic acid molecules of the coding and/or non-coding sequence associated with BCL2L11 and the sequences set forth in SEQ ID NOs: 1 to 3. Areas. The oligonucleotides also dried to the overlapping regions of SEQ ID NOS: 1 to 3. Certain preferred oligonucleotides of the invention are chimeric oligonucleotides. "Chimeric oligonucleotides" or "chimeras" in the context of the present invention are oligonucleotides comprising two or more 156341.doc • 50-201201819 chemically distinct regions, each region being at least A nuclear bitter acid composition. The oligonucleotides typically contain at least one modified nucleotide acid region that confers one or more beneficial properties (eg, increased nuclease resistance, increased cellular uptake, increased binding affinity to the target); and is capable of cleaving RNA : DNA or RNA: the region of the RNA heterozygote that is regulated by the enzyme. For example, RNase tethers can cleave the RNA endonuclease of the RNA:DNA duplex. Thus, 'activated RNase Η cleaves RNA stems, thereby greatly enhancing the efficiency of antisense regulation of gene expression. Thus, comparable results are generally obtained with shorter oligonucleotides when using chimeric cryptonucleotides compared to phosphorothioate deoxyoligonucleotides that hybridize to the same dry region. Detection of cleavage of an RNA target can generally be performed by gel electrophoresis and, if desired, related nucleic acid hybridization techniques known in the art. In one embodiment, the chimeric oligonucleotide comprises at least one modification to increase target binding affinity. The region, and (often) used as the region of RNAse Η. The affinity of the oligonucleotide to its target (in this case, the nucleic acid encoded), the temperature at which the Tm-based oligonucleotide dissociates from the target, is typically determined by measuring the Tm of the oligonucleotide/target pair, using spectrophotometry Method to detect dissociation. The higher the Tm, the greater the affinity of the oligonucleotide to the target. The chimeric antisense compounds of the invention may be formed as a composite structure of two or more oligonucleotides, modified nucleotides, oligonucleosides and/or nucleotide mimics as described above. Thus, compounds are also known in the art as hybrids or combinations. Representative U.S. patents that teach the preparation of such hybrid structures include, but are not limited to, U.S. Patent Nos. 5, 13,830, 5,149,797, 5,220,0,7, 5,256,775, 5,366,878, 156,341. .doc • 51-201201819 5,403,711, 5,491,133, 5,565,350, 5,623,065, 5,652,355, 5,652,356, and 5,700,922 each incorporated herein by reference. In one embodiment, the modified region of the oligonucleotide comprises at least one nucleotide modified at the 2' position of the sugar, preferably via 2,-decyl, 2'-indolyl-anthracene -Alkyl or 2'-fluoro modified nucleotide "In another embodiment, the rnA modification comprises a ribose for a pyrimidine, an abasic residue or a reverse base of the 3, end of the 2, fluorine, 2'-Amino and 2, anthracene-fluorenyl modification. These modifications are typically incorporated into the oligonucleotide and have been shown to have a higher Tm (i.e., higher dry binding affinity) for a given stem relative to the 2,-deoxyoligonucleotide. This increased affinity effect greatly enhances the inhibition of gene expression by RNAi oligonucleotides. RNAse Η cleavage of RNA: an endonuclease of an RNA strand in a DNA duplex; thus 'activation of this enzyme cleaves the RNA target, and thereby greatly enhances the efficiency of RNAi inhibition. Lysis of the rnA target can usually be visualized by gel electrophoresis. In one embodiment, the chimeric oligonucleotide is also modified to enhance nuclease resistance. The cells contain a variety of degradable nucleic acid exonuclease and endonuclease. A variety of nucleotide and nucleoside modifications have been shown to allow the oligonucleotides incorporated into the oxime to be more resistant to nuclease digestion than natural oligodeoxynucleotides. Nuclease resistance is typically achieved by oligos. The cells are incubated with the cell extract or isolated nuclease solution and are often measured over time by gel electrophoresis for the amount of remaining intact oligonucleotide. Oligonucleotides that have been modified to enhance their nuclease resistance remain intact for longer than unmodified oligonucleotides. A variety of oligonucleotides have been shown to enhance or confer nuclear resistance to S#. Currently 'oligonucleosides containing at least one thioglycolate modification 156341.doc • 52- 201201819 The acid is better. In some cases, oligonucleotides that enhance target binding affinity are also modified to independently enhance nuclease resistance. Specific examples of some preferred oligonucleotides to be used in the present invention include those including modified backbones, such as phosphorothioates, phosphotriesters, decyl phosphonates, Short-chain alkyl or cycloalkyl sugar linkages or short-chain heteroatoms or heterocyclic linkages* the best oligonucleotides with a phosphorothioate backbone and those with a heteroatom backbone The hetero atom backbone is especially CH2-NH--0--CH2, CH,--N(CH3)--0--CH2 [referred to as an anthranylene group (methylimido group) or MMI master Chain], (:112--0--:^(〇^3)--CH2, CH2 -N (CH3)--N (CH3)--CH2 and 0--N (CH3)--CH2 -CH2 The main chain in which the natural phosphodiester backbone is represented by 0--P--0--CH). The guanamine backbone disclosed by De Mesmaeker et al. (1995) Acc. Chem. Res. 28:366-374 is also preferred. Also preferred are oligonucleotides having a morpholino backbone structure (Summerton & Weeller, U.S. Patent No. 5,034,506). In another embodiment (eg, a peptide nucleic acid (PNA) backbone), the phosphodiester backbone of the oligonucleotide is replaced by a polyamine backbone, and the nucleotide is directly or indirectly bound to the nitrogen of the polyamine backbone Aza atom. Oligonucleotides can also include one or more substituted sugar moieties. Preferred oligonucleotides include one of the following groups at the 2' position: OH, SH, SCH3, F, OCN, OCH3 OCH3, OCH3 0(CH2)n CH3, 0(CH2)n NH2 or 0 (CH2 n) CH3 (n is from 1 to about 10); C1-C10 lower alkyl, alkoxy, substituted lower alkyl, alkaryl or aralkyl; Cl; Br; CN; CF3; OCF3; S--, or N-alkyl; 0--, S--, or N-alkenyl; SOCH3; S02 CH3; 0N02; N02; N3; NH2; heterocycloalkyl; heterocycloalkylaryl; Alkylamine; polyalkane 156341.doc •53-201201819 arylamino; substituted methoxyalkyl; RNA cleavage group; reporter group; immigration agent for improving the pharmacokinetics of oligonucleotides a group of properties, or a group for improving the pharmacodynamic properties of an oligonucleotide and other substituents having similar properties. A preferred modification comprises 2,-methoxyethoxy [2I-0-CH2 CH2 OCH3, also known as 2,-0-(2-methoxyethyl)]. Other preferred modifications include 2·-methoxy (2,-0-CH3), 2·-propoxy (2,_OCH2CH2CH3) and 2'-fluoro(π). # Other modifications can be made to other positions on the scorpion nucleus, especially 3, the 3, and 5, and 5, terminal nucleotides of the terminal nucleotide. The oligonucleotide may also have a sugar mimetic, for example, using cyclobutyl instead of pentofuran. Oligonucleotides may also be modified or substituted, in addition or alternatively, to include nucleobases (often abbreviated as "bases"). As used herein, "unmodified" or "natural" nucleotides include adenine (A), black mites, thymine (τ), cytosine (C), and uracil (U). Modified nucleotides include nucleotides found only in natural or nucleic acids, such as hypoxanthine, 6 methyl adenine, 5 to me pyrimidine, especially 5-methylcytosine (also known as 5-a Keto-2'-deoxycytosine and commonly referred to in the industry as 5_Me_c), % by cytosine (HMC), glycosyl HMC and gentiolysyl HMC; and synthetic nucleotides, for example, 2-amine Adenine, 2_(decylamino) adenine, oxazolyl) adenine, 2_(alkylalkylamino) adenine or other hetero-substituted alkyl adenine, 2-thiouracil, 2- Thiothymidine, 5-bromouracil, hydroxyindole uracil, 8-azaguanine, 7-deazaguanine, aminohexyl) glandular and 2,6-diamino-based. It may include the "common" test base in the industry (for example, muscle bitterness has been shown to replace 5_Me_C with nucleic acid: 156341.doc •54·201201819 Chain stability is increased by -(10) and is replaced by the current preferred test. Another modification of the oligocore (IV) involves chemically linking to the nutrient nucleotides or a plurality of conjugates that enhance the activity or cellular uptake of the oligonucleotides. These moieties include, but are not limited to, lipid moieties (eg, Cholesterol moiety), cholesteryl moiety, aliphatic bond (eg, dodecyl diol or - undecyl residue), polyamine or polyethylene glycol chain, or adamantane acetate. It is known in the art to include lipophilicity. Part of the oligonucleic acid, and the method of preparing the oligo-nucleic acid, for example, U.S. Patent Nos. 5,138,045, 5,218,1,5, and 5,459,255. Given all positions in the oligo(4) and There is no need for a modified modification, and in fact, the above-mentioned repairs may be included in a single scorpion nucleus or even included in the mono-nuclear acid in the nucleus acid. The present invention also includes The nucleotides of the defined chimeric oligonucleotides. In another embodiment, the nucleic acid molecule of the invention is conjugated to another moiety, including but not limited to abasic nucleotides, polyethers, polyamines, polyamines, peptides, carbohydrates, lipids, or poly Hydrocarbon compounds. Those skilled in the art will recognize that such molecules can be attached to one or more of any of the nucleic acid molecules at several positions on the sugar, base or phosphate group. The oligonucleotides used in the present invention can be prepared in a convenient conventional manner via well-known solid phase & formation techniques. Several suppliers, including Applied Biosystems, sell equipment for carrying out the synthesis. One other approach is well known to those skilled in the art of the current synthesis of oligonucleotides. It is also well known to use similar techniques to prepare other oligonucleotides such as phosphorothioate and alkylated derivatives 156341.doc-55-201201819. Similar techniques and commercially available modified DNA synthetic amidite and fixed pore glass (CPG) products (eg, DNA synthesis nuclei modified with biotin, luciferin, acridine or psoralen) are also well known in the art. Glucosidic acid and/or CPG (available from Glen Research, Sterling, VA) to synthesize fluorescently labeled, biotinylated or otherwise modified oligonucleotides, such as cholesterol modified oligonucleotides. According to the present invention, the use of modifications (for example, using LNA monomers) to enhance the efficacy, specificity and duration of action and broaden the administration route of oligonucleotides include chemical methods such as MOE, ANA, FANA, PS, etc. This is achieved by the use of LNA monomers in place of some of the monomers in the current oligonucleotide. The LNA modified oligonucleotides may have a size similar to the parent compound or may be larger or preferably smaller. Preferably, the LNA-modified oligonucleotide contains less than about 70%, more preferably less than about 60%, optimally less than about 50% of the LNA monomer, and is between about 5 and 25 nucleotides in size, More preferably between about 12 and 20 nucleotides. Preferred modified oligonucleotide backbones include, but are not limited to, phosphorothioates, palmitic phosphorothioates, dithiophosphates, phosphotriesters, aminoalkyl phosphates, decyl phosphonates, and Other alkyl phosphonates (including phosphonic acid 3, alkylene esters and palladium phosphonates), phosphinates, amino phosphates (including amino amino phosphates and amine amino phosphate groups) Alkyl esters), thiocarbonylamino phosphates, thiocarbonylalkyl phosphonates, thiocarbonylalkyl acid triesters, and borane phosphates having a common 3'-5' linkage, such backbones 2,-5, linked analogs, and those with opposite polarity (wherein adjacent nucleoside units are changed from 3'-5' to 5'-3' or from 2'-5' to 5' -2'). Also contains a variety of salts, mixed salts and free 156341.doc • 56- 201201819 acid form. Representative U.S. patents including the above-described dish bonding include, but are not limited to, U.S. Patent Nos. 3,687,808, 4, 9, 9,863, 4,476,301, 5, 23,243, 5,177,196, 5,188,897 , Nos. 5,264,423, 5,276,019, 5,278,302, 5,286,717, 5,321,131, 5,399,676, 5,405,939, 5,453,496, 5,455,233, 5,466,677, 5,476,925, 5,519,126 , 5, 536, 821, 5, 541, 306, 5, 550, 1 1 1 , 5, 563, 253, 5, 571, 799, 5, 587, 361, and 5, 625, 050, each of which is incorporated herein by reference. Preferred modified oligonucleotide backbones containing no phosphorus atoms have a backbone formed by short-chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatoms, and alkyl or cycloalkyl nucleosides Inter linkages, or one or more short chain heteroatoms or heterocyclic internucleoside linkages. The backbones include those having a morpholino linkage (partially formed from a nucleoside sugar moiety); a siloxane backbone; a sulfide, a sulfoxide, and a sulfone backbone; a formazan group and a thiopurine主 醯 主 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; Ruthenium-based backbone; pyruvate and continuous amine backbone; guanamine backbone; and other components with mixed N, 〇, s and CH2. Representative U.S. patents for oligonucleosides include, but are not limited to, U.S. Patent Nos. 5,034,506, 5,166,315, 5,185,444, 5,214,134, 5,216,141, 5,235'033, Nos. 5,264,562, 5,264,564, 5,4,5,938, 5,434,257, 156341.doc -57-201201819 5,466,677, 5,470,967, 5,489,677, 5,541,307, 5,561,225 , 5,596,086, 5,602,240, 5,610,289, 5,602,240, 5,608,046, 5,610,289, 5,618,704, 5,623,070, 5,663,312, 5,633,360, 5,677,437, and 5,677,439, Each is incorporated herein by reference. In other preferred oligonucleotide mimetics, both the sugar and the internucleoside linkage (i.e., the primary linkage) in the nucleotide unit are replaced by a new group. The base unit is retained to hybridize the compound to a suitable nucleic acid. One such oligomeric compound (an oligonucleotide mimetic that has been shown to have excellent hybridization properties) is referred to as a peptide nucleic acid (PNA). The sugar-backbone in the 'oligonucleotide in the PNA compound is replaced by a guanamine-containing backbone, specifically an aminoethylglycine backbone. The nucleotide is retained and bonded directly or indirectly to the aza nitrogen atom of the guanamine moiety in the backbone. Representative U.S. patents which teach the preparation of PNA compounds include, but are not limited to, U.S. Patent Nos. 5,539,082, 5,714,331, and 5,719,262 each incorporated herein by reference. Further teachings of PNA compounds can be found in Nielsen et al. (1991) Science 254, 1497-1500. An embodiment of the invention is an oligonucleotide having a phosphorothioate backbone and an oligonuclear having a hetero atom backbone, and in particular, -CH2-NH of the above-mentioned U.S. Patent No. 5,489,677 -0-CH2-, -CH2-N(CH3)-0-CH2-[referred to as methylene (mercaptoimine) or MMI backbone], -CH2-0-N(CH3)-CH2-, -CH2N(CH3)-N(CH3)CH2- and -0-N(CH3)-CH2-CH2-[where the natural phosphodiester backbone is represented as -QP-0-CH2-], and as mentioned above The amine backbone of U.S. Patent No. 5,602,240. Also preferred is 156341.doc -58 - 201201819 The preferred one is the nucleotide of the morpholino backbone structure of U.S. Patent No. 5,034,506. The modified oligonucleotide may also contain one or more substituted sugar moieties. Preferred oligonucleotides include one of the following in the 2' position: 〇H; F; 0-, S- or N-alkyl, 0-, S- or N-alkenyl; 0-, S- or N-alkynyl; or decyl-0-alkyl, wherein the alkyl, alkenyl and alkynyl groups may be substituted or unsubstituted C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl groups System Ο (CH2)n OmCH3, 0(CH2)n, OCH3, 0(CH2)nNH2, 0(CH2)nCH3, 0(CH2)n0NH2, and 0(CH2n0N(CH2)nCH3)2, where n and m It can be from 1 to about 10. Other preferred oligonucleotides include one of the following in the 2' position: C1 to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, 0-alkylaryl or 0-aryl Alkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, 0CF3, SOCH3, S02CH3, 0N02, N02, N3, NH2, heterocycloalkyl, heterocycloalkylaryl, aminoalkylamino, poly Alkylamino, substituted formyl group, RNA cleavage group, acceptor group, intercalator, pharmacokinetic properties of modified oligonucleotides, or pharmacodynamics of modified oligonucleotides a group of properties, and other substituents of similar nature. Preferred modifications include 2·-methoxyethoxyethoxy (2'-0-CH2CH20CH3, also known as 2'-0-(2-methoxyethyl) or 2'-MOE), ie, alkoxy Alkoxy group. Other preferred modifications include 2'-dimethylaminooxyethoxy (i.e., 0(CH2)20N(CH3)2 groups, also known as 2匕DMAOE, as described in the Examples below), And 2·-didecylaminoethoxyethoxy (also known in the art as 2'-0-didecylaminoethoxyethyl or 2·-DMAEOE, ie, 2'-0- CH2-0-CH2-N (CH2)2). 15634 丨.doc -59- 201201819 Other preferred modifications include 2·-methoxy (2,-〇CH3), 2'-aminopropoxy (2'-〇CH2CH2CH2NH2) and 2'-fluoro (2' -F). Similar modifications can be made at other positions on the oligonucleotide, especially 3, terminal nucleotides or 2,_5, linking the 3' and 5's of the sugar in the oligonucleotide, 5 of the terminal nucleotide Bit. The oligonucleotide may also have a sugar mimetic ', for example, a cyclobutyl moiety is used instead of the pentofuranosyl sugar. Representative U.S. patents which teach the preparation of such modified sugar structures include, but are not limited to, U.S. Patent Nos. 4,981,957, 5,1 18,800, 5,319,080, 5,359,044, 5,393,878, 5,446,137, 5,466,786, 5,514,785, 5,519,134, 5,567,81 1 , 5,576,427, 5,591,722, 5,597,909, 5,610,300, 5,627,053, 5,639,873, 5,646,265, Nos. 5,658,873, 5,670,633, and 5,700,920 each incorporated herein by reference. Oligonucleotides may also include nucleobases (often abbreviated as "bases" in the art) for modification or substitution. As used herein, "unmodified" or "natural" nucleotides include purines (adenine (A) and guanine (G)) and pyrimidine (T), cytosine (〇 and Uracil (u)). Modified nucleotides include other synthetic and natural nucleotides such as 5-methylcytosine (5-me-C), 5-hydroxydecyl cytosine, astragalus, hypoxanthine , 2_Amino adenine, adenine and guanine 6-mercapto and other alkyl derivatives, adenine and guanine 2_propyl and other alkyl derivatives, 2-thiouracil, 2-thiothymidine and 2-thiocytosine, 5-norform uracil and cytosine, 5-propynyl uracil and cytosine, 6-azouracil, cytosine and thymine, 5- Urine, bite (pseudo-urine), 4-thiourea cancer bite, 8-dentate, 8-amino, 8-thiol, 8-156341.doc •60· 201201819 thioalkyl, 8- Hydroxyl and other 8-substituted adenines and black mites, 5-halogenated, especially 5-bromine, 5-trifluoromethyl and other 5-substituted urethanes and cyanosines, 7-mercapto guanines and 7 - thiol adenine, 8-azaindene and 8-azadenine, 7-Deazaguanine and 7-deaza adenine and 3-azepine guanine and 3-deaza adenine. In addition, the nucleotides include those disclosed in U.S. Patent No. 3,687,808, the disclosure of which is incorporated herein by reference. 'The Concise Encyclopedia of Polymer

Science And Engineering', pp. 858-859, Kroschwitz, JI Editor' John Wiley & Sons, 1990, by Englisch et al., "Angewandle Chemie, International Edition", 1991, 30, p. 613 And their disclosure by Sanghvi, YS, Chapter 15, "Antisense Research and Applications", pp. 289-302, Crooke, ST. and Lebleu, Β ea" CRC Press, 1993. Some of these nucleotides In particular, it is used to increase the binding affinity of the oligomeric compound of the present invention. The δ haizhuo nuclear acid includes a 5-substituting "• 密 bite, 6-aza 〇 及 及 and n_2, Ν·6 and 0-6 substituted 嘌呤, including 2_Aminopropyl adenine, 5 propynyl uracil and 5-propynyl cytosine. It has been shown that methylcytosine substitution can increase the stability of the nucleic acid duplex 〇.6-l_2 °C (SanghVi, YS, Crooke, ST and Lebleu, ed., ed., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are currently preferred base substitutions, even more particularly with 2, 〇曱 〇曱 乙基 乙基 糖 修饰 修饰 修饰 修饰Representative U.S. patents for the preparation of the above modified nucleotides and other modified nucleotides include, but are not limited to, U.S. Patent No. 3,687, No. 15,634, doc-61 - 201201819, and No. 4,845,205, No. 5,130,302 , 5, 134, 066, 5, 175, 273, 5, 367, 066, 5, 432, 272, 5, 457, 187, 5, 459, 255, 5, 484, 908, 5, 502, 177, 5, 525, 71 1 and 5, 552, 540, Nos. 5,587, 469, 5, 596, 091, 5, 614, 617, 5, 750, 692, and 5, 681, 941, each of which is incorporated herein by reference. A portion or conjugate of the activity, cell distribution or cellular uptake of the oligonucleotide is chemically linked to the oligonucleotide. Such moieties include, but are not limited to, lipid moieties (eg, cholesterol moieties), cholic acid, thioethers (eg, , hexyl-S-triphenylmercaptothiol), thiocholesterol, aliphatic chain (eg, dodecanediol or undecyl residue), phospholipid (eg, di-hexadecyl-) Racemic-glycerol or triethylammonium 1,2-di-O-hexadecyl-racemic-glycerol-3-H-phosphonate), polyamine or polyethylene glycol chain, or adamantane Acetic acid, palmitoyl moiety, or octadecylamine or hexylamino-carbonyl-oxycholesterol moiety. Representative U.S. patents that teach the preparation of such oligonucleotide conjugates include, but are not limited to, U.S. Patent Nos. 4,828,979, 4,948,882, 5,218,105, 5,525,465, 5,541,313, 5,545,730. , 5,552,538, 5,578,717, 5,580,731, 5,580,731, 5,591,584, 5,109,124, 5,118,802, 5,138,045, 5,414,077, 5,486,603, 5,512,439, 5,578,718, 5,608,046, 4,587,044, 4,605,735, 4,667,025, 4,762,779, 156341.doc-62-201201819 4,789,737, 4,824,941, 4,835,263, 4,876,335, Nos. 4,904,582, 4,958,013, 5,082,830, 5,112,963, 5,214,136, 5,082,830, 5,112,963, 5,214,136, 5,245,022, 5, 254, 469, 5, 258, 506 No. 5,262,536, 5,272,250, 5,292,873, 5,317,098, 5,371,241, 5,391,723 No. 5,416,203, 5,451,463, 5,510,475, 5,512,667, 5,514,785, 5,565,552, 5,567,810, 5,574,142, 5,585,481, 5,587,371, 5,595,726, Nos. 5,597,696, 5, 599, 923, 5, 599, 928, and 5,688, 941 each incorporated herein by reference. Drug Research: The compounds of the invention may also be added to the area of drug research and dry validation. The present invention encompasses the use of the compounds identified herein and preferred target segments for drug research, which attempts to elucidate the presence of a BCL2 11 (BCL2L11)-like polynucleotide with a disease state, phenotype, or condition. relationship. The methods comprise detecting or modulating a BCL2L11 polynucleoside, which comprises contacting a sample, tissue, cell, or organism with a compound of the invention to measure the nucleic acid or protein content of BCL2LU polyphosphate at a time after treatment. And/or related phenotypes or chemical endpoints, and visually measured values with untreated samples or otherwise in accordance with the invention

Effectiveness. The trial implementation of another compound treatment to determine the target used to or for the determination of a particular gene product, or phenotype, is 15634 丨.doc.63·201201819 Evaluation of gene expression up-regulation or inhibition: by direct detection The presence of a nucleic acid in a cell or organism to assess the transfer of nucleic acid into a host cell or organism. This test can be achieved by a number of methods known to the art. For example, the presence of an exogenous nucleic acid can be detected by Southern blotting or by a poly-enzyme chain reaction (p(3) technique to induce (4) anisotropic amplification and a primer of the (4) di-sequence of the nucleic acid (4). A method is used to measure the performance of exogenous nucleic acids. In the second column, Northern blot and reverse transcription PCR (rt_pcr) can be used to detect and quantify mRNA produced from exogenous nucleic acids. It can also measure enzyme activity or reporters. Protein activity to detect the expression of RNA by exogenous nucleic acids. For example, the antisense regulatory activity can be indirectly measured based on the decrease in the performance of the dry nucleic acid, thereby indicating that the exogenous nucleic acid is produced in the effector. Primers can be designed and used to amplify the coding region of the target gene. Initially, the control gene model 35 can be created using the highest expression coding region from each gene, but any-brown or non-coding regions can be used. Each of the control genes is assembled by inserting a reporter region into the reporter region and its p〇ly(A) s number. These plasmids will be generated in the upstream part of the gene with a reporter gene and at 3, mRNA having a potential RNAi target in the coding region. The effectiveness of individual antisense oligonucleotides is analyzed by modulating the reporter gene. The reporter gene used in the method of the invention comprises acetaminonate synthase (AHAS), Alkaline phosphatase (AP), β-galactosidase (LacZ), β-glucuronidase (GUS), chloramphenicol acetyltransferase (CAT), green fluorescent protein (GFp), red fluorescent Protein (RFp), yellow light-emitting protein (YFP), cyan fluorescent protein (CFP), horseradish peroxidase 156341.doc -64- 201201819 (HRP), luciferase (Luc), nopaline synthase ( NOS), octopine carnitine synthase (OCS), and derivatives thereof. A variety of selectable markers that confer resistance to the following substances: ampicillin, bleomycin, chloramphenicol ( Chloramphenicol), gentamycin., hygromycin, kanamycin, lincomycin, methotrexate, phosphinothricin, ° puromycin, and tetracycline. Determination of the regulation of reporter genes Methods are well known in the art and include, but are not limited to, fluorescent methods (eg, fluorescence spectroscopy, fluorescence activated cell sorting (FACS), fluorescence microscopy), antibiotic resistance assays. The methods known elsewhere in the art and described elsewhere herein analyze BCL2L11 protein and mRNA expression. For example, immunoassays such as ELISA can be used to measure protein content. The BCL2L11 ELISA assay kit is commercially available, for example, from R&D Systems (Minneapolis, MN). In the examples, BCL2L11 expression in a sample treated with the antisense oligonucleotide of the present invention (for example, cells or tissues in vivo or in vitro) is evaluated by comparison with BCL2L11 expression in a control sample. (eg, mRNA or protein). For example, methods known to those skilled in the art can be used to compare the performance of a protein or nucleic acid to a simulated or untreated sample. Alternatively, the treated sample can be compared to a control antisense oligonucleotide (e.g., with altered or different sequences) depending on the desired information. In another embodiment, the difference between the performance of the treated sample and the different nucleic acids in the untreated test 156341.doc-65-201201819 (including any criteria that the investigator deems appropriate, eg, housekeeping genes) can be used to compare The difference between the performance of the BCL2L11 protein or nucleic acid in the treated sample and the untreated sample. The observed difference can be expressed, for example, in ratio or fractional form for comparison with the money rib (iv). In the actual sample, the content of BCL2LH mRNA or protein f in the sample treated with the antisense oligonucleotide of the present invention is increased or decreased by about 1.25 times to about the untreated sample or the sample treated with the control nucleic acid. 1 () times or higher. In an embodiment, the content of the coffee " claw or protein is increased or decreased by at least about 125 times, at least about 13 times, at least, 1.4 times, at least about! 5 times, at least about 6 times at least about! 7 times at least about 1.8 times, at least about 2 times, at least about 25 times, at least about 3 times, at least about 3.5 times, at least about 4 times, at least about 45 times, at least about 5 times, at least about 5-5 L, At least about 6 times, at least about 6.5 times, at least about 7 times, at least about 75 times, at least about 8 times, at least about 8.5 times, at least about 9 times, at least about 9.5 times, or at least about 10 times or more . Kits, Research Reagents, Diagnosis, and Therapy The compounds described herein are useful for diagnosis, treatment, and prevention, and can be used as kit research reagents and components. In addition, those skilled in the art typically employ antisense oligonucleotides that are capable of inhibiting gene expression with strong specificity to elucidate the function of a particular gene or to distinguish the function of individual members of a biological pathway. For use in kits and diagnostics and in a variety of biological systems, the compounds of the invention (alone or in combination with other compounds or therapeutic agents) can be used as a tool in differential and/or combinatorial analysis to interpret cells and tissues. A pattern of expression of a portion of a gene or the entire complement. 156341.doc •66-201201819 The term “biological system” or “system” as used herein is defined as any organism, cell, cell culture or tissue that exhibits, or is sufficient to represent, a BCL2 11 (BCL2L11) gene product. Such systems include, but are not limited to, humans, transgenic animals, cells, cell cultures, tissues, xenografts, grafts, and combinations thereof. - according to a non-limiting example, the expression pattern in a cell or tissue treated with one or more antisense compounds is compared to a control cell or tissue not treated with an antisense compound, and the resulting pattern is analyzed, for example, for disease-related Degree of difference in gene expression, sex pathway, cell localization, degree of expression, size, structure or function of the gene being tested. Such assays can be performed in stimulated or unstimulated cells and in the presence or absence of other compounds that affect performance patterns. Examples of methods for gene expression analysis known in the art include Dna arrays or microarrays, SAGE (serial analysis of gene expression), READS (restriction enzyme amplification of digestive enzymes), T0GA (general gene expression analysis), protein arrays and Proteomics, sequenced display (EST) sequencing, subtractive RNA fingerprinting (SuRF), subtractive selection, differential display (DD), comparative genomic hybridization 'FISH (fluorescence in situ hybridization) and mass spectrometry. The compounds of the invention are useful in research and diagnostics because such compounds can hybridize to nucleic acids encoding BCL2 11 (BCL2L11). For example, oligonucleotides that are not hybridized in efficiency and conditions useful as potent BCL2L11 modulators as disclosed herein are effective primers or probes under conditions that facilitate gene amplification or detection, respectively. Such primers and probes can be used in methods requiring specific detection of nucleic acid molecules encoding BCL2L11, and amplification of such nucleic acid fragments 156341.doc • 67-201201819 sub- for detection or for further study of BCL2L11. The antisense oligonucleotides (especially primers and probes) of the invention and the parent of the nucleic acid encoding BCL2L11 can be detected by means known in the art. Such means may include coupling the enzyme to the oligonucleotide, radiolabeled oligonucleotide, or any other suitable means of detection. Kits can also be made using the BCL2L11 content of these test samples. Those skilled in the art also utilize the specificity and sensitivity of antisense compounds in therapeutic applications. Antisense compounds have been used as therapeutic moieties to treat disease states of animals, including humans. Antisense oligonucleotide drugs have been safely and effectively administered to humans and many clinical trials are currently being performed. It is thus determined that the antisense compound can be a useful therapeutic modality that can be used in the treatment of cells, tissues, and animals, particularly humans. An animal (preferably a human) suspected of having a disease or condition which can be treated by modulating the expression of BCL2L11 polynucleotide is administered by administration of an antisense compound of the present invention at the time of /α therapy. For example, in one non-limiting embodiment, the methods comprise the step of administering to a subject in need of treatment a therapeutically effective amount of a BCL2L11 modulator. The BCL2L11 modulator of the present invention can effectively modulate the activity of BCL2L11 or modulate the expression of BCL2L11 protein. In one embodiment, the activity or performance of BCL2L11 in the animal was inhibited by about 10% compared to the control group. Preferably, the activity or performance of BCL2L11 in the animal is inhibited by about 30%. More preferably, the activity or performance of BCL2L11 in an animal is inhibited by 50% or more. Therefore, the oligomeric compound can modulate the expression of BCL2 11 (BCL2L11) mRNA by at least 10%, at least 50%, at least 25%, at least 30%, at least 40%, at least 50%, at least 156341, compared to the control group. Doc -68 · 201201819 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%. In one embodiment, the activity or performance of BCL2 11 (BCL2L11) in the animal is increased by about 10% compared to the control group. Preferably, the activity or performance of BCL2L11 in the animal is increased by about 30%. More preferably, the activity or performance of BCL2L11 in animals is increased by 50% or more. Thus, an oligomeric compound can modulate the performance of BCL2L11 mRNA by at least 10%, at least 50%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, compared to a control group. At least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%. For example, the degree of decline in the expression of BCL2 11 (BCL2L11) in serum, blood, adipose tissue, liver or any other body fluid, tissue or organ of an animal can be measured. Preferably, the cells contained in the fluid, tissue or organ analyzed comprise nucleic acid molecules encoding the BCL2L11 peptide and/or the BCL2L11 protein itself. The compound can be used in a pharmaceutical composition by adding a compound of the invention of effect 1 to a suitable pharmaceutically acceptable diluent or carrier. The uses and methods of the compounds of the invention may also be used for prophylactic purposes. Conjugates Another aspect of the oligonucleotides of the present invention involves the chemical attachment of one or more moieties or conjugates that enhance the activity, cell distribution or cellular uptake of the nucleus to the oligo(tetra) acid. The moieties or conjugates may comprise a coupling group that is bonded to a functional group such as a - or a hydrazine group. The coupling group of the invention 156341.doc-69·201201819 comprises a group of pharmacodynamic properties of an intercalating agent, a reporter molecule, a polyamine, a polyamine, a polyethylene glycol, a polyether, and a reinforcing oligomer. And groups that enhance the pharmacokinetic properties of the oligomer. Typical coupling groups include cholesterol, lipids, fats, biotin, sulfonate, folic acid, phenanthridine, anthraquinone, acridine, luciferin, rh〇damine, coumarin, and dyes. . In the context of the present invention, a group that enhances the pharmacodynamic properties comprises a group that improves uptake, enhances degradation resistance, and/or enhances sequence-specific hybridization to a target nucleic acid. In the context of the present invention, a group that enhances the pharmacokinetic properties of a drug comprises a group that improves the uptake, distribution, metabolism or secretion of a compound of the invention. A representative coupling group is disclosed in International Patent Application No. PCT/US92/09196, filed on Oct. 23, 1992, and U.S. Patent No. 6,287,860, the disclosure of But not limited to lipid fractions (such as cholesterol fractions), cholic acid, thioethers (eg, hexyl-5-trityl mercaptan), thiocholesterol, aliphatic bonds (eg, dodecadiol or eleven Base residue), phospholipid (for example, di-hexadecyl-racemic-glycerol or triethylammonium 1,2-di-O-hexadecyl-racemic-glycerol_3_H phosphonate) , a polyamine or polyethylene glycol chain, or adamantane acetic acid, a palmidine moiety, or a octadecylamine or a hexylamino-carbonyl-oxycholesterol moiety. The oligonucleotide of the invention may also be coupled to an active drug Substances, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fenbufen, ketoprofen' S)-(+)-praloprofen ((s)-(+)-pranoprofen), carprofen (carprofen), dansylsarcic acid (dansylsarc) Osine), 2,3,5-triiodophenylhydrazine 156341.doc •70· 201201819 acid, flufenamic acid, leucovorin, stupid and oxadiazine, chlorothiazide, diazepam, hydrazine Indomethicin, barbiturate, cephalosporin, a tranexamine, an antidiabetic, an antibacterial or an antibiotic. Representative U.S. patents which teach the preparation of such oligonucleotide conjugates include, but are not limited to, U.S. Patent Nos. 4,828,979, 4,948,882, 5,218,105, 5,525,465, 5,541,313, 5,545,730. , 5,552,538, 5,578,717, 5,580,731, 5,580,731, 5,591,584, 5,109,124, 5,118,802, 5,138,045, 5,414,077, 5,486,603, 5,512,439, 5,578,718, 5,608,046, 4,587,044, 4,605,735, 4,667,025, 4,762,779, 4,789,737, 4,824,941, 4,835,263, 4,876,335, 4,904,582, 4,958,013 , 5,082,830, 5,112,963, 5,214,136, 5,082,830, 5,112,963, 5,214,136, 5,245,022, 5,254,469, 5,258, injury, 5,262,536, Nos. 5, 272, 25G, 5, 292, 873, 5, 317, 098, 5, 371, 241, 5, 391, 723, 5, 416, 203, 5,451,463, 5,510,475, 5,512,667, 5,514,785, 5,565,552, 5,567,81G, 5,574,142, 5,585,481, 5,587,371, 5'595,726, 5,597,696 , Nos. 5,599,923, f 5,599,928 and 5,688,941. Formulations 156341.doc • 71· 201201819 The compounds of the invention may also be admixed with other molecules, molecular structures or mixtures of compounds (eg, liposomes, molecules directed to the recipient, oral, rectal, topical or other formulations), capsules Sealed, coupled or otherwise combined to aid in ingestion, distribution and/or absorption. Representative U.S. patents which teach the preparation of such formulations which facilitate ingestion, distribution and/or absorption include, but are not limited to, U.S. Patent Nos. 5,108,921, 5,354,844, 5,416,016, 5,459,127, 5,521,291, 5,543,165, 5,547,932, 5,583,020, 5,591,721, 4,426,330, 4,534,899, 5,013,556, 5,108,921, 5,213,804, 5,227, 170, 5, 264, 221, 5, 356, 633, 5, 395, 619, 5, 416, 016, 5, 417, 978, 5, 462, 854, 5, 469, 854, 5, 512, 295, 5, 527, 528, 5, 534, 259, 5, 543, 152, 5,556,948, 5,580,575, and 5,595,756 each incorporated herein by reference. Although it is not necessary to administer an antisense oligonucleotide in a vector context to modulate target expression and/or function, embodiments of the invention pertain to expression vector constructs for expressing antisense oligonucleotides, including promoters , a hybrid promoter gene sequence and having a strong constitutive promoter activity, or a promoter activity that can be induced under the desired conditions. In one embodiment, the practice of the invention involves the administration of at least one of the above antisense oligonucleotides using a suitable nucleic acid delivery system. In one embodiment, the system comprises a non-viral vector operably linked to a polynucleotide. Examples of such non-viral vectors include only oligonucleotides (e.g., any one or more of SEQ ID NOs: 4 to 156341. doc • 72 to 201201819 13) or combinations with suitable protein, polysaccharide or lipid formulations. . Additionally, suitable nucleic acid delivery systems comprise a viral vector, typically from at least one of an adenovirus, an adeno-associated virus (AAV), a helper cell-dependent adenovirus, a retrovirus, or a Japanese liposome-coagulated virus (HVJ) complex. The sequence of one. Preferably, the viral vector comprises a strong eukaryotic promoter (e.g., a cytomegalovirus (CMV) promoter) operably linked to a polynucleoside &

In addition, preferred vectors comprise viral vectors, fusion proteins and chemical conjugates. The retroviral vector comprises "(1) Xianqiao mouse leukemia virus and Hiy-based virus. A preferred HIV-based viral vector comprises at least two vectors, wherein the gag and P〇1 genes are from the HIV genome and the env gene is from another virus. DNA virus Preferably, the vector comprises a pGx vector (e.g., orthopoxvirus or avian pox mother vector), a herpesvirus vector (e.g., herpes simplex virus (HS v) vector), an adenoviral vector, and an adeno-associated viral vector. A compound of the invention encompasses a salt of any pharmaceutically acceptable salt, ester, or oxime, or is capable of providing (directly or indirectly) a biologically active metabolite or residue thereof after administration to an animal, including a human. Any other compound. The term "pharmaceutically acceptable salt" means that the compound of the present invention is physiologically and pharmaceutically acceptable, that is, it retains the desired biological properties of the parent compound and does not impart undesirable toxicology. The salt of the effect. For the oligocore. A preferred embodiment of a pharmaceutically acceptable salt and its use is further described in U.S. Patent No. 6,287, the disclosure of which is incorporated herein by reference. 156341 .d〇c • 73- 201201819 The present invention also encompasses pharmaceutical compositions and formulations containing the antisense compounds of the present invention. The pharmaceutical compositions of the present invention can be administered in a variety of ways, depending on the desired local or systemic treatment and the area to be treated. Administration may be local (including eye: administration and administration (4), including vaginal and rectal delivery), lung (eg by inhalation or spraying of powder or aerosol 'contained by nebulizer); intratracheal, intranasal , epidermis and percutaneous, oral or parenteral administration. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial (e.g., intrathecal or intraventricular) administration. For treatment of tissue in the central nervous system, administration can be by, for example, injection or infusion into the cerebrospinal fluid. The administration of antisense RNA in cerebrospinal fluid is described, for example, in "Methods for slowing famiiiai Als disease progression", U.S. Patent Application Publication No. 2007/0117772, the entire contents of which is incorporated herein by reference. When it is intended to administer an antisense oligonucleotide of the present invention to a cell of the central nervous system, one or more agents capable of promoting penetration of the subject antisense oligonucleotide through the blood-brain barrier may be administered. Injections can be made, for example, in the entorhinal cortex or hippocampus. The delivery of a neurotrophic factor by administering an adenoviral vector to a motor neuron in muscle tissue is described, for example, in "Adenoviral-vector-mediated gene transfer into medullary motor neurons", US Patent No. 6,632,427 The citations are incorporated herein by reference. It is known in the art to deliver a carrier directly to the brain (e.g., striatum, thalamus, hippocampus, or substantia nigra) and is described in, for example, "Adenovirus vectors for the transfer of foreign genes into cells", U.S. Patent No. 6,756,523. Of the central nervous system particularly in brain" 156341 .doc -74- 201201819 I-cited in this article. Administration can be by rapid administration by injection or by slow infusion or administration of a sustained release formulation over time. The antisense oligo (t) can also be used with drugs that provide the desired pharmaceutical or pharmacodynamic properties. For example, antisense (4) can be used in the industry to promote the penetration or transport of blood in the blood-brain barrier (such as iron). The antibody that transmits the protein stature is coupled and administered by intravenous injection. The antisense compound can be linked to a viral vector which, for example, can render the antisense a more efficient and/or increase the delivery of the antisense compound in the blood brain barrier. The osmotic blood-brain barrier can also be destroyed by, for example, infusion, including but not limited to meso-erythritol, xylitol, d(+), sugar, D(+) lactose, D (+) xylose, dulcitol, muscle_inositol, [(a) fructose, D(-) mannitol, d(+) glucose, d(+) arabinose, d(a) arabinose, cellobiose, D(+ Maltose, D(+) raffinose, L(+) rhamnose, D(+) melibiose, bismuth (-) ribose, ribitol, 〇(+) arabitol, l(-) arabin Alcohol ' D (+) fucose, L (-) fucose, D (-) to threose, L (+) to threose, and L (-) to threose; or amino acid, including but Not limited to glutamine, lysine, arginine, aspartame, aspartic acid, cysteine, glutamic acid, glycine, histidine, leucine, guanine , phenylalanine, valine, serine, threonine, tyrosine, proline, and taurine. Methods and materials for enhancing blood-brain barrier penetration are described, for example, in "Method for the delivery of genetic material across the blood brain barrier", No. 6,294, 520, "Material for passage through the blood-brain". "Blocker" and "Parental Delivery 156341.doc -75-201201819 systems", the entire contents of which are incorporated herein by reference. The title antisense compound can be mixed, encapsulated, coupled or otherwise combined with other molecules, molecular structures or mixtures of compounds (eg, liposomes, molecules that target receptors, oral, rectal, topical or other formulations). To facilitate uptake, distribution, and/or absorption. For example, cationic lipids can be included in the formulation to facilitate oligo(4) uptake. The composition shown to promote the capture is LIPOFECTIN (available from GIBCO-BRL, Bethesda, MD). According to hydrazine, at least one 2'-oxime-methoxyethyl modified oligonucleotide is particularly useful for oral administration. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Pharmaceutical carriers 'aqueous, powder or oily bases, thickeners and the like may be required or desired. Covered condoms, gloves, and the like can also be used. The pharmaceutical formulations of the present invention which may be conveniently presented in unit dosage form can be prepared according to conventional techniques well known in the pharmaceutical industry. These techniques comprise the step of bringing the active ingredient into association with a vehicle carrier or excipient. In general, the formulations may be prepared by uniformly and intimately bringing the active ingredient into the liquid carrier or the fine solid carrier or both, and then, if necessary, shaping the product. The compositions of the present invention may be formulated into any of a number of possible dosage forms such as, but not limited to, lozenges, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. The aqueous suspension may further contain a substance which increases the viscosity of the suspension, and contains, for example, carboxymethylcellulose 156341.doc • 76· 201201819 vitamin sodium, sorbitol and/or dextran. The suspension may also contain a stabilizer. The pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, foams, and formulations containing liposomes. The pharmaceutical compositions and formulations of the present invention may include one or more permeation enhancer' carriers, excipients or other active or inert ingredients. An emulsion is usually a heterogeneous system in which a liquid is dispersed in another droplet form (the diameter often exceeds 0.1 μm). The emulsion may contain, in addition to the dispersed phase, an additional component, and an active drug which may be present as a solution in the aqueous phase, the oil phase or as a separate phase. One embodiment of the invention comprises a microemulsion. Emulsions and their use are well known in the art and are further described in U.S. Patent No. 6,287,860. Formulations of the invention comprise liposome formulations. The term "liposome" as used in the present invention means a vesicle composed of two hydrophilic lipids disposed in one or more spherical bilayers. Lipid system monolayer or multilamellar vesicles having a film formed from a lipophilic material and an aqueous internal structure containing the composition to be delivered. Positively charged liposomes are positively charged liposomes which are believed to interact with negatively charged DNA molecules to form stable complexes. Liposomes that are sensitive or negatively charged are believed to capture DNA rather than complex it. Both cationic and non-cationic liposomes can be used to deliver DNA to cells. Liposomes also contain "stable" liposomes, as used herein, to refer to liposomes comprising one or more specific lipids. In inclusion in liposomes, such specific lipids produce liposomes with enhanced cycle life relative to liposomes lacking such specific lipids. Examples of sterically stabilized liposomes 15634 丨.doc •77· 201201819 The lipid moiety that forms vesicles in their liposomes includes one or more glycolipids or is derived from one or more hydrophilic polymers (eg, polyethylene glycol (pEG) ) Part)). Liposomes and their use are further described in U.S. Patent No. 6,287,860. The pharmaceutical formulations and compositions of the present invention may also comprise a surfactant. The use of surfactants in pharmaceutical products, formulations and emulsions is well known in the art. Surfactants and their use are further described in U.S. Patent No. 6,287,860, incorporated herein by reference. In one embodiment, the present invention employs various permeation enhancers to effect efficient delivery of nucleic acids, particularly oligonucleotides. In addition to facilitating the diffusion of non-lipophilic drugs in the cell membrane, penetration enhancers also enhance the permeability of lipophilic drugs. Penetration enhancers can be classified into one of five broad categories: surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants. Penetration enhancers and their use are further described in U.S. Patent No. 6,287,860, incorporated herein by reference. Cookers I should be aware that the formulation is usually designed according to its intended use (ie, the route of administration). Preferred formulations for topical administration comprise a blend of the oligonucleotides of the invention and a topical delivery agent, for example, lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents, and Surfactant. Preferred lipids and liposomes comprise neutral lipids and liposomes (eg, dioleoyl phosphatidylcholine DOPE ethanolamine, dimyristoylphosphatidylcholine DMPC, distearyl phosphatidylcholine), Negative lipids and liposomes (eg dimyristylphospholipid glycerol DMPG) and cationic lipids and lipids 156341.doc •78· 201201819 plastids (eg diterpenoid tetraf-aminopropylpropyl D〇TAp and Diterpenoid-phospholipid oxime ethanolamine DOTMA). For topical or other administration, the oligonucleotides of the invention may be encapsulated in or form complexes (especially cationic liposomes) with the ruthenium. Alternatively, the 'oligonucleotide can be complexed with a lipid, especially a cationic lipid. Preferred fatty acids and vinegar, pharmaceutically acceptable salts thereof, and their use are further described in U.S. Patent No. 6,287,860. Compositions and formulations for sigma administration comprising powders or particles 'microparticulate non-granules, suspensions or solutions in water or non-aqueous medium, capsules, gel capsules, sachets, lozenges or micro-forms Lozenges. It may be desirable to use an enhancer, diluent, emulsifier, dispersing aid or binder. Preferred oral formulations are those in which the oligonucleotides of the invention are administered in combination with one or more penetration enhancers, surfactants and sonicating agents. Preferably, the surfactant comprises sucrose and/or its g- or salt, bile acid and/or its salt. The preferred bile acids/isolates and their applications are further described in U.S. Patent No. 6,287, the disclosure of which is incorporated herein by reference. Also preferred are combinations of penetration enhancers, for example, a combination of a fatty acid/salt and a bile acid/salt. The combination of Youjia ^ ^ and UDCA. Other permeation enhancers include polyoxyethylene 9 lauryl shout, polyoxymethylene _2 〇 录 基 基 。. The oligonucleotide of the present invention may be orally delivered in the form of particles comprising spray dried particles or complexed to form micro or non-rice particles. Oligonucleotide complexes and their use are further described in U.S. Patent No. 6,287, the disclosure of which is incorporated herein by reference. Compositions and formulations for use in non-, & enteral, intra- or intra-ventricular administration may be included in 156341.doc • 79-201201819 containing sterile aqueous solutions which may also contain buffers, diluents and other suitable additives, such Other suitable additives are, for example, but not limited to, penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers or excipients. Certain embodiments of the invention provide pharmaceutical compositions comprising one or more oligomeric compounds and one or more other chemotherapeutic agents that function by non-antisense mechanisms. Examples of such chemotherapeutic agents include, but are not limited to, cancer chemotherapeutic drugs, such as daunorubicin, daunomycin, dactinomycin, doxorubicin, and softness. Epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, cytosolic aragonucleoside Cytosine arabinoside), bischloroethyl-nitrosurea, busulfan, mitomycin C, actinomycin D, mithramycin, prednisolone Prednisone, hydroxyprogesterone, testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine, quinone Melamine (pentamethylmelamine), mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea, nitrogen mustard Ustards), melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine , 5-azacytidine, hydroxyurea, pentastatin 156341.doc -80- 201201819 (deoxycoformycin), 4-hydroxyperoxycyclic guanamine (4· hydroxyperoxycyclo- Phosphoramide), 5-fluorouracil (5-FU), 5-fluorodeoxyuridine (5-FUdR), guanamine pterin (MTX), colchicine (c〇ichicines), taxol, vincristine, vinblastine, etoposide (VP-16), trimetrexate, irinotecan, Topotecan (t〇potecan), gemcitabine, teniposide, cispiplatin and diethylstilbestrol (DES). When used with a compound of the invention, the chemotherapeutic agents can be used alone (e.g., 5_FIJ and oligo-acidic acid), sequentially (e.g., 5-FU and oligonucleotides, followed by MTX and subsequent recruitment after a certain period of time). (nucleotide), or in combination with one or more of these other chemotherapeutic agents (eg, 5-FU, MTX and oligonucleotides, or 5-FU, radiation therapy, and oligonucleotides) use anti-inflammatory drugs (including but Not limited to non-steroidal anti-inflammatory drugs and corticosteroids, as well as antiviral drugs, including but not limited to ribivirin, vidarabine, acyci〇vir and ganciclovir (ganciclovir)) can also be combined in the compositions of the invention. Combinations of antisense compounds and other non-antisense drugs are also within the scope of the invention. Two or more combinations of compounds may be used together or sequentially. In another related embodiment, the compositions of the present invention may contain one or more antisense compounds directed to the first nucleic acid (especially oligonucleic acid (tetra)) and/or additional dry anti-sense compounds to the second nucleic acid target. For example, the first target can be a BCL211-like (BCL2U1)m-sense sequence and the fourth can be a region from another nucleotide sequence. Alternatively, the compositions of the present invention may comprise 156341.doc 81 201201819 antisense compounds having different regions of two or more dry-like BCL2 11 (BCL2L11) nucleic acid stems. A number of examples of antisense compounds are disclosed herein and other examples may be selected from suitable compounds known in the art. Compounds of two or more combinations may be used together or sequentially. Dosing: According to L', those skilled in the art should be aware of the formulation of the therapeutic composition and its subsequent administration (administration). Administration depends on the severity of the condition to be treated = the degree of response and the duration of the treatment may last from several days to several months, or until a cure or amelioration of the disease is achieved. The optimal dosing regimen can be calculated by measuring the drug in the patient = accumulation. Those skilled in the art can readily determine the optimal dosage, method of administration, and rate of repetition. The optimal dose may vary depending on the relative efficacy of the individual oligo nuclei and can generally be estimated based on the EC50 found to be effective in in vitro and in vivo animal models. Generally, the dose is 0.01 _g to 1 剂量. g/kg body weight, and can be given once or more daily, weekly or yearly: owed, or even every 2 to years. Those skilled in the art can readily estimate the rate of repetition of administration based on the measured residence time and concentration of the drug in the body fluid or tissue. After successful treatment, it may be desirable to have the patient maintain the therapy to prevent recurrence of the disease state, wherein the oligonucleotide is administered at a maintenance dose between 0.01 ton / kg body weight to 1 〇〇 g / kg body weight' and daily dose Once with - or more times to every 2 () years. In an embodiment, the patient is treated with a dose of at least about 1 mg/kg body weight, at least about 2 mg/kg body weight, at least about 3 mg/kg body weight, at least about 4 mg/kg body weight, at least about 5 mg. /kg body weight, at least about 6 I56341.doc • 82· 201201819 body weight, at least about 7 mg/kg body weight, at least about 8 mg/kg body weight, at least about 9 mg/kg body weight, at least about 10 mg/kg body weight, at least about 15 mg/kg body weight, at least about 20 mg/kg body weight, at least about 25 mg/kg body weight, at least about 3 mg/kg body weight, at least about 35 mg/kg body weight, at least about 4 mg/kg body weight, at least about 45 mg/kg body weight, at least about 50 mg/kg body weight, at least about 6 mg/kg body weight, at least about 70 mg/kg body weight, at least about 8 mg/kg body weight, at least about 9〇11^/1^ body weight , or at least about 1〇〇111§/4 body weight. Some of the injectable doses of the antisense oligonucleotides are described, for example, in "Antisense modulation 〇f PTP1B expressi〇n", U.S. Patent No. 7,563,884, the disclosure of which is incorporated herein by reference. Although various embodiments of the present invention have been described above, it should be understood that the implementations of the present invention are not limited to the disclosed embodiments. The spirit or scope of the invention. Therefore, the breadth and scope of the present invention should not be limited by any of the above embodiments. All documents of Fengwen and all of them are incorporated herein by reference. All publications and patent documents cited in this application are hereby incorporated by reference in their entirety in the extent of the the the the the The Applicant does not recognize that any particular reference is the "= surgery" of its invention for each reference cited in this document. Examples of compositions and methods of the invention are illustrated in the following examples. EXAMPLES The following non-limiting examples illustrate the elements of the components shown to illustrate selected embodiments of the invention. The ratio changes and alternatives have been exemplified by those skilled in the art and are within the scope of the embodiments of the invention. Example 1: Design of an antisense nucleic acid molecule of the class BCL2 11 (BCL2L11) and an antisense oligonucleotide specific for the sense strand of f or BCL21A 1 polynucleotide, as described above, the term "has "Specific oligonucleotide" or "targeted oligonucleotide" refers to an oligonucleotide having the following sequence: (1) capable of forming a stable complex with a portion of a targeted gene, or丨丨) is capable of forming a stable duplex with a portion of the mRNA transcript of the targeted gene.

Promoting the selection of appropriate oligonucleotides by using a computer program (eg IDT AntiSense Design, IDT 01ig〇AnalyZer) that automatically identifies sub-sequences of 19-25 nucleotides in each given sequence, The isomeric sequence forms a hybrid with the target polynucleotide sequence and has a desired melting temperature (typically 50-60 Å, and does not form a self-dimer or other complex secondary structure. By using an auto-aligned nucleic acid sequence And indicate a computer program of consensus or homology regions to further facilitate the selection of appropriate oligonucleotides.

The program compares the obtained nucleic acid sequences by, for example, searching a database such as GenBank or by sequencing the PCR products. Comparison of nucleic acid sequences from a range of genes and intergenic regions in a given genome allows for the selection of nucleic acid sequences of the appropriate degree of specificity for the target gene. Such procedures allow for the selection of oligonucleotides that exhibit a high degree of complementarity to a target nucleic acid sequence in a given genome and a lower degree of complementarity to other nucleic acid sequences.孰 This technology should be recognized as _, and the gene area applicable to the present invention can be selected in a larger form. 156341.doc • 84 · 201201819 Antisense compounds, "specifically hybridizable" in the following cases: the binding of a compound to a target nucleic acid can interfere with the target nucleus to regulate function and / or activity 'and exist a degree of complementarity to avoid the antisense compound and the non-dry nucleic acid sequence under conditions that are expected to specifically bind (ie, under physiological conditions in the case of in vivo analysis or treatment of pure treatment, and in the case of in vitro analysis) Non-specific binding occurs under conditions under which the assay is performed. The hybridization f of the oligonucleotides described herein can be carried out by one or more in vitro assays known in the art. For example, the nature of the oligonucleotides described herein can be obtained by measuring the binding strength between the dry natural antisense molecule and the potential drug molecule by (4) melting curve analysis. Any of the established methods of measuring the strength of the intermolecular interaction (e.g., melting curve analysis) can be used to estimate the binding strength between the target natural antisense molecule and the potential drug molecule (molecule). Melting curve analysis measures the temperature at which a natural antisense molecule/molecular complex rapidly transitions from a double strand to a single stranded form. This temperature has been widely accepted as a reliable measure of the strength of the interaction between two molecules. Melt curve analysis can be performed using a copy of the CDNA of the actual native antisense RNA molecule or a synthetic DNA or RNA nucleotide corresponding to the molecular binding site. Multiple kits containing all necessary reagents to perform this assay (e.g., Applied Biosystems, MeltDoctor kit) can be used. Such kits comprise a suitable buffer solution containing one of a double stranded DNA (dsDNA) binding dye (e.g., ABI HRM dye, SYBR Green, SYT0, etc.). The nature of the dsDNA dye is that it emits almost no fluorescence in free form, but has high fluorescence when bound to dsDNA. 15634l.doc * 85 · 201201819 To perform the analysis, the cDNA or the corresponding nutrient tannin is mixed with the molecule at a concentration defined by the specific manufacturer's protocol. The mixture is heated to 9 5 to dissociate all of the preformed dsDNA complexes and then slowly cooled to room temperature or other lower temperature as defined by the kit manufacturer to anneal the DNA molecules. The newly formed composite was then slowly heated to 95 ° C while continuing to collect data on the amount of fluorescence produced in the reaction. Fluorescence intensity is inversely proportional to the amount of dsC)NA present in the reaction. Real-time PCR instruments compatible with the kit can be used (eg ABIis

StepOne Plus Real Time PCR System or lightTyper instrument, Roche Diagnostics, Lewes, UK) to collect data. Plot the negative derivative of fluorescence relative to temperature (_d(fluorescence)/dT) 'y pumping) and temperature (X-axis) by using appropriate software (eg lightTyper (R〇che) or SDS Dissociation Curve, ABI) Line to build a molten bee. Analyze the data to determine the temperature at which the dsDNA complex rapidly transitions to a single-stranded molecule. This temperature is called Tm and is proportional to the strength of the interaction between the two molecules. Typically, the Tm is above 40 °C. Example 2: Modulation of BCL2L11 Polynucleotide Treatment of MCF-7 cells with antisense oligonucleotides All antisense nucleus acids used in Example 2 were designed as described in Example 1. The manufacturer (IDT Corporation Coralville, IA) was instructed to manufacture the designed phosphorothioate-bonded oligonucleotides and to provide the designed thioate analogs shown in the table. The star symbol between the nucleotides indicates the presence of a thiolate bond. The oligonucleotides required for the experiments in Example 2 can be synthesized using any suitable state of the art method, such as the method used for IDT: the use of phosphorous ruthenium on solid supports (e.g., 5 micron fixed glass beads (CPG)) Amine monomer (common nuclear bit I56341.doc -86 - 201201819 acid' all active groups are protected by protecting groups, such as trityl on sugar, benzamidine on A and C, and g on N-2-isobutyl fluorenyl). The protecting group prevents undesired reactions from occurring during oligonucleotide synthesis. The protecting group is removed at the end of the synthesis process. The initial nucleotide is attached to the solid support via 3' carbon and synthesized in the direction of 31 to 5,. Add a new base to the growing oligonucleotide strand in the following four steps: 〇 remove the protecting group from the 5' oxygen of the immobilized nuclear acid using tri-acetic acid; 2) use the tetrazole to immobilize the nucleotide Coupling with the next nucleotide in the sequence; the reaction is continued via the tetrazolylphosphinamide intermediate; 3) washing to remove unreacted free nucleotides and reaction by-products and unreacted immobilization Nucleotide capping to prevent its participation in the next round of synthesis; by using acetic anhydride and N-methylimidazole to acetylate the free 5· group to achieve capping; 4) to stabilize the bond between nucleotides, Use moths and water (if you want to generate acid diester bonds), or Beaucage reagents (3H-1,2-benzodithiol 3-keto-i,i-dioxide) (if desired phosphorothioate) Key) to bismuth phosphorus. The chimeric backbone can be constructed by alternately using two oxidizing agents. The above 4-step cycle is repeated for each nucleotide in the sequence. When the complete sequence is synthesized, the oligonucleotide is dissociated from the solid support using ammonium hydroxide at a high temperature and deprotection is carried out. The protecting group is removed by washing with desalting and the remaining oligonucleotide is lyophilized. To carry out the experiment designed in Example 2, at 37 ° C and 5% (: 02 in growth medium (MEM/EBSS (Hycl〇ne catalog number: SH30024, or Mediatech catalog number: MT-10-010-CV) +10. /. FBS (Mediatech catalog number: MT35-011-CV) + penicillin (stictotin) / streptomycin (Mediatech catalog number: MT30-002-CI)) growing MCF from ATCC 7 cells (Catalog No. HTB-22). On the day before the experiment, 156341.doc -87 · 201201819 The cells were plated again in a 6-well plate at a density of 1.5 χ 105/ml and at 37 ° C and 5% C02 Incubate overnight. On the day of the experiment, replace the medium in the 6-well plate with fresh growth medium. Oligonucleotides delivered by the manufacturer in lyophilized form were diluted to 20 μL in deionized water without RNAse/DNAse. 2 μΐ This solution was incubated with 400 μΐ OptiMEM medium (Gibco catalog number: 31985-070) and 4 μL Lipofectamine 2000 (Invitrogen catalog number: 1 1668019) for 20 min at room temperature and then applied dropwise to a 6-well plate. There is one well of MCF-7 cells. Use 2 μΐ water instead of oligo A similar mixture of acid solutions was used as a mock transfection control group. After incubation for 3-18 h at 37 ° C and 5% CO 2 , the medium was changed to fresh growth medium. After adding antisense oligonucleotide for 48 h, the medium was removed. Add 600 ng of extracted RNA to the RNA extracted from the cells using the SV Total RNA Isolation System from Promega (Catalog No.: Z3105) or the RNeasy Total RNA Isolation Kit from Qiagen (Catalog No. 741 81) according to the manufacturer's instructions. The reverse transcription reaction was performed using a Verso cDNA kit from Thermo Scientific (Catalog No.: AB1453B) or a high capacity cDNA retroviral kit (Catalog No.: 4368813) as described in the manufacturer's protocol. Use of this reverse transcription reaction The cDNA was subjected to real-time PCR using ABI Taqman Gene Expression Mix (catalog number: 43 69510) and primers/probes designed by ABI (Applied Biosystems Taqman Gene Expression Assay: HsOO 197982_ml >

Applied Biosystems, Inc., Foster City CA) to monitor gene expression. The following PCR cycle was performed using the StepOne Plus Real-Time PCR Machine (Applied Biosystems) 156341.doc • 88 - 201201819: 2 min at TC, 10 min at 95 min, 40 cycles (maintained at 95) 15 seconds, 丄min at 6 〇t). Based on the difference between the 18S normalized dCt values between the treated and simulated transfected samples, the fold change in gene expression after treatment with antisense oligonucleotides was calculated. : Real-time PCR results showed that the content of BIM mRNA in MCF7 cells was significantly increased after 48 h treatment with the polymer designed as BIM antisense Hs.652337. Treatment of HepG2 cells with antisense nucleosides 2 All antisense nucleoside acids used were performed as described in Example 1. 6 indicates that the manufacturer (IDT, Coralville, IA) manufactured the designed phosphorothioate-bonded oligonucleotides and provided a table. The designed thio acid vinegar analog shown in 1. The internucleotide star symbol indicates the presence of a phosphorothioate linkage. The nucleotides required for the experiments in Example 2 can be used in any suitable state of the art. Method to synthesize, for example, the method used by IDT: Sub-filled amide monomers (common nucleotides of common nucleotides are commonly used on bulk carriers (eg 5 micron fixed glass beads (CPG)), all of which are protected by protecting groups, such as trityl, A on sugar And the phenylhydrazine group on C and the N-2-isobutyl fluorenyl group on the oxime.) The protecting group prevents undesired reaction during the synthesis of the nucleotides. The protective group is removed at the end of the synthesis process. , carbon attaches the initial nucleotide to the solid support and synthesizes in the 3' to 5' direction. Add new bases to the growing oligonucleotide strand in the following 4 steps: 1) Self-fixation using tri-acetic acid The 5' oxygen at the nucleotide removes the protecting group; 2) the tetrazole is used to couple the immobilized nucleotide to the next nucleotide in the sequence; the reaction is continued via the tetrazolylphosphinium intermediate 3) washing to remove unreacted free nuclear bitter 15634l.doc -89 - 201201819 acid and reaction by-products and capping the unreacted fixed oligonucleotide to prevent its participation in the next round of synthesis; by using acetic anhydride and N-methylimidazole acetylated to the free 5, hydroxyl to achieve end-capping; 4) is a stable core The bond between acids, using moths and water (right to produce acid-filled diacetate), or Beaucage reagent (3H-1,2_benzodithiol-3-one-1,1-dioxide) (if desired) Phosphorothioate linkage) to oxidize phosphorus. The chimeric backbone can be constructed by alternately using two oxidizing agents. The above 4-step cycle is repeated for each of the nucleotides in the sequence. When the complete sequence is synthesized, the oligonucleotide is cleaved from the solid support at a high temperature using ammonium hydroxide and deprotected. The protecting group is removed by washing with desalting and the remaining oligonucleotide is lyophilized. For the experiment designed in Example 2, the growth medium (MEM/EBSS (Hyclone catalog number: SH30024, or Mediatech catalog number: MT-10-010-CV) + 10% FBS at 37 ° C and 5% CO 2 ). HepG2 cells from ATCC (Catalog No.: HB-8065) were grown in (Mediatech catalog number: MT35-011-CV) + penicillin/streptomycin (Mediatech catalog number: MT30-002-CI)). On the day before the experiment, the cells were plated again in a 6-well plate at a density of X·5 X 1 〇 4/mi and incubated overnight at 37 ° C and 5% CO 2 . On the day of the experiment, the medium in the 6-well plate was changed to a fresh growth medium. The oligonucleotides delivered by the manufacturer in a cool dry form were diluted to a concentration of 20 μM in deionized water without RNAse/DNAse. 2 μL of this solution was incubated with 400 μΐ OptiMEM medium (Gibco catalog number: 3 1985-070) and 4 μL Lipofectamine 2000 (Invitrogen catalog number: 1 1668019) for 20 min at room temperature, then applied dropwise to 6 wells 156341 .doc -90- 201201819 The plate has one well of HepG2 cells. A similar mixture containing 2 μM water instead of the oligonucleotide solution was used as a mock transfection control group. After incubation for 3-18 h at 37 ° C and 5% CO 2 , the medium was changed to a fresh growth medium. After 48 h of antisense nucleotide addition, the medium was removed and the SV Total RNA Isolation System from Promega (catalog number: Z3105) or the RNeasy Total RNA Isolation kit from Qiagen (catalog number: 74181) was used according to the manufacturer's instructions. RNA is extracted from cells. 600 ng of extracted RNA was added to the reverse transcription reaction using the Verso cDNA kit from Thermo Scientific (catalog number: AB1453B) or the high capacity cDNA reverse transcription kit (catalog number: 4368813), as in the manufacturer's protocol. Said. The cDNA from this reverse transcription reaction was used by real-time PCR using ABI Taqman Gene Expression Mix (catalog number: 43695 10) and primers/probes designed by ABI (Applied Biosystems Taqman Gene Expression Assay: Hs00197982_ml (BCL2Lll), Applied Biosystems , Foster City CA) to monitor gene expression. The following PCR cycle was performed using a StepOne Plus real-time PCR machine (Applied Biosystems): 2 min at 50 °C, 10 min at 95 °C, 40 cycles (15 seconds at 95 °C, at 60 °C) Hold at °C for 1 min). Based on the difference between the 18S normalized dCt values between the treated and simulated transfected samples, the fold change in gene expression after treatment with antisense oligonucleotides was calculated. Results: Real-time PCR results showed that the design was BIM antisense Hs. After 48 h of oligomer treatment of 652337, the content of BIM mRNA in HepG2 cells was significantly increased (Fig. 2). Treatment of primary keratinocytes with antisense oligonucleotides 156341.doc -91 - 201201819 Growth in growth medium (keratinocyte growth medium, Lifeline catalog number: LM0004 + Lifeline growth factor 1030) at 37 ° C and 5% C02 Primary keratinocytes (from Lifeline Technologies). On the day before the experiment, cells were plated again in a 24-well collagen-coated plate (Beckton Dickinson BioCoat plate, catalog number: 35 6408) at approximately 5 x 10 OA4/ml (or approximately 1/3 dilution of 90% confluence). Incubate overnight at 37 ° C and 5% CO 2 . On the day of the experiment, the medium in the 24-well plate was changed to 1 ml of fresh growth medium. Oligonucleotides delivered by the manufacturer in lyophilized form were diluted to 20 μL in deionized water without RNAse/DNAse, 2 μM of this solution and 400 μM OptiMEM medium (Gibco Cat. No. 3 1985-070) and 4 μΐ TransIT®-LT1 transfection reagent (Minis catalog number: MIR 2300) was incubated for 20 min at room temperature and then applied dropwise to a well of a 6-well plate with HepG2 cells. A similar mixture containing 2 μM water instead of the oligonucleotide solution was used as a mock transfection control group. After administration, the plates were incubated overnight at 37 ° C, 5% CO 2 . 24 h after the addition of the antisense oligonucleotide, fresh growth medium was used instead of the medium and the administration was repeated as described above. At the second dosing for 24 h, RNA was extracted from the cells using the SV Total RNA Isolation System from Promega (Catalog No.: Z3 1 05) according to the manufacturer's instructions. A total of 600 ng of RNA was added to the reverse transcription reaction, which was performed using the high capacity cDN A kit from Applied Biosystem (catalog number: 43688 13) as described in the manufacturer's protocol. The gene expression from this reverse transcription reaction was monitored by real-time PCR using ABI Taqman Gene Expression Mix (Catalog No. 4369510) and ABI-designed primer/probe (Hs00197982_ml). 156341.doc -92- 201201819 The following PCR cycle was performed using a StepOne Plus real-time PCR machine (Applied Biosystems): 2 min at 50 °C, 10 min at 95 °C, 40 cycles (at 95 °C) Hold for 15 seconds and hold at 60 ° C for 1 min). The fold change in gene expression after treatment with antisense oligonucleotides was calculated based on the difference between the 18S normalized dCt values between the treated and mock transfected samples. Results: Real-time PCR results showed a significant increase in the amount of BIM mRNA in keratinocytes after 48 h treatment with the oligomer designed to BIM antisense Hs. 6523 37 (Fig. 3). Example 3: In vitro apoptosis assay The extent of apoptosis in cells was determined using the HT Titer TACS assay kit according to the manufacturer's protocol (Trevigen catalog number: 4822-96-K). Briefly, cells after 48 hours of oligonucleotide transfection were trypsinized, counted and plated again in 96-well plates at a density of 100,000 cells/well. The 96-well plates were then incubated overnight at 37 ° C, 5% CO 2 . The next morning, the medium was removed and the cells were fixed in a 3.7% buffered formaldehyde solution (Sigma-252549) rinsed with PBS and 100% sterol (Sigma-M1 775-1GA) and stored at 80 at +4 °C. % ethanol (Sigma-4935 1 1). For labeling DNA fragmentation, ethanol was removed, and the cells were washed with PBS and incubated with proteinase K solution for 15 minutes at room temperature, washed in water and PBS, and then incubated with TACS nuclease solution at 37 ° C for 10 minutes. After the incubation, the cells were washed with PBS, and a hydrogen peroxide solution (Sigma-MKBD1394) was added at room temperature for 5 minutes. The cells were then incubated with the lx TdT marker for 5 minutes at room temperature. The buffer was removed and the cells were then incubated with the labeling reaction mixture for one hour at 37 °C, and then lx TdT stop buffer was added via 5 minutes 156341.doc -93 - 201201819. After the labeling reaction was stopped, the cells were washed with PBS, incubated with Strep-HRP solution for 10 minutes at room temperature, washed with PBS/Tween and incubated with TACS sapphire in the dark for 30 minutes. The reaction was stopped by the addition of 0.2 N HCl (Fluka-343102) and the absorbance was read in a plate reader at 450 nm. 1_A549 cells were transfected with 20 nM CUR-1174 (p63 antisense oligonucleotide) and CUR-1522 (BIM antisense oligonucleotide) using LipofectamineTM 2000 transfection reagent (Invitrogen) (ATCC No.: CCL -185TM). Results: Apoptosis was significantly increased in A549 cells transfected with CUR-1174 and CUR-1522 (Fig. 4). 2. Hep G2 cells (ATCC catalog number: HB-8065) were transfected with CUR-1519, CUR-1521 and CUR-1522 using LipofectamineTM 2000 Transfection Reagent (Invitrogen) at 20 nM. Cell subsets were also transfected with 20 nM CUR-1 505 (inactive oligonucleotide) and LipofectamineTM 2000 and water (simulated transfection). Results: Apoptosis was significantly increased in HepG2 cells transfected with CUR-1522. Other oligonucleotides have no significant effect on apoptosis (Figures 5 and 6). Although the invention has been illustrated and described in accordance with one or more embodiments, those skilled in the art Equivalent changes and modifications can be made after the figure. In addition, although specific features of the invention may be disclosed in accordance with one of the various embodiments, the features may be combined with one or more other features of other embodiments, which may be possible for any given or particular application. It is desirable and advantageous. 156341.doc • 94-201201819 The Abstract of the Disclosure allows the reader to quickly ascertain the nature of the technical disclosure. The Abstract is submitted with the understanding that it is not intended to limit or limit the scope or meaning of the following claims. References 1. Vogelstein B, Kinzler KW. Cancer genes and the pathways they control. Nat Med. 2004; 10:789-799. 2. Hanahan D, Weinberg RA. The hallmarks of cancer. Cell. 2000; 100:57-70 » 3· Evan GI, Wyllie AH, Gilbert CS et al., Induction of apoptosis in fibroblasts by c-myc protein. Cell. 1992 69:119-128. 4. Askew DS, Ashmun RA, Simmons BC, Cleveland JL. Constitutive c-myc expression in IL-3-dependent myeloid cell line suppresses cycle arrest and accelerates apoptosis. Oncogene. 1991; 6:1915-1922. 5. Klefstrom J, Vastrik I, Saksela E, Valle J, Eilers M, Alitalo K. c-Myc induces cellular susceptibility to the cytotoxic action of TNF-alpha. EMBO J. 1994; 13:5442-5450. 6. Lutz W, Fulda S, Jeremias I, Debatin KM, Schwab M. MycN and IFNy cooperate in apoptosis of human neuroblastoma cells. Oncogene. 1998; 17: 339-346. 7. Fadeel B, Orrenius S. Apoptosis: a basic biological phenomenon with wide-ranging implications in human 156341.doc -95- 201201819 disease. J Intern Med. 2005; 258:479-517 〇8. Lowe SW, Cepero E, Evan G. Intrinsic tumour suppression. Nature. 2004; 432:307-315 ° 9. Kaufmann SH, Vaux DL. Alterations in the apoptotic machinery and their potential role in anticancer drug resistance· Oncogene. 2003; 22:7414-7430 ° 10 Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer. 2005; 5: 876-885 o 11. Fischer U, Schulze-Osthoff K. New approaches and therapeutics targeting apoptosis in disease. Pharmacol Rev. 2005; 57:187-215 〇12. Reed JC. Drug insight: cancer therapy strategies based on restoration of endogenous cell death mechanisms. Nat Clin Pract Oncol· 2006; 3:388-398 0 13. Reed JC, Pellecchia M. Apoptosis-based Therapies for hematologic malignancies. Blood. 2005; 106:408-418. 14. Decaudin D, Marzo I, Brenner C, Kroemer G. Mitochondria in chemotherapy-induced apoptosis: a prospective novel target of cancer therapy (Review). Int J Oncol. 1998; 12:141-152 〇 15. Filomenko R, Pr0votat L, C, et al, Caspase-10 involvement in cytotoxic drug-induced apoptosis of tumor cells. Oncogene. 2006; 25:7635-7645 ° 16. Fulda S, Meyer E, Friesen C, Susin SA, Kroemer G, 156341.doc -96- 201201819

Debatin KM. Cell type specific involvement of death receptor and mitochondrial pathways in drug-induced apoptosis. Oncogene. 2001; 20:1063-1075. 17. Lacour S, Hammann A, Wotawa A, Corcos L, Solary E, Dimanche-Boitrel MT. Anticancer agents sensitize tumor cells to tumor necrosis factor-related apoptosis-inducing ligand-mediated caspase-8 activation and apoptosis. Cancer Res. 2001 61:1645-1651. 18. Micheau O, Solary E, Hammann A, Dimanche-Boitrel MT. Fas ligand-independent, FADD-mediated activation of the Fas death pathway by anticancer drugs. J Biol Chem. 1999; 19; 274:7987-7992. 19. Mollinedo F, Gajate C. Microtubules, microtubule-interfering agents and apoptosis. Apoptosis. 2003; 8:413-450 ° 20. Park SJ, Wu CH, Gordon JD, Zhong X, Emami A, Safa AR. Taxol induces caspase -10-dependent apoptosis. J Biol Chem. 2004; 279: 51057-51067. 21. Perkins C, Kim CN, Fang G, Bhalla KN. Overexpression of Apaf-1 promotes apoptosis of untreated and paclitaxel-or etoposide-treated HL-60 cells. Cancer Res. 1998; 58:4561-4566 o 22. Fridman JS , Lowe SW. Control of apoptosis by p53. Oncogene. 2003; 22:9030-9040. 156341.doc -97- 201201819 23. Yu J, Zhang L. The transcriptional targets of p53 in apoptosis control. Biochem Biophys Res Commun. 2005; 331:851-858 ° 24. Ashkenazi A, Pai RC, Fong S et al. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest· 1999; 104:155-162 e 25. Lee JM, Bernstein, A. p53 mutations increase resistance to ionizing radiation. Proc Natl Acad Sci USA. 1993; 90:5742 -5746 〇 26. Bunz F, Hwang PM, Torrance C, et al., Disruption of p53 in human cancer cells alters the response to therapeutic agents. J Clin Invest. 1999; 104:263-269. 27. Ashkenazi A, Holland P, Eckhardt SG. Ligand-based targeting of apoptosis in cancer: the potential of the recombinant human apoptosis ligand 2/tumor necrosis factor-related apoptosis-inducing ligand (rhApo2L/TRAIL). J Clin Oncol. 2008; 26:3621-3630 = 28. Ashkenazi A. Targeting death and decoy receptors of the tumour-necrosis factor superfamily. Nat Rev Cancer. 2002; 2:420-430 ° 29. Odoux C, Albers A, Amoscato AA, Lotze MT, Wong MK. TRAIL, FasL and a blocking anti-DR5 antibody augment paclitaxel-induced apoptosis in human non-small-cell lung cancer. Int J Cancer. 2002; 97:458-465 0 -98- 156341.doc

201201819 30. Miihlethaler-Mottet A, Bourloud KB, Auderset K, Joseph JM, Gross N. Drug-mediated sensitization to TRAIL-induced apoptosis in caspase-8-complemented neuroblastoma cells proceeds via activation of intrinsic and extrinsic pathways and caspase-dependent cleavage Of XIAP, Bcl-xL and RIP. Oncogene. 2004; 23:5415-5425 o 31. Ravi R, Jain AJ, Schulick RD, et al., Elimination of hepatic metastases of colon cancer cells via p53-independent cross-talk between irinotecan and Apo2 ligand/TRAIL.

Cancer Res. 2004; 64:9105-9114 ° 32. Jansen B, Schlagbauer-Wadl H, Brown BD et al, Bcl-2 antisense therapy chemosensitizes human melanoma in SCID mice. Nat Med. 1998; 4: 232-234. 33. Klasa RJ, Gillum AM, Klem RE, Frankel SR. Oblimersen Bcl-2 antisense: facilitating apoptosis in anticancer treatment. Antisense Nucleic Acid Drug Dev. 2002; 12: 193-213. 34. Letai A. Pharmacological manipulation of Bcl-2 family members to control cell death. J Clin Invest. 2005; 115:2648- 2655 ° 35. Ashkenazi A. Directing cancer cells to self-destruct with pro-apoptotic receptor agonists. Nat Rev Drug Discov. 2008; 7:1001-1012 〇 [Simplified Schematic] Figure 1 is a graphical representation of the results of real-time PCR, shown in phosphorothioate oligos introduced using Lipofectamine 156341.doc • 99 · 201201819 2000 The fold change + standard deviation of BCL2L11 mRNA after treatment with HepG2 cells compared with the control group. Real-time PCR results showed that the BIM mRNA content in MCF-7 cells was significantly increased 48 h after treatment with two oligomers designed to be BIM antisense Hs.652337. The bars designated CUR-1521 and CUR-1522 correspond to the samples treated with SEQ ID NOS: 10 and 11, respectively. Figure 2 is a graphical representation of real-time PCR results showing fold change + standard deviation of BCL2L11 mRNA compared to control group after treatment of HepG2 cells with phosphorothioate oligonucleotides introduced using Lipofectamine 2000. Real-time PCR results showed a significant increase in BIM mRNA levels in HepG2 cells 48 h after treatment with an oligomer designed to BIM antisense Hs.652337. The bars denoted as CUR-1519, CUR-1520, CUR-1523, CUR-1525, CUR-1524, CUR-1526, CUR-1521 and CUR-1522 correspond to the tests treated with SEQ ID NOS: 4 to 13, respectively. kind. Figure 3 shows the fold change in BCL2L11 mRNA expression in primary keratinocytes treated with phosphorothioate oligonucleotides. CUR-1522 significantly upregulates BIM mRNA in keratinocyte LL. The bar indicated as CUR-1522 corresponds to the sample treated with SEQ ID NO: 11. Figure 4 is a graph showing the results of apoptosis assay showing the percentage increase in apoptosis compared to the control group in A 5 4 9 cells treated with thiolatate S, which was introduced using Lipofectamine 2000. The results of cell apoptosis showed that CUR- significantly increased apoptosis in A549 cells compared with the control group. The bars corresponding to CUR-1174 and CUR-1522 correspond to 15634 丨.doc -100- 201201819 samples treated with SEQ ID NOS: 11 and 12. Figure 5 shows the percentage increase in cell apoptosis in cells treated with phosphorothioate oligonucleotides. Compared with the control group, CUR-1522 significantly increased apoptosis in cells. The bars indicated as CUR-1519, CUR-1521, CUR-•1522 and CUR-1505 correspond to the samples treated with SEQ ID NOS: 4, 10, 11 and 13. Figure 6 shows the percentage increase in cell apoptosis in cells treated with phosphorothioate oligonucleotides. Compared with the control group, CUR-1522 significantly increased apoptosis in cells. The bars designated CUR-1519, CUR-1521, CUR-1522 and CUR-1505 correspond to the samples treated with SEQ ID NOS: 4, 10, 11 and 13. BRIEF DESCRIPTION OF THE SEQUENCE LISTING - SEQ ID ΝΟ:1: Homologous human BCL2 11 (apoptosis promoter) (BCL2L11), transcript variant 9, mRNA (NCBI accession number: NM_207002); SEQ ID NO: 2: native BCL2L11 antisense Sequence (Hs.652337); SEQ ID NO: 3: native BCL2L11 antisense sequence (Hs. 618461); SEQ ID NOs: 4 to 13: antisense oligonucleotide. * indicates a phosphorothioate bond. 15634l.doc -101 201201819 Sequence Listing <110> American Okokena Limited Company <120> Treatment of BCL2L11 Related Diseases by Inhibition of Natural Antisense Transcripts Against BCL2 11 (BCL2L11)

<130> BIW <140> 100117636 <141> 2011-05-19 <150> 61/346,252 <151> 2010-05-19 <160> 13 <170> Patentln version 3.5 <210&gt ; 1 <211> 722 <212> DNA <213> Homo sapiens <300><308> NM_207002.2 <309> 2〇Tl-02-06 <313> (1)..( 722) < 400 > 1 acttcgctcc gcgcagccgc ctggtctgca gtttgttgga gctctgcgtc cagcgccgct 60 gccgctgccg ccgccgccgc cgccgccgcc gccgccgccg ccgccgccac taccaccact 120 tgattcttgc agccaccctg cgaaccctgc cacactgcga tcgcatcatc gcggtattcg 180 gttcgctgcg ttcccgccgc caccgcctcg gcgccctttc ttggcccttg ttcccccaaa 240 tgtctgactc tgactctcgg actgagaaac gcaagaaaaa aagaccaaat ggcaaagcaa 300 ccttctgatg taagttctga gtgtgaccga gaaggtagac aattgcagcc tgcggagagg 360 cctccccagc Tcagacctgg ggcccctacc tccctacaga cagagccaca agacaggagc 420 ccagcaccca tgagttgtga caaatcaaca caaaccccaa gtcctccttg ccaggccttc 480 aaccactatc tcagtgcaat ggtagtcatc ctagaggata taggtgatct ttcactgtgc 540 tttggattta tatttactgg cttagatttg tatggccacc accatagtc a agatacagaa 600 caactcaacc acaaggattt ctcatgatac ctttttatag ccacagccac ctctctccct 660 cttccttgag cattttgtca tatggtcatt ggtgattaaa taaaatgtat tttaatatig 720 ac 722 < 210> 2 < 211 > 3026 < 212 > DNA < 213 > Homo sapiens < 400 > 2 ctagtcctgt tgaactcaga tgactgcagc cacaggagtg atcccagagt cttttgcact 60 gggaggagaa ggtggaacag gagacaccca cattctagtt ggtccctgct gtctccaaga 120 ggtggtgtac caggagtcct acagatgtga agctaggtta aaaccagttc tggggatgct 180 ttcaaatcaa agaggattta aaaatgtgac tcccagttgc atttccggag ccaagcagca 240 tcctggcttg ggggcctggt gtctaccaca cttctgcgca ttcttcctcc aagccacatc 300 tcctgagaat aaagcaagat gccattggca atgtctactc agaactactt gaatgactca 360 tcaatcaaca ggcttgaatg ctcttcttcc tctatgattc aacggtttga ttgactgaac 420 tgaaactaaa acccaaccct agtggtcatt ttagacttga gcagatacag tcagaatctc 480 156341-Sequence table.doc 201201819 aatcacatgt catcagcagg ccctccttgg cctccttatt gcagctgggc tcctgagcag ctctccctca ctcagggagg aaacgaacgg ctcctttcag tagggcagac gaaggctgct ggctgacgca actgctcctg tgtccacttc cca gcaaggg tgcaaaacag gatttgtgct tgtgctggtg aatctgggct tcacctagca aatcaggggg acaccaaaat gaaaacagcc ctagaggcat ggagaaagcc tcaggtgcac taa ^ gtgcca acagaaacga tgcatctcag agttgacagt catgactcaa ataggacgtg aaggcaacat gtgggtgaga gtagcagcta tgggtgtaaa tatgatcaaa latgggggag gtgtccagat ttcttggaca tggttacaat cagtatttat tttctctctt agtgaacgag tttttggttt ttcaatactg ctaattttac aggcaattca ctacgttccc tgggaggtgg agtggccttc ctccctgctg tgcgtgggtt acacagcctg cctcacttcc ctgtgggtcc ttcatcacct gcatgctcac atgattccat tatttgagct catggcagga aatagaacct gattcaacat tttgctaagt attattttca ccacactgaa tagggtcctt itttgatctg caacccacag ctgggctgtg gttctctcaa cacaggtaga ctgaaaagct tcctcctgca ttgatttctc agcatgggct gaccacatgt tcacaagtac ttgttctut ccatacgtcc caatgcaggc aggoaclggc agagcaggac agctgtgtcc aggagttcag ggaacaaaat aaggctactg taacttcagt caacttcaag gcacggtgta aaataaagta agagattatc agtcaaatag ccatttgcat taaaattctc cactttaaga agctgagatc tttgctttat tttttaaaaa gcaaaatgaa gtcagtttta atggagatat acaattgtta actgttgggt cattttgaaa ggc tttcttc cttaaaatga gtctcagctg atagctactc actaatgctg ataattatct gaagcaaaat aaatacaaat gtctgggtta atgaagactg aaacaggata atgcctggac actaaatttt ccaagaacaa ggaacacaat tgttctctac atacccctgc aaaaatgctt aatggccagg acacaggaat cttgtgtact tagagacctt gttaggatgg agccccgggg gcaccaccac tggccttcga gccaaggtct cacagagagg gcagcaaggg ggagcagctc atcctcctcc ccagctaggg agacactagg acagcagtgg agtggagtcc gaaatctgcc atagaagccc cagcttacta tttgctcact gagcagcaag tcaacctcag tttctctgtg cctacagggg agtaacagct glicacacta aagggtgctt gtgaggatga tataagaatg aatatggacc tgctctgaca acactgaagt tccagacaaa agaataggca ttagttatct gatttgaag £ actgtggggg attggaattt taaaaataaa cctcaaccca actccttctc ttgtggggcc tcggttgaac aactgggcac ttgcactgcc ttctgaatat gtggatggat tttgcctgct ttggagaagc atatgaactc ctcagggcat ctagtgccct agagtgtgcc aaaggaacaa ggaggactac aaatgggtga [gcctggagc ttaacccacc tccatUggg atlcggagct ctggtttctg tgttcagcia gaatcttcga cagtcattta atgtctctct ttcttagttt tactcatttg ttgaatgggg attatattag ccttactttc cttgaaggtc ttaatgagaa t gaaatgaga taatttttaa gtatatatat gcatttctaa tttccatcag actggagcgt cggtataagt cctgggtgtt ctggaaagtt ttatttattt agaaacagtt taggctgatt gctatctccc tatgctaact ttttgttttt ttagtaaact tutattttg gaatactttc aaatttacag aaaagtttca aagataacaa gagttctcat ttgcccttca cccagtttcc ccataactat 156341- Sequence Listing .doc • 2 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700

201201819 ggctcattgt aaaaactagg aaaccaatat taatgcatta ctaaiaacta tagactttat 2760 tcagatttta ccagtttttc tactcatgtt cttctgtccc aggattcaa Shu acatgatact 2820 gcattgtatt tagttaccat atctcctgag tctcttctgc tctgtgatgg cttcagtctt 2880 ttttcgtctt ccatgcatcg tcagttttaa tgggtactag gcaggtatta tggacaagct 2940 tggcaaatcc acagcccatg ggccacatga ggcccaggat aactttgaat gtggcccagt 3000 acaaattcat aaacttttta aaaaac 3026 < 210 > 3 < 211 > 1512 < 212 > < 213 > Homo sapiens < 400 > 3 gtcacatcat gacattctct ctccaaaacc cagttgttct ctgtgacgat gccataaagt 60 gacatctaat gactcaggtg tgtgtactgg gtggcagggg catgtcacac acaagcattc 120 tcaactgcag cctcagtctg ggaacgaggt ggtgcgaccc tcatttccga ggggcagctg 180 ctgttttagt tcccatctta tcattactgg aatttcaagt gtagtaccag aaaccaaatt 240 aaaattcaca ggaacgctgc actctgtact gttttcagat gttcasatat ctacaaacac 300 tcgagtagaa tcaagattag attttcaagt catcagcgtg aattaagctt tgaatcgata 360 ttcaactcag aactgagaga ggattttaca cacataatag aacatactta gccaaaaagt 420 gcacattctg tctggcactt gcgggaaaca g ccaggcaca gccagggact gcaggggtgt 480 gaggagaaag aaggcctgtc cttcagacat tgcagggctg tctaccttta aggggagtca ctctccagag 540 agattctgct ttaaccacca aaagcaataa aggatcctgt aaagggccat ttctttgtfig 600 actctgaaac cagctttaaa ccaattccaa gagatactcc gaaaatgttt 840 ttccttataa aactcacctg 660 ctgcctctgt ctgcaggtgg gctctctgaa gctgacattc atttagagaa agaaaaattc 720 caggatgttt attcaactct gtaagtctcc ttctttaaat atcttcagaa atatctttct 780 aaattcagag ttatccaaag cttacaagag gagggaagca acagaccagc ccccttatgt ggtgatggag ctgtttctag tcccagttag ctataaggca 900 aaggagagca actgccatca gcgtttgcgg taaatgctat cagaatcatg tttgaaatcc 960 tcaactctga cggttctcag aaataaaata agcaagtaac caacaacaca cacacacacc 1020 tccctccagg tctatttctc acttgtatca ttttattttt tggcaatccc aatcaattcc 1080 gct £ agccac tggatgaaaa caaaagctat gagaaatacg atgacagcat tagtactcaa 1140 agattcctca atctgtctcc ctggaacaca tttgtgtcaa aagcctgggc agtcacatga 1200 attcaagaca gtgcttcaaa aacagctgct aggataaaat actatcaata ataaagttta 1260 tgagatgagg ccagtgtttt catacttact Ctttcaacgt gca Ctgccca ttgcctttat 1320 ggagccatgg tcaaaagaaa cagctttcat caatttccag cctgaccatt tggaaactgc 1380 cagtaaacca aggctgtgaa gttggacaga agtctggaaa ctagtgttca tgagattttt 1440 tccctcattc actattatct gtcccttata ataaaaatac aagtctaaag acattaaaaa 1500 aaaaaaaaaa aa 1512

<210> 4 <211> 20 <212> DNA 15634 Sequence Listing-doc 201201819 <213> Artificial Sequence <220><223> Antisense Oligonucleotide <400> 4 gttccacctt ctcctcccag <;210>. 5 <211> 20 <212> DNA <213> Artificial sequence <220><223> Antisense oligonucleotide <400> 5 ctgctgtcct agtgtctccc <210> 6 <211> 20 <212> DNA <213> Artificial sequence <220><223> Antisense oligonucleotide <400> 6 gtctgttgct tccctcctct <210> 7 <2Π> 20 <212> DNA <;213> Artificial sequence <220><223> Antisense oligonucleotide ' <400> 7 ggagggaggt gtgtgtgtgt <210> 8 <211> 20 <212> DNA <213> Artificial sequence <220><223> Antisense Oligonucleotide <400> 8 ccttctttct cctcacaccc <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220><223> Oligonucleotide <400> 9 gccacccagt acacacacct <210> 10 <211& 20 <212> DNA <213><220> Artificial sequence-4-156341. Sequence listing.doc 201201819 <223>Antisense oligonucleotide <400> 10 gccattcgtt tcctccctga <210><211> 20 <212> DNA <213>Artificial sequence<220><223> Antisense oligonucleotide <400> 11 gctgctactc tcacccacat 156341 - Sequence Listing.doc <210> 12 < 211 > 20 <212> DNA <213> Artificial sequence <220><223> Antisense oligonucleotide <400> 12 ccagtcaggg caacacagca <210> 13 <211> 21 <212> DNA <213> A-process sequence <220><223> antisense oligonucleotide <400> 13 cctctccacg cgcagtacat t

Claims (1)

201201819 VII. Scope of Application: 1. The use of an antisense nucleotide of 5 to 30 nucleotides in length for the production of a BCL2 u (BCL2LU)-like polynucleotide in a biological system. Functional and/or performance agent, wherein the oligonucleotide has at least 50% sequence identity to a reverse complement of a natural antisense transcript of a BCL2 11 (BCL2L11)-like polynucleotide. Use of 1 wherein the natural antisense transcript comprises nucleotides 1 to 3026 of seq id NO: 2 and 5] ^ N N: 3 nucleotides to 1512. 3. Use of an antisense oligonucleotide of 5 to 30 nucleotides in length for the manufacture of a BCL2 11 (BCL2L11)-like polynucleotide for regulating a cell or tissue in a patient and/or An agent that exhibits at least 50% sequence identity to an antisense oligonucleotide of such a BCL2 11 (BCL2L11) polynucleotide. 4. The use of claim 3 wherein the natural antisense transcript comprises SEq id NO: 2 nucleotides 1 to 3 〇 26 and 8 is called id NO: 3 nucleotides 1 to 1512 〇 5 _ The use of a nucleotide that targets a region of a natural antisense oligonucleotide of a bcl2 11 (BCL2L11)-like polynucleotide, which is used for manufacturing An agent that modulates the function and/or expression of a BCL2 11 (BCL2L11)-like polynucleotide in a biological system. 6. The use of claim 5, wherein the function and/or performance of such BCL2 11 (BCL2L11) is enhanced compared to the control group. 7. The use of claim 5, wherein the antisense raised nucleotide targets the natural antisense sequence of the BCL2 11 156341.doc 201201819 (BCL2L11) polynucleotide, wherein the use of claim 5, wherein Antisense Oligonucleotide Targeting Nucleic Acid Sequences' The nucleic acid sequence includes a coding-like and/or non-coding nucleic acid sequence of BCL2 11 (BCL2L11) polynucleic acid. 9. The use of claim 5, wherein the antisense oligonucleotide targets an overlapping and/or non-overlapping sequence of a bclL2 11 (BCL2L11)-like polynucleotide. 10. The use of claim 5, wherein the antisense oligonucleotide comprises one or more modifications selected from the group consisting of at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified core Glycosylates, and combinations thereof. 11. The use of claim 1 wherein the one or more modifications comprise at least one modified sugar moiety selected from the group consisting of 2, 〇 methoxyethyl modified sugar moieties, 2'-曱An oxy-modified sugar moiety, a sugar moiety modified with 2ι_〇-alkyl, a bicyclic sugar moiety, and combinations thereof. 12. The use of claim 1 wherein the one or more modifications comprise at least one modified internucleoside linkage selected from the group consisting of phosphorothioate, 2'-methoxyethyl (ΜΟΕ) 2' - It, phosphonic acid ester, dithio acid ester, alkyl thiosulfate, amino phosphate, amino phthalate, carbonate, acid diacetate, amino acetic acid Esters, retinoyl esters, and combinations thereof. 13. The use of claim 1 , wherein the one or more modifications comprise at least one modified nucleotide selected from the group consisting of a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabinic acid (FANA), Analogs, derivatives, and combinations. 14. The use of claim 1, wherein the oligonucleotide comprises at least one oligonucleotide sequence as set forth in SEQ ID NOs: 4 to 13. 156341.doc 201201819 15. Use of a short interfering rnA (siRNA) oligonucleate of 5 to 30 nucleotides for the production of a mammalian cell or tissue such as BCL2 11 (BCL2L11) An agent for the function and/or expression of a gene, wherein the siRNA oligonucleotide is specific for an antisense polynucleotide of BCL2 11 (BCL2L11) polynucleotide, and wherein the at least one siRNA oligonucleotide The complement of at least about 5 contiguous nucleic acids in an antisense and/or sense nucleic acid molecule of such a BCL2 11 (BCL2L11) polynucleotide has at least 50% sequence identity. 16. The use of claim 15 wherein the oligonucleotide has at least about 5 contiguous nucleic acid sequences of the antisense and/or complement of the sense nucleic acid molecule of the BCL2(BCL2L11) polynucleotide. At least 8% of sequence identity. Π. The use of an antisense oligonucleotide of about 5 to 30 nucleotides in length, which is positive and/or natural against the BCL2 11 (BCL2L11) polynucleic acid. The non-coding and/or coding sequence of the sense strand is specific for the manufacture of a medicament for modulating the function and/or expression of a BCL2 11 (BCL2L11)-like cell in a mammalian cell or tissue, wherein The antisense oligonucleotide has at least 5% sequence identity with at least one of the nucleic acid sequences set forth in SEQ ID N: 1 to 3. 1 8. An oligonucleotide comprising at least one modified synthesis and modification, wherein the V, a sina is selected from the group consisting of: at least one modified sugar moiety, at least one, 'ΐ modified internucleotide linkage At least one modified nucleotide, and combinations thereof; wherein the oligonucleotide is hybridized in vivo or in vitro to a BCL2 11 (BCL2L11)-like gene and can be adjusted relative to a normal control group. 156341.doc 201201819 Class BCL2 An antisense compound and/or a sense nucleic acid molecule thereof A sequence of at least about 5 contiguous nucleic acids of a homologue, isoform, variant, derivative, mutant, fragment, or combination of complements has at least 5% sequence identity. The oligonucleotide of claim 18, wherein the nucleotide is 5 to 30 nucleotides in length and 5 to 30 contiguous nucleotides in the natural antisense transcript of the BCL2L11 gene The reverse complement sequence has a sequence identity of at least 5%. 20. The oligonucleotide of claim 19, wherein the at least one modification comprises an internucleotide linkage selected from the group consisting of: phosphorothioate, alkyl phosphonate, dithiophosphate, sulfur An alkyl phosphonate, an amino phosphate, an amine phthalic acid vinegar, a carbonated vinegar, a triacetic acid vinegar, an aminoacetic acid vinegar, a methyl vinegar, and combinations thereof. 21. The oligonucleotide of claim 19, wherein the oligonucleotide comprises at least one phosphorothioate internucleotide linkage. 22. The oligonucleotide of claim 19, wherein the oligonucleotide comprises a backbone of an internucleotide linkage of phosphorothioate. 23. The oligonucleotide of claim 19, wherein the oligonucleotide comprises at least one modified nucleotide selected from the group consisting of: a peptide nucleic acid, a locked nucleic acid (LNA), an analog thereof, Derivatives, and combinations. 24. The oligonucleotide of claim 19, wherein the oligonucleotide comprises a plurality of modifications, wherein the modifications comprise a modified nucleotide selected from the group consisting of: thio 156341.doc 201201819 acid ester, phosphonic acid Alkyl esters, dithiophosphates, alkyl thiophosphonates, amino phosphates, amino phthalates, carbonates, phosphates, amino acetates, carboxylides, and combinations thereof . 25. The oligonucleotide of claim 19, wherein the oligonucleotide comprises a plurality of modifications wherein the modifications comprise modified nucleotides selected from the group consisting of peptide nucleic acids, locked nucleic acids (LNA), analogs thereof , derivatives, and combinations. The oligonucleotide of claim 19, wherein the oligonucleotide comprises at least one modified sugar moiety selected from the group consisting of a sugar moiety modified with 2,_〇-methoxyethyl, 2'- a methoxy group-modified sugar moiety, a 2·_〇-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof. 27. The nucleotide of claim 19, wherein the oligonucleotide comprises a plurality of modifications, wherein the modifications comprise a modified oxime moiety selected from the group consisting of 2, _ 〇-methoxyethyl modified a sugar moiety, a 2,-decyloxy modified sugar moiety, a 2'-0-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof. 28. The oligonucleotide of claim 19, wherein the oligonucleotide is at least about 5 to 30 nucleotides in length and is antisense and/or sensed with a BCL2 11 (BCL2L11)-like polynucleotide. Strand hybridization wherein the oligonucleotide has at least a complement to at least about 5 contiguous nucleic acids in an antisense and/or sense encoding and/or non-coding nucleic acid sequence of such a BCL2 11 (BCL2L11) polynucleotide. About 60% of the sequence '--. 29. The oligonucleotide of claim 19, wherein the oligonucleotide has at least about 5 of the antisense and/or sense encoding and/or non-coding nucleic acid sequences of the BCL2 11 (BCL2L11) polynucleotide. The complementary sequence of contiguous nucleic acids has a sequence identity of at least 156341.doc 201201819 of about 80%. 30. The oligonucleotide of claim 19, wherein the oligonucleotide hybridizes with at least one BCL2 11 (BCL2L11) polynucleic acid in vivo or in vitro, and the class can be adjusted relative to a normal control group. The performance and/or function of the BCL2 η (BCL2L11) polynucleotide. 31. The oligonucleotide of claim 19, wherein the oligonucleotide comprises the sequence set forth in SEQ m NO: 4 to 13. 32. A pharmaceutical composition comprising one or more oligonucleotides specific for one or more of the BCL2 11 (BCL2L11) polynucleotides of claim 18 and a pharmaceutically acceptable excipient. 33. The composition of claim 32, wherein the oligonucleotides are aligned with any one of the nucleotide sequences set forth as SEq ID NO: 4 to 13, having a sequence identity of at least about 40%. 34. The composition of claim 32, wherein the oligonucleotides comprise a nucleotide sequence as set forth in SEq ID NO: 4 to 13. 35. The composition of claim 34 wherein the oligonucleotides such as SEq m NO: 4 to 13 comprise one or more modifications or substitutions. 36. The composition of claim 35, wherein the one or more modifications are selected from the group consisting of: phosphorothioates, phosphonium phosphonates, peptide nucleic acids, locked nucleic acid (LNA) molecules, and combinations thereof. 37. Use of an antisense oligonucleotide for the manufacture of a medicament for preventing or treating a disease associated with at least one BCL2 11 (BCL2L11)-like polynucleotide and/or at least one of its encoded products, wherein The antisense oligonucleotide and the natural anti-I56341.doc 201201819 sequence of the at least one BCL2 11 (BCL2L11)-like polynucleotide, ασ, and modulate the at least one BCL2 11 (BCL2L11)-like polynucleotide which performed. 38. The use of claim 37, wherein the disease associated with the at least one BCL2! J (BCL2L11) polynucleic acid is selected from the group consisting of: a disease associated with BeL2Ln and/or a performance-related disease or A condition, cancer, abnormal cell apoptosis proliferative disease or condition, a disease or condition associated with a mitochondrial cell apoptosis pathway, leukemia, an autoimmune disease or condition, a disease or condition associated with an immune response, an infection Hyperproliferative disorders, hyperproliferation of reproductive tissues (eg uterine cancer, testicular and ovarian cancer, endometriosis, squamous cervix, glandular epithelial cancer, etc.), neurological diseases or conditions, and Developmental disorders related to developmental embryos, blood diseases or conditions, impaired tissue homeostasis, rheumatoid arthritis, septicemia, retinal ganglion cell death, liver disease, and kidney disease. 39. Use of a nucleotide of 5 to 30 nucleotides in length for the manufacture of an agent for inducing apoptosis in a biological system, wherein the oligonucleotide is polymerized with BCL2L11 in the system At least 5% of the 5 to 30 contiguous nucleotides within the reverse complement of the natural antisense transcript of the nucleotide are at least 5% identical. 40. The use of claim 39, wherein the natural antisense transcript has a 卩 id NO: 2 or 3. 41 ‘The use of claim 39, wherein the biological system is a patient cell or tissue. 42. Use of an oligonucleotide of about 5 to 30 nucleotides in length for the manufacture of an agent for inducing apoptosis in a biological system, wherein the oligonucleotide can be targeted by The natural antisense transcript of the target gene upregulates the 156341.doc 201201819 gene, and wherein the oligonucleotide is at least 5% identical to the reverse complement of the natural antisense transcript of the upregulated gene. 43. A method for identifying and selecting at least one oligonucleotide selective for a natural antisense transcript of a BCL2L11 gene as a selected target polynucleotide for use in an in vivo technique, the method comprising: identifying at least one An oligonucleotide of at least 5 contiguous nucleotides at least partially complementary to an antisense polynucleotide of the selected target polynucleotide; measuring the antisense oligonucleotide and the target polynucleoside under stringent hybridization conditions The thermal melting point of the acid or the hybrid of the antisense polynucleotide of the selected target polynucleotide; and selecting at least one oligonucleotide for in vivo administration based on the information obtained. 44. A method of modulating the function and/or expression of a bcl2 11 (BC:L2L11)-like polynucleotide in a biological system in vitro comprising: bringing the system to a length of 5 to 30 nucleotides The antisense oligonucleotide is contacted, wherein the at least one oligonucleotide has at least 50% sequence identity with the reverse complement of the natural antisense transcript of the bcL2 11 (BCL2L11) polynucleoside; This modulates the function and/or expression of such a BCL2 u (BCL2L11) polynucleotide. 45. The function and/or expression of a bcL2 11 (BCL2L11)-like polynucleotide in an in vitro regulatory biological system according to claim 44 A method comprising: contacting the biological system with at least one antisense oligonucleotide of 5 to 30 nucleotides in length, wherein the at least one oligonucleotide comprises a natural antisense transcript comprising the SEQ ID NO: The reverse complement of the nucleotides of nucleotides 1 to 3026 of 2 and 5 to 30 contiguous nucleotides of nucleotides 1 to 15 12 of SEQ ID NO: 3 has at least 50% sequence identity The function and/or performance of the 156341.doc 201201819 BCL2 11 (BCL2L11) polynucleotide is thereby regulated. 46. A method of modulating the function and/or expression of a BCL2 11 (BCL2L11)-like polynucleotide in a patient cell or tissue in vitro, comprising: Having the cells or tissues at least 5 to 30 in length An antisense oligonucleotide contact of a nucleotide, wherein the oligonucleotide has at least 50% sequence identity to an antisense oligonucleotide of the BCL2 11 (BCL2L11) polynucleotide; thereby in vivo The function and/or expression of such a BCL2 11 (BCL2L11) polynucleotide in a patient's cells or tissues is modulated externally. 47. The method of claim 46, wherein the function and/or performance of a BCL2 11 (BCL2L11)-like polynucleotide in a patient cell or tissue is modulated, comprising: causing the sigma with at least one length of 5 to 30 Contact with an antisense oligonucleotide of nucleotide acid, wherein the at least one oligonucleotide and the nucleotides comprising nucleotides 1 to 3026 and SEQ ID NO: 3 of the natural antisense transcript of SEQ ID NO: 2 The reverse complement of 5 to 30 consecutive nucleotides of nucleotides from 1 to 1512 has a sequence identity of at least 50%; thereby modulating the function of the BCL2 11 (BCL2L11) polynucleic acid and/or Or performance. 48. A method of modulating the function and/or expression of a bclL2 11 (BCL2L11)-like polynucleotide in a biological system in vitro comprising: targeting the system to at least one of the BCL2 11 (BCL2L11) polynuclear The antisense oligonucleotides of the region of the natural antisense oligonucleotide of the nucleoside are contacted; thereby modulating the function and/or performance of such a BCL2 11 (BCL2L11) polynucleotide. 49. The method of claim 48, wherein the function and/or performance of the BCL2 11 (BCL2L11) in vitro is enhanced over the control group. 50. The method of claim 48, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a 156341.doc 201201819 BCL2 11 (BCL2L11) polynucleotide. The method of claim 48, wherein the at least one antisense oligonucleotide targets a nucleic acid sequence comprising a BCL2 11 (Bcl2Ui)-like polynucleotide encoding and/or non-coding nucleic acid sequence. 52. The method of claim 48, wherein the at least one antisense oligonucleotide targets an overlapping and/or non-overlapping sequence of a BCL2(R)-like (BCL2L11) polynucleic acid. 53. The method of claim 48, wherein the at least one antisense oligonucleotide comprises one or more modifications selected from the group consisting of at least one modified sugar moiety, at least one modified internucleoside linkage, at least one Modified nucleotides, and combinations thereof. 54. The method of claim 53, wherein the one or more modifications comprise at least one modified sugar moiety selected from the group consisting of 2,-〇-methoxyethyl modified sugar moiety, 2,-methoxy a sugar moiety modified by a base, a sugar moiety modified with 2, 〇·alkyl, a bicyclic sugar moiety, and combinations thereof. 55. The method of claim 53, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from the group consisting of: phosphorothioate, 2,-methoxyethyl (ΜΟΕ), 2 , fluorine, alkyl phosphonate, dithiophosphate, thiophosphonate, amino aryl ester, urethane, carbonate, acid-depleted triacetate, amino acetate, Carboxynonyl esters, and combinations thereof. 56. The method of claim 53, wherein the one or more modifications comprise at least one modified nucleotide selected from the group consisting of a peptide nucleic acid (ρΝΑ), a locked nucleic acid (LNA), an arabinic acid (FANA), and the like The method of claim 44, wherein the at least one nucleus acid comprises at least one oligonucleotide sequence as set forth in SEQ ID NOS: 4 to 13. 58. A method of modulating the function and/or expression of a bclL2! j (BCL2L11) gene in a mammalian cell or tissue in vitro, comprising: equating the cell or tissue with at least one of 5 to 3 lengths Nucleotide short interfering RNA (siRNA) nucleotide contacts, the at least one siRNA raised nucleotide specific for an antisense polynucleotide of a BCL2 11 (BCL2L11)-like polynucleotide, wherein the at least one The siRNA oligonucleotide has at least 5% sequence identity to the antisense of the BCL2 11 (BCL2L11) polynucleotide and/or the complement of at least about 5 contiguous nucleic acids in the sense nucleic acid molecule, and in vivo The function and/or expression of BCL2 11 (BCL2L11) in mammalian cells or tissues is regulated externally. 59. The method of claim 58, wherein the nucleotide and the sequence of at least about 5 contiguous nucleic acids of the antisense and/or complement of the sense nucleic acid molecule of the bcl2 11 (BCL2L11) polynucleotide have At least 8% of sequence identity. 60. A method of modulating the function and/or expression of a bcl2 11 (BCL2L11)-like cell in a mammalian cell or tissue in vitro comprising: bringing the cells or tissues to at least one of about 5 to 30 cores in length Antisense oligodeoxynucleotide contact with glucosinolates. The antisense oligonucleotide is specific for non-coding and/or coding sequences of sense-like and/or natural antisense strands of BCL2 11 (BCL2L11)-like polynucleotides. Wherein the at least one antisense oligonucleotide has at least 5% sequence identity with at least one of the nucleic acid sequences set forth in SEQ ID NOS: 1 to 3; and modulating the mammalian cell or tissue in vitro Function and/or performance of BCL2 11 (BCL2L11). 156341.doc