siRNA compound that inhibits expression of APOC3

Nucleic acid products and compositions and their uses are provided. In particular, nucleic acid products are provided that modulate, interfere with, or inhibit APOC3 gene expression. The products can be oligomeric compounds that comprise at least a first region of linked nucleosides having at least a first nucleobase sequence that is at least partially complementary to at least a portion of RNA transcribed from a APOC3 gene, wherein said first nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs 1 to 39

1. 11. An siRNA compound that inhibits expression of APOC3, comprising a single strand of 33 or 34 nucleobases that is modified or unmodified, wherein said single strand comprises: (i) a first nucleobase sequence directly linked to (ii) a second nucleobase sequence, wherein said first and second nucleobase sequences form a hairpin loop consisting of a duplex region of 14 or 15 base pairs, and a loop consisting of 4 or 5 nucleosides, wherein said first and second nucleobase sequences in unmodified form are, respectively, SEQ ID NOs: 23 and 423; 24 and 424; 28 and 428; 29 and 429; 31 and 431; 89 and 489; 90 and 490; 94 and 494; 117 and 517; 121 and 521; 128 and 528; 137 and 537; 138 and 538; 148 and 548; 149 and 549; 167 and 567; 171 and 571; 175 and 575; 185 and 585; 191 and 591; 193 and 593; 206 and 606; 209 and 609; 212 and 612; 213 and 613; 217 and 617; 218 and 618; 219 and 619; 220 and 620; 221 and 621; 223 and 623; 225 and 625; 254 and 654; 262 and 662; 271 and 671; 272 and 672; 274 and 674; 275 and 675; 276 and 676; 277 and 677; 278 and 678; 280 and 680; 281 and 681; 282 and 682; 283 and 683; 285 and 685; 286 and 686; 291 and 691; 293 and 693; 296 and 696; 297 and 697; 299 and 699; 300 and 700; 303 and 703; 324 and 724; 328 and 728; 331 and 731; 332 and 732; 334 and 734; 336 and 736; 337 and 737; 338 and 738; 339 and 739; 340 and 740; 341 and 741; 342 and 742; 343 and 743; 345 and 745; 346 and 746; 347 and 747; 366 and 766; 367 and 767; 369 and 769; 370 and 770; 368 and 768; 372 and 772; 373 and 773.
2. 22. The compound according to claim 1, wherein said first nucleobase sequence and said second sequence are selected from: SEQ ID NOs: 28 and 428; 137 and 537; 149 and 549; 167 and 567; 175 and 575; 185 and 585; 191 and 591; 193 and 593; 221 and 621; 225 and 625; 254 and 654; 262 and 662; 271 and 671; 274 and 674; 277 and 677; 280 and 680; 286 and 686; 293 and 693; 297 and 697; 328 and 728; 332 and 732; 334 and 734; 336 and 736; 337 and 737; 343 and 743; 366 and 766; 367 and 767; 369 and 769; and 373 and 773.
3. 33. The compound according to claim 2, wherein said first nucleobase sequence and said second nucleobase sequence are selected from the group consisting of: SEQ ID NOs: 28 and 428, 277 and 677, 336 and 736, 337 and 737, 366 and 766, 367 and 767, and 369 and 769.
4. 44. The compound according to claim 1, further comprising one or more ligands, wherein said one or more ligands comprise one or more N-Acetyl-Galactosamine moieties.

CROSS REFERENCE TO RELATED PATENT APPLICATIONS
This application claims the benefit of and priority to two U.S. Provisional Patent Applications, Nos. 63/214,608, filed Jun. 24, 2021, and 63/318,287, filed Mar. 9, 2022, the contents of which are incorporated herein by reference in their entirety.


SEQUENCE LISTING
The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Nov. 7, 2022, is named 4690_0050C_SL_ST.25.txt and is 373 kilobytes in size.
FIELD
Nucleic acid products and compositions, and their uses, that modulate, in particular interfere with, or inhibit, apolipoprotein C3 (APOC3) gene expression are provided. Specific embodiments provide methods, compounds, and compositions for reducing expression of APOC3 mRNA and protein in an animal. Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate APOC3-associated disorders such as dyslipidemia, hypertriglyceridemia, hyperchylomicronemia, and atherosclerotic cardiovascular disease (ASCVD).
BACKGROUND
Triglycerides are esters of glycerol with three fatty acids. They serve as storage of fat and energy and are transported via the bloodstream. Excess level of blood triglycerides have been recognized early on as causative agents or bystanders of a range of disorders. More recent evidence suggests a causative role, partly in conjunction with elevated levels of cholesterol (in particular LDL cholesterol) in ASCVD and disorders subsumed under this term or associated therewith. A more comprehensive list of disorders associated with elevated levels of triglycerides is given in the embodiments disclosed further below. Apolipoprotein C3 is secreted by the liver and the small intestine. It can be found on triglyceride-rich lipoproteins including very low density lipoproteins (VLDL) and chylomicrons. It is involved in the negative regulation of lipid catabolism, especially triglyceride catabolism, and of the clearance of VLDL, LDL and HDL lipoproteins. A molecular function of APOC3 is the inhibition of lipoprotein lipase and of hepatic lipase.
Disease
Abnormal amounts of circulating triglycerides, also referred to as hypertriglyceridemia, is a recognized disorder in itself which is inter alia owed to the fact that such abnormal amounts, in particular if they persist over extended periods of time, may entail disorders of the cardiovascular system and/or inflammation.
Treatment
Established treatments include the administration of statins such as Rosuvastatin and Simvastatin as well as of fibrates such as fenofibrate. However, statins may cause side effects, and certain patients are statin-intolerant.
There therefore remains a need for therapies to treat APOC3-associated diseases. We, therefore, aim to provide compounds, methods, and pharmaceutical compositions for the treatment of such diseases. Double-stranded RNA (dsRNA) able to complementarily bind expressed mRNA has been shown to be able to block gene expression (Fire et al., 1998, Nature. 1998 Feb. 19; 391 (6669):806-1 1 and Elbashir et at., 2001, Nature. 2001 May 24; 41 1 (6836):494-8) by a mechanism that has been termed RNA interference (RNAi). Short dsRNAs direct gene-specific, post-transcriptional silencing in many organisms, including vertebrates, and have become a useful tool for studying gene function. RNAi is mediated by the RNA-induced silencing complex (RISC), a sequence-specific, multi-component nuclease that destroys messenger RNAs homologous to the silencing trigger loaded into the RISC complex. Interfering RNA (iRNA) such as siRNAs, antisense RNA, and micro-RNA are oligonucleotides that prevent the formation of proteins by gene-silencing i.e. inhibiting gene translation of the protein through degradation of mRNA molecules. Gene-silencing agents are becoming increasingly important for therapeutic applications in medicine.
According to Watts and Corey in the Journal of Pathology (2012; Vol 226, p 365-379) there are algorithms that can be used to design nucleic acid silencing triggers, but all of these have severe limitations. It may take various experimental methods to identify potent siRNAs, as algorithms do not take into account factors such as tertiary structure of the target mRNA or the involvement of RNA binding proteins. Therefore the discovery of a potent nucleic acid silencing trigger with minimal off-target effects is a complex process. For the pharmaceutical development of these highly charged molecules it is necessary that they can be synthesised economically, distributed to target tissues, enter cells and function within acceptable limits of toxicity. An aim is to, therefore, provide compounds, methods, and pharmaceutical compositions for the treatment of thromboembolic diseases as described herein, which comprise oligomeric compounds that modulate, in particular inhibit, gene expression by RNAi.
SUMMARY
Nucleic acid products are provided that modulate, in particular, interfere with or inhibit, apolipoprotein C3 (APOC3) gene expression, and associated therapeutic uses. Specific oligomeric compounds and sequences are described herein. This summary provides a simplified form that is further described below in the detailed description. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to determine the scope of the claimed subject matter.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1a shows dose curves of APOC3 leads for candidates in primary human hepatocytes;
FIG. 1b shows dose curves of APOC3 leads for Humanized mouse study in primary human hepatocytes;
FIG. 2 shows a timeline including the time point of applying the dose to the mice and time points for taking samples;
FIG. 3 shows remaining liver APOC3 mRNA and plasma APOC3 protein levels for the animals treated with APOC3-targeting mxRNA constructs as compared to the control animals;
FIG. 4 shows serum triglycerides and total cholesterol in the serum of the animals treated with APOC3-targeting mxRNA constructs as compared to the control (PBS);
FIG. 5a shows a Mean Percent of remaining APOC3 mRNA in liver tissues in plasma measured using ELISA for the animals treated with APOC3-targeting mxRNA constructs (10 mg/kg) as compared to the control animals;
FIG. 5b shows APOC3 protein levels in plasma measured using ELISA for the animals treated with APOC3-targeting mxRNA constructs (10 mg/kg) as compared to the control animals;
FIG. 6a shows the mean percent of triglycerides (TG) in the serum of the animals treated with APOC3 targeting mxRNA constructs as compared to the control animals at weeks 2 and 6;
FIG. 6b shows the total cholesterol (TC) level in serum of animals treated with APOC3 targeting mxRNA constructs as compared to the control animals at weeks 2 and 6;
FIG. 7 prevents a schematic overview of the duration study performed with compound A28(14-4)mF (also designated STP125G) in mice with a humanized liver;
FIG. 8a shows APOC3 mRNA as a function of time as observed in the duration study between control and treatment groups;
FIG. 8b shows APOC3 protein knockdown as a function of time as observed in the duration study between control and treatment groups;
FIG. 9a show serum triglyceride levels as a function of time between control and treatment groups;
FIG. 9b show serum total cholesterol levels as a function of time between control and treatment groups; and
FIG. 10 illustrates the humanized liver of the mice used for the duration study.



DETAILED DESCRIPTION AND EMBODIMENTS
The following are non-limiting aspects:
Aspect 1. An oligomeric compound capable of inhibiting expression of APOC3, wherein said compound comprises at least a first region of linked nucleosides having at least a first nucleobase sequence that is at least partially complementary to at least a portion of RNA transcribed from a APOC3 gene, wherein said first nucleobase sequence is selected from the following sequences, or a portion thereof: sequences of SEQ ID NOs 1 to 391, wherein said portion preferably has a length of at least 18 nucleotides.
Particularly preferred embodiments relate to mxRNAs: for further details see the embodiments and their discussion further below.
In addition, the antisense and sense regions disclosed herein may serve as building blocks for compounds which are directed to multiple targets. The general architecture of such compound ds is described in WO2020/065602.
Furthermore, and as disclosed further below, the disclosed embodiments also relate to double-stranded RNAs (dsRNAs). In contrast to an mxRNA, which has a hairpin-like structure connecting the sense and antisense RNA strands, a dsRNA lacks the hairpin loop and, therefore, dsRNA comprises two strands.
Aspect 2. A composition comprising an oligomeric compound according to aspect 1, and a physiologically acceptable excipient.
Aspect 3. A pharmaceutical composition comprising an oligomeric compound according to aspect 1.
Aspect 4. An oligomeric compound according to aspect 1, for use in human or veterinary medicine or therapy.
Aspect 5. An oligomeric compound according to aspect 1, for use in a method of treating a disease or disorder.
Aspect 6. A method of treating a disease or disorder comprising administration of an oligomeric compound according to aspect 1, to an individual in need of treatment.
Aspect 7. Use of an oligomeric compound according to aspect 1, for use in research as a gene function analysis tool.
Aspect 8. Use of an oligomeric compound according to aspect 1 in the manufacture of a medicament for a treatment of a disease or disorder.
Further embodiments are described below by way of example only. These examples represent the best ways of putting the disclosed embodiments into practice that are currently known to the applicant, although they are not the only ways in which this could be achieved.
It will be understood that the benefits and advantages described herein may relate to one embodiment or may relate to several embodiments. The embodiments are not limited to those that solve any or all of the stated problems or those that have any or all of the stated benefits and advantages.
Features of different aspects and embodiments as described herein may be combined as appropriate, as would be apparent to a skilled person, and may be combined with any other aspects.
Definitions
The following definitions pertain to the disclosed embodiments throughout. In many instances, the definitions, in addition to the respective definition as such, provide non-exhaustive listings of possible implementations, which amount to preferred embodiments.
Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, and chemical analysis. Certain such techniques and procedures may be found for example in “Carbohydrate Modifications in Antisense Research” Edited by Sangvi and Cook, American Chemical Society, Washington D.C., 1994; “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., 21st edition, 2005; and “Antisense Drug Technology, Principles, Strategies, and Applications” Edited by Stanley T. Crooke, CRC Press, Boca Raton, Fla.; and Sambrook et al., “Molecular Cloning, A laboratory Manual,” 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, which are hereby incorporated by reference for any purpose. Where permitted, all patents, applications, published applications and other publications and other data referred to throughout in the disclosure are incorporated by reference herein in their entirety.
Unless otherwise indicated, the following terms have the following meanings:
As used herein, “excipient” means any compound or mixture of compounds that is added to a composition as provided herein that is suitable for delivery of an oligomeric compound.
As used herein, “nucleoside” means a compound comprising a nucleobase moiety and a sugar moiety. Nucleosides include, but are not limited to, naturally occurring nucleosides (as found in DNA and RNA) and modified nucleosides. Nucleosides may be linked to a phosphate moiety, phosphate-linked nucleosides also being referred to as “nucleotides”.
As used herein, “chemical modification” or “chemically modified” means a chemical difference in a compound when compared to a naturally occurring counterpart. Chemical modifications of oligonucleotides include nucleoside modifications (including sugar moiety modifications and nucleobase modifications) and internucleoside linkage modifications. In reference to an oligonucleotide, chemical modification does not include differences only in nucleobase sequence.
As used herein, “furanosyl” means a structure comprising a 5-membered ring comprising four carbon atoms and one oxygen atom.
As used herein, “naturally occurring sugar moiety” means a ribofuranosyl as found in naturally occurring RNA or a deoxyribofuranosyl as found in naturally occurring DNA. A “naturally occurring sugar moiety” as referred to herein is also termed as an “unmodified sugar moiety”. In particular, such a “naturally occurring sugar moiety” or an “unmodified sugar moiety” as referred to herein has a —H (DNA sugar moiety) or —OH(RNA sugar moiety) at the 2′-position of the sugar moiety, especially a —H (DNA sugar moiety) at the 2′-position of the sugar moiety.
As used herein, “sugar moiety” means a naturally occurring sugar moiety or a modified sugar moiety of a nucleoside. As used herein, “modified sugar moiety” means a substituted sugar moiety or a sugar surrogate.
As used herein, “substituted sugar moiety” means a furanosyl that has been substituted. Substituted sugar moieties include, but are not limited to furanosyls comprising substituents at the 2′-position, the 3′-position, the 5′-position and/or the 4′-position. Certain substituted sugar moieties are bicyclic sugar moieties.
As used herein, “2′-substituted sugar moiety” means a furanosyl comprising a substituent at the 2′-position other than H or OH. Unless otherwise indicated, a 2′-substituted sugar moiety is not a bicyclic sugar moiety (i.e., the 2′-substituent of a 2′-substituted sugar moiety does not form a bridge to another atom of the furanosyl ring).
As used herein, “MOE” means —OCH2CH2OCH3.
As used herein, “2′-F nucleoside” refers to a nucleoside comprising a sugar comprising fluorine at the 2′ position. Unless otherwise indicated, the fluorine in a 2′-F nucleoside is in the ribo position (replacing the OH of a natural ribose). Duplexes of uniformly modified 2′-fluorinated (ribo) oligonucleotides hybridized to RNA strands are not RNase H substrates while the ara analogs retain RNase H activity.
As used herein the term “sugar surrogate” means a structure that does not comprise a furanosyl and that is capable of replacing the naturally occurring sugar moiety of a nucleoside, such that the resulting nucleoside sub-units are capable of linking together and/or linking to other nucleosides to form an oligomeric compound which is capable of hybridizing to a complementary oligomeric compound. Such structures include rings comprising a different number of atoms than furanosyl (e.g., 4, 6, or 7-membered rings); replacement of the oxygen of a furanosyl with a non-oxygen atom (e.g., carbon, sulfur, or nitrogen); or both a change in the number of atoms and a replacement of the oxygen. Such structures may also comprise substitutions corresponding to those described for substituted sugar moieties (e.g., 6-membered carbocyclic bicyclic sugar surrogates optionally comprising additional substituents). Sugar surrogates also include more complex sugar replacements (e.g., the non-ring systems of peptide nucleic acid). Sugar surrogates include without limitation morpholinos, cyclohexenyls and cyclohexitols.
As used herein, “bicyclic sugar moiety” means a modified sugar moiety comprising a 4 to 7 membered ring (including but not limited to a furanosyl) comprising a bridge connecting two atoms of the 4 to 7 membered ring to form a second ring, resulting in a bicyclic structure. In certain embodiments, the 4 to 7 membered ring is a sugar ring. In certain embodiments the 4 to 7 membered ring is a furanosyl. In certain such embodiments, the bridge connects the 2 ‘-carbon and the 4’-carbon of the furanosyl.
As used herein, “nucleotide” means a nucleoside further comprising a phosphate linking group. As used herein, “linked nucleosides” may or may not be linked by phosphate linkages and thus includes, but is not limited to “linked nucleotides.” As used herein, “linked nucleosides” are nucleosides that are connected in a continuous sequence (i.e. no additional nucleosides are present between those that are linked).
As used herein, “nucleobase” means a group of atoms that can be linked to a sugar moiety to create a nucleoside that is capable of incorporation into an oligonucleotide, and wherein the group of atoms is capable of bonding, more specifically hydrogen bonding, with a complementary naturally occurring nucleobase of another oligonucleotide or nucleic acid. Nucleobases may be naturally occurring or may be modified.
As used herein the terms, “unmodified nucleobase” or “naturally occurring nucleobase” means the naturally occurring heterocyclic nucleobases of RNA or DNA: the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) (including 5-methyl C), and uracil (U).
As used herein, “modified nucleobase” means any nucleobase that is not a naturally occurring nucleobase.
As used herein, “modified nucleoside” means a nucleoside comprising at least one chemical modification compared to naturally occurring RNA or DNA nucleosides. Modified nucleosides can comprise a modified sugar moiety and/or a modified nucleobase.
As used herein, “bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety. As used herein, “locked nucleic acid nucleoside” or “LNA” means a nucleoside comprising a bicyclic sugar moiety comprising a 4′—CH2—O-2′bridge.
As used herein, “2 ‘-substituted nucleoside” means a nucleoside comprising a substituent at the 2’-position of the sugar moiety other than H or OH. Unless otherwise indicated, a 2 ′-substituted nucleoside is not a bicyclic nucleoside.
As used herein, “deoxynucleoside” means a nucleoside comprising 2′—H furanosyl sugar moiety, as found in naturally occurring deoxyribonucleosides (DNA). In certain embodiments, a 2′-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (e.g., uracil).
As used herein, “oligonucleotide” means a compound comprising a plurality of linked nucleosides. In certain embodiments, an oligonucleotide comprises one or more unmodified ribonucleosides (RNA) and/or unmodified deoxyribonucleosides (DNA) and/or one or more modified nucleosides.
As used herein, “modified oligonucleotide” means an oligonucleotide comprising at least one modified nucleoside and/or at least one modified internucleoside linkage.
Preferred modified internucleoside linkages are those which confer increased stability as compared to the naturally occurring phosphodiesters. “Stability” means, in particular, the stability against hydrolysis including enzyme-catalyzed hydrolysis, enzymes including exonucleases and endonucleases.
Preferred positions for such modified internucleoside linkages include the termini and the hairpin loop of single-stranded oligomeric compounds. For example, the internucleoside linkages connecting first and second nucleoside and second and third nucleoside counting from the 5′ terminus, and/or the internucleoside linkages connecting first and second nucleoside and second and third nucleoside counting from the 3′ terminus are modified. In addition, a linkage connecting the terminal nucleoside of the 3′ terminus with a ligand, such as GaINAc, may be modified.
As discussed above, preferred positions are in the hairpin loop of said single-stranded oligomeric compounds. In particular, all linkages, all but one linkages or the majority of linkages in the hairpin loop are modified. As used herein, “linkages in the hairpin loop” designates the linkages between nucleosides which are not engaged in base pairing. For example, in a hairpin loop consisting of five nucleosides, there are four linkages between nucleosides which are not engaged in base pairing. Preferably, the term “linkages in the hairpin loop” also extends to the linkages connecting the stem to the loop, i.e., those linkages which connect a base-paired nucleoside to a non-based paired nucleoside. Generally, there are two such positions in hairpins and mxRNAs as described herein.
Most preferred is that modified internucleoside linkages are at both termini and in the hairpin loop. As used herein, “linkage” or “linking group” means a group of atoms that link together two or more other groups of atoms.
As used herein “internucleoside linkage” means a covalent linkage between adjacent nucleosides in an oligonucleotide.
As used herein “naturally occurring internucleoside linkage” means a 3′ to 5′ phosphodiester linkage. As used herein, “modified internucleoside linkage” means any internucleoside linkage other than a naturally occurring internucleoside linkage. In particular, a “modified internucleoside linkage” as referred to herein can include a modified phosphorous linking group such as a phosphorothioate or phosphorodithioate internucleoside linkage.
As used herein, “terminal internucleoside linkage” means the linkage between the last two nucleosides of an oligonucleotide or defined region thereof.
As used herein, “phosphorus linking group” means a linking group comprising a phosphorus atom and can include naturally occurring phosphorous linking groups as present in naturally occurring RNA or DNA, such as phosphodiester linking groups, or modified phosphorous linking groups that are not generally present in naturally occurring RNA or DNA, such as phosphorothioate or phosphorodithioate linking groups. Phosphorus linking groups can therefore include without limitation, phosphodiester, phosphorothioate, phosphorodithioate, phosphonate, methylphosphonate, phosphoramidate, phosphorothioamidate, thionoalkylphosphonate, phosphotriesters, thionoalkylphosphotriester and boranophosphate.
As used herein, “internucleoside phosphorus linking group” means a phosphorus linking group that directly links two nucleosides.
As used herein, “oligomeric compound” means a polymeric structure comprising two or more substructures. In certain embodiments, an oligomeric compound comprises an oligonucleotide, such as a modified oligonucletide. In certain embodiments, an oligomeric compound further comprises one or more conjugate groups and/or terminal groups and/or ligands. In certain embodiments, an oligomeric compound consists of an oligonucleotide. In certain embodiments, an oligomeric compound comprises a backbone of one or more linked monomeric sugar moieties, where each linked monomeric sugar moiety is directly or indirectly attached to a heterocyclic base moiety. In certain embodiments, oligomeric compounds may also include monomeric sugar moieties that are not linked to a heterocyclic base moiety, thereby providing abasic sites. Oligomeric compounds may be defined in terms of a nucleobase sequence only, i.e., by specifying the sequence of A, G, C, U (or T). In such a case, the structure of the sugar-phosphate backbone is not particularly limited and may or may not comprise modified sugars and/or modified phosphates. On the other hand, oligomeric compounds may be more comprehensively defined, i.e, by specifying not only the nucleobase sequence, but also the structure of the backbone, in particular the modification status of the sugars (unmodified, 2′-0Me modified, 2′-F modified etc.) and/or of the phosphates.
As used herein, “terminal group” means one or more atom attached to either, or both, the 3′ end or the 5′ end of an oligonucleotide. In certain embodiments, a terminal group comprises one or more terminal group nucleosides.
As used herein, “conjugate” or “conjugate group” means an atom or group of atoms bound to an oligonucleotide or oligomeric compound. In certain embodiments, a conjugate group links a ligand to a modified oligonucleotide or oligomeric compound. In general, conjugate groups can modify one or more properties of the compound to which they are attached, including, but not limited to pharmacodynamic, pharmacokinetic, binding, absorption, cellular distribution, cellular uptake, charge and/or clearance properties.
As used herein, “conjugate linker” or “linker” in the context of a conjugate group means a portion of a conjugate group comprising any atom or group of atoms and which covalently link an oligonucleotide to another portion of the conjugate group. In certain embodiments, the point of attachment on the oligomeric compound is the 3 ′-oxygen atom of the 3′-hydroxyl group of the 3′ terminal nucleoside of the oligonucleotide. In certain embodiments the point of attachment on the oligomeric compound is the 5′-oxygen atom of the 5′-hydroxyl group of the 5′ terminal nucleoside of the oligonucleotide. In certain embodiments, the bond for forming attachment to the oligomeric compound is a cleavable bond. In certain such embodiments, such cleavable bond constitutes all or part of a cleavable moiety.
In certain embodiments, conjugate groups comprise a cleavable moiety (e.g., a cleavable bond or cleavable nucleoside) and ligand portion that can comprise one or more ligands, such as a carbohydrate cluster portion, such as an N-Acetyl-Galactosamine, also referred to as “GaINAc”, cluster portion. In certain embodiments, the carbohydrate cluster portion is identified by the number and identity of the ligand. For example, in certain embodiments, the carbohydrate cluster portion comprises 2 GaINAc groups. For example, in certain embodiments, the carbohydrate cluster portion comprises 3 GaINAc groups and this is particularly preferred. In certain embodiments, the carbohydrate cluster portion comprises 4 GaINAc groups. Such ligand portions are attached to an oligomeric compound via a cleavable moiety, such as a cleavable bond or cleavable nucleoside. The ligands can be arranged in a linear or branched configuration, such as a biantennary or triantennary configurations. A preferred carbohydrate cluster, also referred to as “toothbrush,” has the following formula:




wherein in said structural formula one, two, or three phosphodiester linkages can also be substituted by phosphothionate linkages.

As used herein, “cleavable moiety” means a bond or group that is capable of being cleaved under physiological conditions. In certain embodiments, a cleavable moiety is cleaved inside a cell or sub-cellular compartments, such as an endosome or lysosome. In certain embodiments, a cleavable moiety is cleaved by endogenous enzymes, such as nucleases. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is a phosphodiester linkage.
As used herein, “cleavable bond” means any chemical bond capable of being broken.
As used herein, “carbohydrate cluster” means a compound having one or more carbohydrate residues attached to a linker group.
As used herein, “modified carbohydrate” means any carbohydrate having one or more chemical modifications relative to naturally occurring carbohydrates.
As used herein, “carbohydrate derivative” means any compound which may be synthesized using a carbohydrate as a starting material or intermediate.
As used herein, “carbohydrate” means a naturally occurring carbohydrate, a modified carbohydrate, or a carbohydrate derivative. A carbohydrate is a biomolecule including carbon (C), hydrogen (H) and oxygen (O) atoms. Carbohydrates can include monosaccharide, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides or polysaccharides, such as one or more galactose moieties, one or more lactose moieties, one or more N-Acetyl-Galactosamine moieties, and/or one or more mannose moieties. A particularly preferred carbohydrate is N-Acetyl-Galactosamine.
As used herein, “strand” means an oligomeric compound comprising linked nucleosides.
As used herein, “single strand” or “single-stranded” means an oligomeric compound comprising linked nucleosides that are connected in a continuous sequence without a break therebetween. Such single strands may include regions of sufficient self-complementarity so as to be capable of forming a stable self-duplex in a hairpin structure.
As used herein, “hairpin” means a single stranded oligomeric compound that includes a duplex formed by base pairing between sequences in the strand that are self-complementary and opposite in directionality.
As used herein, “hairpin loop” means an unpaired loop of linked nucleosides in a hairpin that is created as a result of hybridization of the self-complementary sequences. The resulting structure looks like a loop or a U-shape.
In particular, short hairpin RNA, also denoted as shRNA, comprises a duplex region and a loop connecting the regions forming the duplex. The end of the duplex region which does not carry the loop may be blunt-ended or carry (a) 3′ and/or (a) 5′ overhang(s). Preference is given to blunt-ended constructs.
As used herein, “directionality” means the end-to-end chemical orientation of an oligonucleotide based on the chemical convention of numbering of carbon atoms in the sugar moiety meaning that there will be a 5′-end defined by the 5′ carbon of the sugar moiety, and a 3′-end defined by the 3′ carbon of the sugar moiety. In a duplex or double stranded oligonucleotide, the respective strands run in opposite 5′ to 3′ directions to permit base pairing between them.
As used herein, “duplex” or also abbreviated as “dup” means two or more complementary strand regions, or strands, of an oligonucleotide or oligonucleotides, hybridized together by way of non-covalent, sequence-specific interaction therebetween. Most commonly, the hybridization in the duplex will be between nucleobases adenine (A) and thymine (T), and/or (A) adenine and uracil (U), and/or guanine (G) and cytosine (C). The duplex may be part of a single stranded structure, wherein self-complementarity leads to hybridization, or as a result of hybridization between respective strands in a double stranded construct.
As used herein, “double strand” or “double stranded” means a pair of oligomeric compounds that are hybridized to one another. In certain embodiments, a double-stranded oligomeric compound comprises a first and a second oligomeric compound.
As used herein, “expression” means the process by which a gene ultimately results in a protein. Expression includes, but is not limited to, transcription, post-transcriptional modification (e.g., splicing, polyadenlyation, addition of 5′-cap), and translation.
As used herein, “transcription” or “transcribed” means the first of several steps of DNA based gene expression in which a target sequence of DNA is copied into RNA (especially mRNA) by the enzyme RNA polymerase. During transcription, a DNA sequence is read by an RNA polymerase, which produces a complementary, antiparallel RNA sequence called a primary transcript.
As used herein, “target sequence” means a sequence to which an oligomeric compound is intended to hybridize to result in a desired activity with respect to APOC3 expression. Oligonucleotides have sufficient complementarity to their target sequences to allow hybridization under physiological conditions.
As used herein, “nucleobase complementarity” or “complementarity” when in reference to nucleobases means a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In both DNA and RNA, guanine (G) is complementary to cytosine (C). In certain embodiments, complementary nucleobase means a nucleobase of an oligomeric compound that is capable of base pairing with a nucleobase of its target sequence. For example, if a nucleobase at a certain position of an oligomeric compound is capable of hydrogen bonding with a nucleobase at a certain position of a target sequence, then the position of hydrogen bonding between the oligomeric compound and the target sequence is considered to be complementary at that nucleobase pair. Nucleobases comprising certain modifications may maintain the ability to pair with a counterpart nucleobase and thus, are still capable of nucleobase complementarity.
As used herein, “non-complementary” in reference to nucleobases means a pair of nucleobases that do not form hydrogen bonds with one another.
As used herein, “complementary” in reference to oligomeric compounds (e.g., linked nucleosides, oligonucleotides) means the capacity of such oligomeric compounds or regions thereof to hybridize to a target sequence, or to a region of the oligomeric compound itself, through nucleobase complementarity. Complementary oligomeric compounds need not have nucleobase complementarity at each nucleoside.
Rather, some mismatches are tolerated. In certain embodiments, complementary oligomeric compounds or regions are complementary at 70% of the nucleobases (70% complementary). In certain embodiments, complementary oligomeric compounds or regions are 80%>complementary. In certain embodiments, complementary oligomeric compounds or regions are 90%>complementary. In certain embodiments, complementary oligomeric compounds or regions are at least 95% complementary. In certain embodiments, complementary oligomeric compounds or regions are 100% complementary. As used herein, “self-complementarity” in reference to oligomeric compounds means a compound that may fold back on itself, creating a duplex as a result of nucleobase hybridization of internal complementary strand regions. Depending on how close together and/or how long the strand regions are, then the compound may form hairpin loops, junctions, bulges or internal loops.
As used herein, “mismatch” means a nucleobase of an oligomeric compound that is not capable of pairing with a nucleobase at a corresponding position of a target sequence, or at a corresponding position of the oligomeric compound itself when the oligomeric compound hybridizes as a result of self-complementarity, when the oligomeric compound and the target sequence and/or self-complementary regions of the oligomeric compound, are aligned.
As used herein, “hybridization” means the pairing of complementary oligomeric compounds (e.g., an oligomeric compound and its target sequence). While not limited to a particular mechanism, the most common mechanism of pairing involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases.
As used herein, “specifically hybridizes” means the ability of an oligomeric compound to hybridize to one nucleic acid site with greater affinity than it hybridizes to another nucleic acid site.
As used herein, “fully complementary” in reference to an oligomeric compound or region thereof means that each nucleobase of the oligomeric compound or region thereof is capable of pairing with a nucleobase of a complementary nucleic acid target sequence or a self-complementary region of the oligomeric compound. Thus, a fully complementary oligomeric compound or region thereof comprises no mismatches or unhybridized nucleobases with respect to its target sequence or a self-complementary region of the oligomeric compound.
As used herein, “percent complementarity” means the percentage of nucleobases of an oligomeric compound that are complementary to an equal-length portion of a target nucleic acid. Percent complementarity is calculated by dividing the number of nucleobases of the oligomeric compound that are complementary to nucleobases at corresponding positions in the target nucleic acid by the total length of the oligomeric compound.
As used herein, “percent identity” means the number of nucleobases in a first nucleic acid that are the same type (independent of chemical modification) as nucleobases at corresponding positions in a second nucleic acid, divided by the total number of nucleobases in the first nucleic acid.
As used herein, “modulation” means a change of amount or quality of a molecule, function, or activity when compared to the amount or quality of a molecule, function, or activity prior to modulation. For example, modulation includes the change, either an increase (stimulation or induction) or a decrease (inhibition or reduction) in gene expression.
As used herein, “type of modification” in reference to a nucleoside or a nucleoside of a “type” means the chemical modification of a nucleoside and includes modified and unmodified nucleosides. Accordingly, unless otherwise indicated, a “nucleoside having a modification of a first type” may be an unmodified nucleoside.
As used herein, “differently modified” means chemical modifications or chemical substituents that are different from one another, including absence of modifications. Thus, for example, a MOE nucleoside and an unmodified naturally occurring RNA nucleoside are “differently modified,” even though the naturally occurring nucleoside is unmodified. Likewise, DNA and RNA oligonucleotides are “differently modified,” even though both are naturally-occurring unmodified nucleosides. Nucleosides that are the same but for comprising different nucleobases are not differently modified. For example, a nucleoside comprising a 2′—OMe modified sugar moiety and an unmodified adenine nucleobase and a nucleoside comprising a 2′—OMe modified sugar moiety and an unmodified thymine nucleobase are not differently modified.
As used herein, “the same type of modifications” refers to modifications that are the same as one another, including absence of modifications. Thus, for example, two unmodified RNA nucleosides have “the same type of modification,” even though the RNA nucleosides are unmodified. Such nucleosides having the same type modification may comprise different nucleobases.
As used herein, “region” or “regions”, or “portion” or “portions”, mean a plurality of linked nucleosides that have a function or character as defined herein, in particular with reference to the claims and definitions as provided herein. Typically such regions or portions comprise at least 10, at least 11, at least 12 or at least 13 linked nucleosides. For example, such regions can comprise 13 to 20 linked nucleosides, such as 13 to 16 or 18 to 20 linked nucleosides. Typically a first region as defined herein consists essentially of 18 to 20 nucleosides and a second region as defined herein consists essentially of 13 to 16 linked nucleosides.
As used herein, “pharmaceutically acceptable carrier or diluent” means any substance suitable for use in administering to an animal. In certain embodiments, a pharmaceutically acceptable carrier or diluent is sterile saline. In certain embodiments, such sterile saline is pharmaceutical grade saline.
As used herein, “substituent” and “substituent group,” means an atom or group that replaces the atom or group of a named parent compound. For example a substituent of a modified nucleoside is any atom or group that differs from the atom or group found in a naturally occurring nucleoside (e.g., a modified 2′-substituent is any atom or group at the 2′-position of a nucleoside other than H or OH). Substituent groups can be protected or unprotected. In certain embodiments, compounds of the present disclosure have substituents at one or at more than one position of the parent compound. Substituents may also be further substituted with other substituent groups and may be attached directly or via a linking group such as oxygen or an alkyl or hydrocarbyl group to a parent compound.
Such substituents can be present as the modification on the sugar moiety, in particular a substituent present at the 2′-position of the sugar moiety. Unless otherwise indicated, groups amenable for use as substituents include without limitation, one or more of halo, hydroxyl, alkyl, alkenyl, alkynyl, acyl, carboxyl, alkoxy, alkoxyalkylene and amino substituents. Certain substituents as described herein can represent modifications directly attached to a ring of a sugar moiety (such as a halo, such as fluoro, directly attached to a sugar ring), or a modification indirectly linked to a ring of a sugar moiety by way of an oxygen linking atom that itself is directly linked to the sugar moiety (such as an alkoxyalkylene, such as methoxyethylene, linked to an oxygen atom, overall providing an MOE substituent as described herein attached to the 2′-position of the sugar moiety).
As used herein, “alkyl,” as used herein, means a saturated straight or branched monovalent C1-6 hydrocarbon radical, with methyl being a most preferred alkyl as a substituent at the 2′-position of the sugar moiety. The alkyl group typically attaches to an oxygen linking atom at the 2′position of the sugar, therefore, overall providing a—Oalkyl substituent, such as an —OCH3 substituent, on a sugar moiety of an oligomeric compound as described herein. This will be well understood be a person skilled in the art.
As used herein, “alkylene” means a saturated straight or branched divalent hydrocarbon radical of the general formula —CnH2n— where n is 1-6. Methylene or ethylene are preferred alkylenes.
As used herein, “alkenyl” means a straight or branched unsaturated monovalent C2-6 hydrocarbon radical, with ethenyl or propenyl being most preferred alkenyls as a substituent at the 2′-position of the sugar moiety. As will be well understood in the art, the degree of unsaturation that is present in an alkenyl radical is the presence of at least one carbon to carbon double bond. The alkenyl group typically attaches to an oxygen linking atom at the 2′-position of the sugar, therefore, overall providing a—Oalkenyl substituent, such as an —OCH2CH═CH2 substituent, on a sugar moiety of an oligomeric compound as described herein. This will be well understood be a person skilled in the art.
As used herein, “alkynyl” means a straight or branched unsaturated C2-6 hydrocarbon radical, with ethynyl being a most preferred alkynyl as a substituent at the 2′-position of the sugar moiety. As will be well understood in the art, the degree of unsaturation that is present in an alkynyl radical is the presence of at least one carbon to carbon triple bond. The alkynyl group typically attaches to an oxygen linking atom at the 2′-position of the sugar, therefore, overall providing a—Oalkynyl substituent on a sugar moiety of an oligomeric compound as described herein. This will be well understood be a person skilled in the art.
As used herein, “carboxyl” is a radical having a general formula —CO2H.
As used herein, “acyl” means a radical formed by removal of a hydroxyl group from a carboxyl radical as defined herein and has the general Formula —C(O)—X where X is typically Cis alkyl.
As used herein, “alkoxy” means a radical formed between an alkyl group, such as a C1-6 alkyl group, and an oxygen atom wherein the oxygen atom is used to attach the alkoxy group either to a parent molecule (such as at the 2′-position of a sugar moiety), or to another group such as an alkylene group as defined herein. Examples of alkoxy groups include without limitation, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy and tert-butoxy. Alkoxy groups as used herein may optionally include further substituent groups.
As used herein, alkoxyalkylene means an alkoxy group as defined herein that is attached to an alkylene group also as defined herein, and wherein the oxygen atom of the alkoxy group attaches to the alkylene group and the alkylene attaches to a parent molecule. The alkylene group typically attaches to an oxygen linking atom at the 2′-position of the sugar, therefore, overall providing a—Oalkylenealkoxy substituent, such as an —OCH2CH2OCH3 substituent, on a sugar moiety of an oligomeric compound as described herein. This will be well understood by a person skilled in the art and is generally referred to as an MOE substituent as defined herein and as known in the art.
As used herein, “amino” includes primary, secondary and tertiary amino groups.
As used herein, “halo” and “halogen,” mean an atom selected from fluorine, chlorine, bromine and iodine.
As used herein, the term “mxRNA” is in particular understood as defined in WO 2020/044186 A2 which is incorporated by reference herein in its entirety.
It will also be understood that oligomeric compounds as described herein may have one or more non-hybridizing nucleosides at one or both ends of one or both strands (overhangs) and/or one or more internal non-hybridizing nucleosides (mismatches) provided there is sufficient complementarity to maintain hybridization under physiologically relevant conditions. Alternatively, oligomeric compounds as described herein may be blunt ended at at least one end.
The term “comprising” is used herein to mean including the method steps or elements identified, but that such steps or elements do not comprise an exclusive list and as such there may be present additional steps or elements.
Further, to the extent that the term “includes” is used in either the detailed description or the claims, such term is intended to be inclusive in a manner similar to the term “comprising” as “comprising” is interpreted when employed as a transitional word in a claim.
The following exemplary embodiments (items) are provided:

    
    
        1. An oligomeric compound capable of inhibiting expression of APOC3, wherein the compound comprises at least a first region of linked nucleosides having at least a first nucleobase sequence that is at least partially complementary to at least a portion of RNA transcribed from an APOC3 gene, wherein the first nucleobase sequence is selected from the following sequences, or a portion thereof: sequences of Tables 1a and 2a (SEQ ID NOs: 1 to 391), wherein the portion preferably has a length of at least 18 nucleotides.
    
    


Said first region is also referred to as the antisense region, and said second region is also referred to as the sense region. As disclosed in preferred embodiments below, said two regions may be located on the same strand, preferably in an adjacent manner. This gives rise to hairpin molecules, also referred to as mxRNAs. On the other hand, said two regions may be located on separate strands which gives rise to double-stranded RNAs (dsRNAs), wherein preferably each strand consists of the respective region. Moreover, said regions may serve as building blocks for muRNAs (see above at Aspect 1). In other words, said first and said second region as defined herein may be used, in accordance with the following definition of muRNAs as first and third regions, respectively:
A nucleic acid construct (muRNA) comprising at least:

    
    
        (a) a first nucleic acid portion that is at least partially complementary to at least a first portion of an RNA which is transcribed from a APOC3 gene;
        (b) a second nucleic acid portion that is at least partially complementary to at least a second portion of an RNA which is transcribed from another gene;
        (c) a third nucleic acid portion that is at least partially complementary to said first nucleic acid portion of (a), so as to form a first nucleic acid duplex region therewith; and
        (d) a fourth nucleic acid portion that is at least partially complementary to said second nucleic acid portion of (b), so as to form a second nucleic acid duplex region therewith.
    
    


Preferred embodiments of and further aspects relating to muRNAs are disclosed in WO2020/065602.

    
    
        2. The oligomeric compound according to item 1, which further comprises at least a second region of linked nucleosides having at least a second nucleobase sequence that is at least partially complementary to the first nucleobase sequence and is selected from the following sequences, or a portion thereof: sequences of Tables 1c and 2c (SEQ ID NOs: 401 to 791), wherein the portion preferably has a length of at least 11 nucleotides, or wherein the portion preferably has a length of at least 8, 9, 10 or 11 nucleotides, more preferably at least 10 nucleotides.
        3. The oligomeric compound according to item 1 or 2, wherein the first nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 175, 293, 262, 297, 277, 366, 337, 254, 274, 286, 137, 149, 280, 343, 225, 221, 185, 373, 121, 281, 331, 367, 296, 28, 345, 328, 339, 278, 271, 212, 223, 369, 276, 332, 300, 341, 334, 138, 193, 340, 31, 167, 275, 191, 336, 90, 346, 219, 283, 213, 23, 24, 285, 347, 370, 206, 282, 342, 272, 303, 220, 209, 29, 89, 291, 117, 372, 218, 368, 148, 217, 128, 338, 171, 94, 324, and 299.
        4. The oligomeric compound according to item 3, wherein the second nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 575, 693, 662, 697, 677, 766, 737, 654, 674, 686, 537, 549, 680, 743, 625, 621, 585, 773, 521, 681, 731, 767, 696, 428, 745, 728, 739, 678, 671, 612, 623, 769, 676, 732, 700, 741, 734, 538, 593, 740, 431, 567, 675, 591, 736, 490, 746, 619, 683, 613, 423, 424, 685, 747, 770, 606, 682, 742, 672, 703, 620, 609, 429, 489, 691, 517, 772, 618, 768, 548, 617, 528, 738, 571, 494, 724, and 699.
        5. The oligomeric compound according to any of items 1 to 4, wherein the first nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 277, 337, 28, 343, 369, 366, 274, 367, 336, 332, 293, 373, 280, 221, 334, 286, 149, 193, 328, 175, 262, 254, 185, 328, 271, 137, 225, 167, 297, and 191.
        6. The oligomeric compound according to item 5, wherein the second nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 677, 737, 428, 743, 769, 766, 674, 767, 736, 732, 693, 773, 680, 621, 734, 686, 549, 593, 728, 575, 662, 654, 585, 728, 671, 537, 625, 567, 697, and 591.
        7. The oligomeric compound according to any of items 1 to 6, wherein the first nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 28, 277, 336, 337, 366, 367, and 369, preferably SEQ ID NO: 28 or 277, more preferably SEQ ID NO: 28.
    
    


These embodiments define antisense nucleobase sequences which provide for surprisingly outstanding performance. For evidence, reference is made to the Examples.

    
    
        8. The oligomeric compound according to item 7, wherein the second nucleobase sequence is selected from the following sequences, or a portion thereof: SEQ ID NOs: 428, 677, 736, 737, 766, 767, and 769, preferably SEQ ID NO: 428 or 677, more preferably SEQ ID NO: 428.
        9. The oligomeric compound according to any of items 1 to 8, wherein the first region of linked nucleosides consists essentially of 18 to 35, preferably 18 to 20, more preferably 18 or 19, and yet more preferably 19 linked nucleosides.
        10. The oligomeric compound according to any of items 2 to 9, wherein the second region of linked nucleosides consists essentially of 11 to 35, preferably 11 to 20, more preferably 13 to 16, and yet more preferably 14 or 15, most preferably 14 linked nucleosides; or wherein the second region of linked nucleosides consists essentially of 10 to 35, preferably 10 to 20, more preferably 10 to 16, and yet more preferably 10 to 15 linked nucleosides.
        11. The oligomeric compound according to any of items 2 to 10, which comprises at least one complementary duplex region that comprises at least a portion of the first nucleoside region directly or indirectly linked to at least a portion of the second nucleoside region, wherein preferably the duplex region has a length of 11 to 19, more preferably 14 to 19, and yet more preferably 14 or 15 base pairs, most preferably 14 base pairs, wherein optionally there is one mismatch within the duplex region; or which comprises at least one complementary duplex region that comprises at least a portion of the first nucleoside region directly or indirectly linked to at least a portion of the second nucleoside region, wherein preferably the duplex region has a length of 10 to 19, more preferably 12 to 19, and yet more preferably 12 to 15 base pairs, wherein optionally there is one mismatch within the duplex region.
        12. The oligomeric compound according to item 11, wherein each of the first and second nucleoside regions has a 5′ to 3′ directionality thereby defining 5′ and 3′ regions respectively thereof.
        13. The oligomeric compound according to item 12, wherein the 5′ region of the first nucleoside region is directly or indirectly linked to the 3′ region of the second nucleoside region, for example by complementary base pairing, and/or wherein the 3′ region of the first nucleoside region is directly or indirectly linked to the 5′ region of the second nucleoside region, wherein preferably the 5′ terminal nucleoside of the first nucleoside region base pairs with the 3′ terminal nucleoside of the second nucleoside region; or wherein the 5′ region of the first nucleoside region is directly or indirectly linked to the 3′ region of the second nucleoside region, for example by complementary base pairing, wherein preferably the 5′ terminal nucleoside of the first nucleoside region base pairs with the 3′ terminal nucleoside of the second nucleoside region.
        14. The oligomeric compound according to item 12 or 13, wherein the 3′ region of the first nucleoside region is directly or indirectly linked to the 5′ region of the second nucleoside region, wherein preferably the first nucleoside region is directly and covalently linked to the second nucleoside region such as by a phosphate, a phosphorothioate, or a phosphorodithoate.
        15. The oligomeric compound according to any of items 1 to 14, which further comprises one or more ligands.
        16. The oligomeric compound according to item 15, wherein the one or more ligands are conjugated to the second nucleoside region and/or the first nucleoside region.
        17. The oligomeric compound according to item 16, as dependent on claim 12, wherein the one or more ligands are conjugated at the 3′ region, preferably to the 3′ end of the second nucleoside region and/or of the first nucleoside region, and/or to the 5′ end of the second nucleoside region.
        18. The oligomeric compound according to any of item 15 to 17, wherein the one or more ligands are any cell directing moiety, such as lipids, carbohydrates, aptamers, vitamins and/or peptides that bind cellular membrane or a specific target on cellular surface.
        19. The oligomeric compound according to item 18, wherein the one or more ligands comprise one or more carbohydrates.
        20. The oligomeric compound according to item 19, wherein the one or more carbohydrates can be a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide or polysaccharide.
        21. The oligomeric compound according to item 20, wherein the one or more carbohydrates comprise or consist of one or more hexose moieties.
        22. The oligomeric compound according to item 21, wherein the one or more hexose moieties are one or more galactose moieties, one or more lactose moieties, one or more N-Acetyl-Galactosamine moieties, and/or one or more mannose moieties.
        23. The oligomeric compound according to item 22, wherein the one or more carbohydrates comprise one or more N-Acetyl-Galactosamine moieties.
        24. The oligomeric compound according to item 23, which comprises two or three N-Acetyl-Galactosamine moieties, preferably three.
        25. The oligomeric compound according to any of items 15 to 24, wherein the one or more ligands are attached to the oligomeric compound, preferably to the second nucleoside region thereof, in a linear configuration, or in a branched configuration.
        26. The oligomeric compound according to item 25, wherein the one or more ligands are attached to the oligomeric compound as a biantennary or triantennary configuration.
        27. The oligomeric compound according to any one of items 1 to 26, wherein the compound consists of the first region of linked nucleosides and the second region of linked nucleosides.
    
    


Each of said regions may constitute a separate strand, thereby giving rise to a double-stranded RNA (dsRNA). Particularly preferred dsRNAs are those with a length of the first strand of 19 nucleosides and a length of the second region of 14 or 15, preferably 14 nucleosides. When used for defining the length of a region or strand, the terms “nucleoside” and “nucleotide” (sometimes abbreviated “nt”) are used equivalently.

    
    
        28. The oligomeric compound according to item 12, wherein the oligomeric compound comprises a single strand comprising the first and second nucleoside regions, wherein the single strand dimerises whereby at least a portion of the first nucleoside region is directly or indirectly linked to at least a portion of the second nucleoside region so as to form the at least partially complementary duplex region.
    
    


In other words, the oligomeric compound comprises a single strand comprising the first and second nucleoside regions, wherein at least a portion of the first nucleoside region is directly or indirectly linked to at least a portion of the second nucleoside region so as to form the at least partially complementary duplex region.

    
    
        29. The oligomeric compound according to item 28, wherein the first nucleoside region has a greater number of linked nucleosides compared to the second nucleoside region, whereby the additional number of linked nucleosides of the first nucleoside region form a hairpin loop linking the first and second nucleoside regions.
    
    


Such compounds are also referred to as hairpins or mxRNAs herein.

    
    
        30. The oligomeric compound according to item 29, as dependent on claim 12, whereby the hairpin loop is present at the 3′ region of the first nucleoside region.
        31. The oligomeric compound according to item 29 or 30, wherein the hairpin loop comprises 4 or 5 linked nucleosides.
    
    


Particularly advantageous is a length of the first region of 19 nucleosides, of the second region of 14 nucleotides, and of the hairpin loop of five nucleotides, wherein the five nucleotides in the hairpin are the five 3′-terminal nucleosides of the first region. Such molecular architecture of a hairpin or mxRNA is also designated “14-5-14” herein.

    
    
        32. The oligomeric compound according to any one of items 28 to 31, wherein the single strand has a nucleobase sequence selected from SEQ ID NOs: 792 to 803, preferably from SEQ ID NOs: 792, 793, 796, 800 and 803, most preferably from SEQ ID NOs: 796 and 803 particularly SEQ ID NO: 803.
        33. The oligomeric compound according to item 32, wherein the single strand is selected from Table 3b, in particular from constructs A28(14-4)mF and A277(12-5_, A28(14-4)mF being especially advantageous.
        34. The oligomeric compound according to any of items 1 to 33, which comprises internucleoside linkages and wherein at least one internucleoside linkage is a modified internucleoside linkage.
    
    


Specific modified internucleoside linkages are the subject of the embodiments which follow. Certain modified internucleoside linkages are known in the art and described in, for example, Hu et al., Signal Transduction and Targeted Therapy (2020)5:101.

    
    
        35. The oligomeric compound according to item 34, wherein the modified internucleoside linkage is a phosphorothioate or phosphorodithioate internucleoside linkage.
        36. The oligomeric compound according to item 35, which comprises 1 to 15 phosphorothioate or phosphorodithioate internucleoside linkages.
        37. The oligomeric compound according to item 36, which comprises 7, 8, 9 or 10 phosphorothioate or phosphorodithioate internucleoside linkages.
        38. The oligomeric compound according to any of items 35 to 37, as dependent on item 12, which comprises one or more phosphorothioate or phosphorodithioate internucleoside linkages at the 5′ region of the first nucleoside region.
        39. The oligomeric compound according to any of items 35 to 38, as dependent on item 12, which comprises one or more phosphorothioate or phosphorodithioate internucleoside linkages at the 5′ region of the second nucleoside region.
        40. The oligomeric compound according to any of items 35 to 39, as dependent on item 28, which comprises phosphorothioate or phosphorodithioate internucleoside linkages between at least two, preferably at least three, preferably at least four, preferably at least five, adjacent nucleosides of the hairpin loop, dependent on the number of nucleotides present in the hairpin loop.
        41. The oligomeric compound according to item 40, which comprises a phosphorothioate or phosphorodithioate internucleoside linkage between each adjacent nucleoside that is present in the hairpin loop.
        42. The oligomeric compound according to any of items 1 to 41, wherein at least one nucleoside comprises a modified sugar.
    
    


Preferred modified sugars are subject of the embodiments which follow. Certain modified sugars are known in the art and described in, for example, Hu et al., Signal Transduction and Targeted Therapy (2020)5:101.

    
    
        43. The oligomeric compound according to item 42, wherein the modified sugar is selected from 2′ modified sugars, locked nucleic acid (LNA) sugar, (S)—constrained ethyl bicyclic nucleic acid sugar, tricyclo-DNA sugar, morpholino, unlocked nucleic acid (UNA) sugar, and glycol nucleic acid (GNA) sugar.
        44. The oligomeric compound according to item 43, wherein the 2′ modified sugar is selected from 2′-O-methyl modified sugar, 2′-O-methoxyethyl modified sugar, 2′-F modified sugar, 2′-arabino-fluoro modified sugar, 2′-O-benzyl modified sugar, and 2′-O-methyl-4-pyridine modified sugar.
        45. The oligomeric compound according to item 44, wherein at least one modified sugar is a 2′-O-methyl modified sugar.
        46. The oligomeric compound according to item 44 or 45, wherein at least one modified sugar is a 2′-F modified sugar.
        47. The oligomeric compound of item 45 or 46, wherein the sugar is ribose.
        48. The oligomeric compound according to any of items 45 to 48, as dependent on item 12, wherein sugars of the nucleosides at any of positions 2 and 14 downstream from the first nucleoside of the 5′ region of the first nucleoside region, do not contain 2′-O-methyl modifications.
        49. The oligomeric compound according to any of items 45 to 48, as dependent on item 12, wherein sugars of the nucleosides of the second nucleoside region, that correspond in position to any of the nucleosides of the first nucleoside region at any of positions 9 to 11 downstream from the first nucleotide of the 5′ region of the first nucleoside region, in particular from sequence A277(12-5) and A28(14-4)mF do not contain 2′-O-methyl modifications.
        50. The oligomeric compound of any one of items 45 to 49, wherein the 3′ terminal position of the second nucleoside region does not contain a 2′-O-methyl modification.
        51. The oligomeric compound according to item 49 or 50, wherein sugars of the nucleosides at any of positions 2 and 14 downstream from the first nucleoside of the 5′ region of the first nucleoside region, contain 2′-F modifications.
        52. The oligomeric compound according to any of items 49 to 51, wherein sugars of the nucleosides of the second nucleoside region, that correspond in position to any of the nucleosides of the first nucleoside region at any of positions 9 to 11 downstream from the first nucleoside of the 5′ region of the first nucleoside region, contain 2′-F modifications.
        53. The oligomeric compound of item 51 or 52, wherein the 3′ terminal position of the second nucleoside region contains a 2′-F modification.
        54. The oligomeric compound according to any of items 47 to 53, as dependent on item 12, wherein one or more of the odd numbered nucleosides starting from the 5′ region of the first nucleoside region are modified, and/or wherein one or more of the even numbered nucleotides starting from the 5′ region of the first nucleoside region are modified, wherein typically the modification of the even numbered nucleotides is a second modification that is different from the modification of odd numbered nucleotides.
        55. The oligomeric compound according to item 54, wherein one or more of the odd numbered nucleosides starting from the 3′ region of the second nucleoside region are modified by a modification that is different from the modification of odd numbered nucleosides of the first nucleoside region.
        56. The oligomeric compound according to item 54 or 55, wherein one or more of the even numbered nucleosides starting from the 3′ region of the second nucleoside region are modified by a modification that is different from the modification of even numbered nucleosides of the first nucleoside region according to item 55.
        57. The oligomeric compound according to any of items 54 to 56, wherein at least one or more of the modified even numbered nucleosides of the first nucleoside region is adjacent to at least one or more of the differently modified odd numbered nucleosides of the first nucleoside region.
        58. The oligomeric compound according to any of items 54 to 57, wherein at least one or more of the modified even numbered nucleosides of the second nucleoside region is adjacent to at least one or more of the differently modified odd numbered nucleosides of the second nucleoside region.
        59. The oligomeric compound according to any of items 54 to 58, wherein sugars of one or more of the odd numbered nucleosides starting from the 5′ region of the first nucleoside region are 2′-O-methyl modified sugars.
        60. The oligomeric compound according to any of items 54 to 59, wherein one or more of the even numbered nucleosides starting from the 5′ region of the first nucleoside region are 2′-F modified sugars.
        61. The oligomeric compound according to any of items 54 to 60, wherein sugars of one or more of the odd numbered nucleosides starting from the 3′ region of the second nucleoside region are 2′-F modified sugars.
        62. The oligomeric compound according to any of items 54 to 61, wherein one or more of the even numbered nucleosides starting from the 3′ region of the second nucleoside region are 2′-O-methyl modified sugars.
        63. The oligomeric compound according to any of items 42 to 62, wherein sugars of a plurality of adjacent nucleosides of the first nucleoside region are modified by a common modification.
        64. The oligomeric compound according to any of items 42 to 63, wherein sugars of a plurality of adjacent nucleosides of the second nucleoside region are modified by a common modification.
        65. The oligomeric compound according to any of items 54 to 64, as dependent on item 31, wherein sugars of a plurality of adjacent nucleosides of the hairpin loop are modified by a common modification.
        66. The oligomeric compound according to any of items 63 to 65, wherein the common modification is a 2′-F modified sugar.
        67. The oligomeric compound according to any of items 63 to 65, wherein the common modification is a 2′-O-methyl modified sugar.
        68. The oligomeric compound according to item 67, wherein the plurality of adjacent 2′-O-methyl modified sugars are present in at least eight adjacent nucleosides of the first and/or second nucleoside regions.
        69. The oligomeric compound according to item 67, wherein the plurality of adjacent 2′-O-methyl modified sugars are present in three or four adjacent nucleosides of the hairpin loop.
        70. The oligomeric compound according to item 42, as dependent on item 29, wherein the hairpin loop comprises at least one nucleoside having a modified sugar.
        71. The oligomeric compound according to item 70, wherein the at least one nucleoside is adjacent a nucleoside with a differently modified sugar.
        72. The oligomeric compound according to item 71, wherein the modified sugar is a 2′-O-methyl modified sugar, and the differently modifies sugar is a 2′-F modified sugar.
        73. The oligomeric compound according to any of items 1 to 72, which comprises one or more nucleosides having an un-modified sugar moiety.
        74. The oligomeric compound according to item 73, wherein the unmodified sugar is present in the 5′ region of the second nucleoside region.
        75. The oligomeric compound according to item 73 or 74, as dependent on item 29, wherein the unmodified sugar is present in the hairpin loop.
        76. The oligomeric compound according to any of items 1 to 75, wherein one or more nucleosides of the first nucleoside region and/or the second nucleoside region is an inverted nucleoside and is attached to an adjacent nucleoside via the 3′ carbon of its sugar and the 3′ carbon of the sugar of the adjacent nucleoside, and/or one or more nucleosides of the first nucleoside region and/or the second nucleoside region is an inverted nucleoside and is attached to an adjacent nucleoside via the 5′ carbon of its sugar and the 5′ carbon of the sugar of the adjacent nucleoside.
        77. The oligomeric compound according to any of items 1 to 76, which is blunt ended.
        78. The oligomeric compound according to any of items 1 to 76, wherein either the first or second nucleoside region has an overhang.
        79. The oligomeric compound according to any one of the preceding items, wherein the first region of linked nucleotides is selected from Table 1 b or Table 2b, preferably from the entries in Table 1 b which have a nucleobase sequence as defined in any one of item 3, 5 or 7.
        80. The oligomeric compound according to any one of the preceding items, wherein the second region of linked nucleotides is selected from Table 1d or Table 2d, preferably from the entries in Table 1b which have a nucleobase sequence as defined in any one of items 4, 6 or 8.
        81. A composition comprising an oligomeric compound according to any of items 1 to 80, and a physiologically acceptable excipient.
        82. A pharmaceutical composition comprising an oligomeric compound according to any of items 1 to 80.
        83. The pharmaceutical composition of item 82, further comprising a pharmaceutically acceptable excipient, diluent, antioxidant, and/or preservative.
        84. The pharmaceutical composition of item 82 or 83, wherein the oligomeric compound is the only pharmaceutically active agent.
        85. The pharmaceutical composition of item 84, wherein the pharmaceutical composition is to be administered to patients or individuals which are statin-intolerant and/or for whom statins are contraindicated.
        86. The pharmaceutical composition of item 82 or 83, wherein the pharmaceutical composition furthermore comprises one or more further pharmaceutically active agents.
    
    


87. The pharmaceutical composition of item 86, wherein the further pharmaceutically active agent(s) is/are a further oligomeric compound which is directed to a target different from APOC3, preferably PCSK9; Vascepa; Vupanorsen; statins such as Rosuvastatin and Simvastatin; fibrates such fenofibrate; and/or LDL-cholesterol lowering compounds such as statins and ezetimibe.

    
    
        88. The pharmaceutical composition of item 86 or 87, wherein the oligomeric compound and the further pharmaceutically active agent(s) are to be administered concomitantly or in any order.
        89. An oligomeric compound according to any of item 1 to 80, for use in human or veterinary medicine or therapy.
        90. An oligomeric compound according to any of items 1 to 80, for use in a method of treating, ameliorating and/or preventing a disease or disorder.
        91. The compound for use of item 90, wherein the disease or disorder is an APOC3-associated disease or disorder, or a disease or disorder requiring reduction of APOC3 expression levels, the disease or disorder preferably being selected from dyslipidemia including mixed dyslipidemia; hyperchylomicronemia including familial hyperchylomicronemia; hypertriglyceridemia, preferably severe hypertriglyceridemia and/or hypertriglyceridemia with blood triglyceride levels above 500 mg/dl;
    
    


inflammation including low-grade inflammation; atherosclerosis; atherosclerotic cardiovascular diseases (ASCVD) including major adverse cardiovascular events (MACE) such as myocardial infarction, stroke and peripheral arterial disease; and pancreatitis including acute pancreatitis.

    
    
        92. A method of treating a disease or disorder comprising administration of an oligomeric compound according to any of item 1 to 80, to an individual in need of treatment.
        93. The method according to item 92, wherein the oligomeric compound is administered subcutaneously or intravenously to the individual.
        93. Use of an oligomeric compound according to any of item 1 to 80, for use in research as a gene function analysis tool.
        94. Use of an oligomeric compound according to any of items 1 to 80 in the manufacture of a medicament for a treatment of a disease or disorder. The diseases and disorders are preferably the same as set forth under item 91 above.

Effects Achieved by the Oligomeric Compounds

    
    


Due to the use of the oligomeric compounds as described herein, a significant reduction of APOC3 mRNA, especially in vitro or in liver tissues consisting essentially of human hepatocytes, can be achieved as e.g. shown in the examples disclosed herein. In addition, a significant reduction of APCO3 proteins in the plasma level, e.g. of mice having a liver consisting essentially of human hepatocytes, can be achieved by using the oligomeric constructs as described herein. In particular these effects can last over an extended time period such as six weeks, e.g. in mice having a liver consisting essentially of human hepatocytes.
In addition, by using oligomeric compounds as described herein, significant degrees of reduction of triglyceride levels in the serum, in particular of mice having a liver essentially consisting of human hepatocytes, can be achieved, also over an extended period of time, such as six weeks. An unexpected and surprising finding is that, in addition to the reduction of triglycerides in the serum, in particular of the same mice, a significant reduction in the level of cholesterol in the serum is achieved at the same time over an extended time period, such as six weeks.
It has also been surprisingly found that, in certain embodiments, the aforementioned beneficial effects can be achieved by using oligomeric compounds as described herein in the form of shRNA constructs having a reduced number of fluorine substitutions, such as five fluorine substitutions in total, on the respective 2′ positions of their ribose units compared to conventional shRNA molecules having an alternating series of 2′-fluoro and 2′-O-methyl modifications.
Furthermore, it was surprisingly found that, in certain embodiments, the mentioned effects are achieved by using oligomeric compounds as described herein in the form of shRNA constructs as described herein having a reduced length of e.g. 29 linked nucleosides compared to conventional shRNA molecules. The same effects can also surprisingly be achieved for such constructs having a length of the sense strand of about 10 nucleosides.
The aforementioned effects can be achieved by using a dosage of about 10 mg/kg body weight to 30 mg/kg body weight, in particular with respect to mice.
Constructs of the Oligomeric Compounds
The following Tables show nucleobase sequences of antisense and sense strands of oligomeric compounds as described herein, and definitions of antisense and sense strands of modified oligomeric compounds (the notation including nucleobase sequence, sugar modifications, and, where applicable, modified phosphates).
The notation used is common in the art and as the following meaning:

    
    
        A represents adenine;
        U represents uracil;
        C represents cytosine;
        G represents guanine.
        P represents a terminal phosphate group which is preferred but not indispensable;
        m represents a methyl modification at the 2′ position of the sugar of the underlying nucleoside;
        f represents a fluoro modification at the 2′ position of the sugar of the underlying nucleoside.
        r indicates an unmodified (2′-OH) ribonucleotide;
        (ps) or #represents a phosphorothioate inter-nucleoside linkage;
        i represents an inverted inter-nucleoside linkage, which can be either 3′-3′, or 5′-5′;
        vp represents vinyl phosphonate;
        mvp represents methyl vinyl phosphonate;
        3xGaINAc represents a trivalent GaINAc.
    
    


Sometimes, nucleosides are shown in square brackets for ease of reading. This notation does not indicate structural elements or modifications.
To the extent displayed, the presence of a 5′-terminal phosphate (“P”) is optional. Conversely, to the extent a 5′-terminal phosphate is not displayed, its presence is optional as well. Generally, there is no requirement for a 5′-terminal phosphate in compounds to be administered to mammalian cells, since a mammalian kinase adds a 5′-terminal phosphate in the case of its absence.
Furthermore when a notation like “A277(12-5)mF” is used, the term “A277” designates the sequence suitable for RNAi with APOC3, wherein the first number in the round brackets, i.e. 12 in the present case, designates the number of base pairs within a duplex region within a shRNA, and the second number in the round brackets, in this case 5, designates the number of nucleotides present in the hairpin loop of the shRNA. If there is no designation after the hyphen in the round brackets, it means that the loop consists of 5 nucleotides.
Tables 1a to 1d below show nucleobase sequences and sugar-phosphate backbone modifications of antisense and sense strands of the 376 constructs selected in accordance with the Examples. The above disclosed 30 preferred oligomeric compounds have been selected from these 376 constructs. The numbering in Table 1a coincides with the number of the corresponding entry in the sequence listing. For Table 1c the following applies: entry number in the sequence listing=entry number in the Table+400.







TABLE 1a







Nucleobase sequences of the antisense


strands of 376 exemplary constructs








SEQ ID



NO:
Nucleobase sequence











1
UUCUAGGGAUGAACUGAGC


 


2
UCUCUAGGGAUGAACUGAG


 


3
UCCUCUAGGGAUGAACUGA


 


4
UGCCUCUAGGGAUGAACUG


 


5
UUGCCUCUAGGGAUGAACU


 


6
UCUGCCUCUAGGGAUGAAC


 


7
UGCUGCCUCUAGGGAUGAA


 


8
UAGCUGCCUCUAGGGAUGA


 


9
UGCAGCUGCCUCUAGGGAU


 


10
UAGCAGCUGCCUCUAGGGA


 


11
UGAGCAGCUGCCUCUAGGG


 


12
UGGAGCAGCUGCCUCUAGG


 


13
UUGUUCCUGGAGCAGCUGC


 


14
UCUGUUCCUGGAGCAGCUG


 


15
UCCUCUGUUCCUGGAGCAG


 


16
UACCUCUGUUCCUGGAGCA


 


17
UCACCUCUGUUCCUGGAGC


 


18
UGCACCUCUGUUCCUGGAG


 


19
UGGCACCUCUGUUCCUGGA


 


20
UUGGCACCUCUGUUCCUGG


 


21
UAUGGCACCUCUGUUCCUG


 


22
UCAUGGCACCUCUGUUCCU


 


23
UUGCAUGGCACCUCUGUUC


 


24
UCUGCAUGGCACCUCUGUU


 


25
UGCUGCAUGGCACCUCUGU


 


26
UGGCUGCAUGGCACCUCUG


 


27
UGGGCUGCAUGGCACCUCU


 


28
UCAACAAGGAGUACCCGGG


 


29
UACAACAAGGAGUACCCGG


 


30
UAACAACAAGGAGUACCCG


 


31
UCAACAACAAGGAGUACCC


 


32
UGCAACAACAAGGAGUACC


 


33
UGGCAACAACAAGGAGUAC


 


34
UGGGCAACAACAAGGAGUA


 


35
UAGGGCAACAACAAGGAGU


 


36
UGAGGGCAACAACAAGGAG


 


37
UGGAGGGCAACAACAAGGA


 


38
UAGGAGGGCAACAACAAGG


 


39
UCAGGAGGGCAACAACAAG


 


40
UCCAGGAGGGCAACAACAA


 


41
UGCCAGGAGGGCAACAACA


 


42
UCGCCAGGAGGGCAACAAC


 


43
UGCGCCAGGAGGGCAACAA


 


44
UAGCGCCAGGAGGGCAACA


 


45
UGAGCGCCAGGAGGGCAAC


 


46
UGGAGCGCCAGGAGGGCAA


 


47
UAGGAGCGCCAGGAGGGCA


 


48
UGCCAGGAGCGCCAGGAGG


 


49
UAGAGGCCAGGAGCGCCAG


 


50
UCAGAGGCCAGGAGCGCCA


 


51
UGCAGAGGCCAGGAGCGCC


 


52
UGGCAGAGGCCAGGAGCGC


 


53
UGGGCAGAGGCCAGGAGCG


 


54
UUCGGGCAGAGGCCAGGAG


 


55
UCUCGGGCAGAGGCCAGGA


 


56
UGCUCGGGCAGAGGCCAGG


 


57
UAGCUCGGGCAGAGGCCAG


 


58
UAAGCUCGGGCAGAGGCCA


 


59
UGAAGCUCGGGCAGAGGCC


 


60
UUGAAGCUCGGGCAGAGGC


 


61
UCUGAAGCUCGGGCAGAGG


 


62
UUCUGAAGCUCGGGCAGAG


 


63
UCUCUGAAGCUCGGGCAGA


 


64
UCCUCUGAAGCUCGGGCAG


 


65
UGCCUCUGAAGCUCGGGCA


 


66
UGGCCUCUGAAGCUCGGGC


 


67
UCGGCCUCUGAAGCUCGGG


 


68
UUCGGCCUCUGAAGCUCGG


 


69
UCUCGGCCUCUGAAGCUCG


 


70
UCCUCGGCCUCUGAAGCUC


 


71
UUCCUCGGCCUCUGAAGCU


 


72
UAUCCUCGGCCUCUGAAGC


 


73
UCAUCCUCGGCCUCUGAAG


 


74
UGCAUCCUCGGCCUCUGAA


 


75
UGGCAUCCUCGGCCUCUGA


 


76
UAGGCAUCCUCGGCCUCUG


 


77
UGAGGCAUCCUCGGCCUCU


 


78
UGGAGGCAUCCUCGGCCUC


 


79
UGGGAGGCAUCCUCGGCCU


 


80
UAGGGAGGCAUCCUCGGCC


 


81
UAAGGGAGGCAUCCUCGGC


 


82
UGAAGGGAGGCAUCCUCGG


 


83
UAGAAGGGAGGCAUCCUCG


 


84
UGAGAAGGGAGGCAUCCUC


 


85
UUGAGAAGGGAGGCAUCCU


 


86
UCUGAGAAGGGAGGCAUCC


 


87
UGCUGAGAAGGGAGGCAUC


 


88
UAGCUGAGAAGGGAGGCAU


 


89
UUGAAGCUGAGAAGGGAGG


 


90
UAUGAAGCUGAGAAGGGAG


 


91
UCAUGAAGCUGAGAAGGGA


 


92
UGCAUGAAGCUGAGAAGGG


 


93
UUGCAUGAAGCUGAGAAGG


 


94
UCUGCAUGAAGCUGAGAAG


 


95
UCCUGCAUGAAGCUGAGAA


 


96
UCCCUGCAUGAAGCUGAGA


 


97
UACCCUGCAUGAAGCUGAG


 


98
UAACCCUGCAUGAAGCUGA


 


99
UUAACCCUGCAUGAAGCUG


 


100
UGUAACCCUGCAUGAAGCU


 


101
UUGUAACCCUGCAUGAAGC


 


102
UAUGUAACCCUGCAUGAAG


 


103
UCAUGUAACCCUGCAUGAA


 


104
UUCAUGUAACCCUGCAUGA


 


105
UUUCAUGUAACCCUGCAUG


 


106
UCUUCAUGUAACCCUGCAU


 


107
UGCUUCAUGUAACCCUGCA


 


108
UUGCUUCAUGUAACCCUGC


 


109
UGUGCUUCAUGUAACCCUG


 


110
UCGUGCUUCAUGUAACCCU


 


111
UGCGUGCUUCAUGUAACCC


 


112
UGGCGUGCUUCAUGUAACC


 


113
UUGGCGUGCUUCAUGUAAC


 


114
UGUGGCGUGCUUCAUGUAA


 


115
UGGUGGCGUGCUUCAUGUA


 


116
UUGGUGGCGUGCUUCAUGU


 


117
UUUGGUGGCGUGCUUCAUG


 


118
UCUUGGUGGCGUGCUUCAU


 


119
UUCUUGGUGGCGUGCUUCA


 


120
UGUCUUGGUGGCGUGCUUC


 


121
UGGUCUUGGUGGCGUGCUU


 


122
UCGGUCUUGGUGGCGUGCU


 


123
UGCGGUCUUGGUGGCGUGC


 


124
UGGCGGUCUUGGUGGCGUG


 


125
UUGGCGGUCUUGGUGGCGU


 


126
UUUGGCGGUCUUGGUGGCG


 


127
UCUUGGCGGUCUUGGUGGC


 


128
UCCUUGGCGGUCUUGGUGG


 


129
UUCCUUGGCGGUCUUGGUG


 


130
UAUCCUUGGCGGUCUUGGU


 


131
UCAUCCUUGGCGGUCUUGG


 


132
UGCAUCCUUGGCGGUCUUG


 


133
UUGCAUCCUUGGCGGUCUU


 


134
UGUGCAUCCUUGGCGGUCU


 


135
UAGUGCAUCCUUGGCGGUC


 


136
UCAGUGCAUCCUUGGCGGU


 


137
UUCAGUGCAUCCUUGGCGG


 


138
UCUCAGUGCAUCCUUGGCG


 


139
UGCUCAGUGCAUCCUUGGC


 


140
UUGCUCAGUGCAUCCUUGG


 


141
UCUGCUCAGUGCAUCCUUG


 


142
UGCUGCUCAGUGCAUCCUU


 


143
UCGCUGCUCAGUGCAUCCU


 


144
UACGCUGCUCAGUGCAUCC


 


145
UCACGCUGCUCAGUGCAUC


 


146
UGCACGCUGCUCAGUGCAU


 


147
UUGCACGCUGCUCAGUGCA


 


148
UCUGCACGCUGCUCAGUGC


 


149
UCCUGCACGCUGCUCAGUG


 


150
UUCCUGCACGCUGCUCAGU


 


151
UACUCCUGCACGCUGCUCA


 


152
UGGGACUCCUGCACGCUGC


 


153
UUGGGACUCCUGCACGCUG


 


154
UCUGGGACUCCUGCACGCU


 


155
UCCUGGGACUCCUGCACGC


 


156
UACCUGGGACUCCUGCACG


 


157
UCACCUGGGACUCCUGCAC


 


158
UCCACCUGGGACUCCUGCA


 


159
UGGGCCACCUGGGACUCCU


 


160
UUGGGCCACCUGGGACUCC


 


161
UUGCUGGGCCACCUGGGAC


 


162
UCUGCUGGGCCACCUGGGA


 


163
UGGCCUGCUGGGCCACCUG


 


164
UCCUGGCCUGCUGGGCCAC


 


165
UCCAUCGGUCACCCAGCCC


 


166
UGCCAUCGGUCACCCAGCC


 


167
UAGCCAUCGGUCACCCAGC


 


168
UAAGCCAUCGGUCACCCAG


 


169
UGAAGCCAUCGGUCACCCA


 


170
UUGAAGCCAUCGGUCACCC


 


171
UCUGAAGCCAUCGGUCACC


 


172
UACUGAAGCCAUCGGUCAC


 


173
UAACUGAAGCCAUCGGUCA


 


174
UGAACUGAAGCCAUCGGUC


 


175
UGGAACUGAAGCCAUCGGU


 


176
UGGGAACUGAAGCCAUCGG


 


177
UAGGGAACUGAAGCCAUCG


 


178
UCAGGGAACUGAAGCCAUC


 


179
UUCAGGGAACUGAAGCCAU


 


180
UUUCAGGGAACUGAAGCCA


 


181
UUUUCAGGGAACUGAAGCC


 


182
UCUUUCAGGGAACUGAAGC


 


183
UUCUUUCAGGGAACUGAAG


 


184
UGUCUUUCAGGGAACUGAA


 


185
UAGUCUUUCAGGGAACUGA


 


186
UUAGUCUUUCAGGGAACUG


 


187
UGUAGUCUUUCAGGGAACU


 


188
UAGUAGUCUUUCAGGGAAC


 


189
UCAGUAGUCUUUCAGGGAA


 


190
UCCAGUAGUCUUUCAGGGA


 


191
UUCCAGUAGUCUUUCAGGG


 


192
UCUCCAGUAGUCUUUCAGG


 


193
UGCUCCAGUAGUCUUUCAG


 


194
UUGCUCCAGUAGUCUUUCA


 


195
UGUGCUCCAGUAGUCUUUC


 


196
UGGUGCUCCAGUAGUCUUU


 


197
UCGGUGCUCCAGUAGUCUU


 


198
UACGGUGCUCCAGUAGUCU


 


199
UAACGGUGCUCCAGUAGUC


 


200
UUAACGGUGCUCCAGUAGU


 


201
UUUAACGGUGCUCCAGUAG


 


202
UCUUAACGGUGCUCCAGUA


 


203
UCCUUAACGGUGCUCCAGU


 


204
UUCCUUAACGGUGCUCCAG


 


205
UGUCCUUAACGGUGCUCCA


 


206
UUGUCCUUAACGGUGCUCC


 


207
UUUGUCCUUAACGGUGCUC


 


208
UCUUGUCCUUAACGGUGCU


 


209
UACUUGUCCUUAACGGUGC


 


210
UAACUUGUCCUUAACGGUG


 


211
UGAACUUGUCCUUAACGGU


 


212
UAGAACUUGUCCUUAACGG


 


213
UGAGAACUUGUCCUUAACG


 


214
UAGAGAACUUGUCCUUAAC


 


215
UCAGAGAACUUGUCCUUAA


 


216
UUCAGAGAACUUGUCCUUA


 


217
UCUCAGAGAACUUGUCCUU


 


218
UACUCAGAGAACUUGUCCU


 


219
UAACUCAGAGAACUUGUCC


 


220
UGAACUCAGAGAACUUGUC


 


221
UCAGAACUCAGAGAACUUG


 


222
UCCAGAACUCAGAGAACUU


 


223
UCCCAGAACUCAGAGAACU


 


224
UUCCCAGAACUCAGAGAAC


 


225
UAUCCCAGAACUCAGAGAA


 


226
UAAUCCCAGAACUCAGAGA


 


227
UAAAUCCCAGAACUCAGAG


 


228
UCAAAUCCCAGAACUCAGA


 


229
UCCAAAUCCCAGAACUCAG


 


230
UUCCAAAUCCCAGAACUCA


 


231
UGUCCAAAUCCCAGAACUC


 


232
UGGUCCAAAUCCCAGAACU


 


233
UGGGUCCAAAUCCCAGAAC


 


234
UAGGGUCCAAAUCCCAGAA


 


235
UCAGGGUCCAAAUCCCAGA


 


236
UUCAGGGUCCAAAUCCCAG


 


237
UGACCUCAGGGUCCAAAUC


 


238
UUGACCUCAGGGUCCAAAU


 


239
UCUGACCUCAGGGUCCAAA


 


240
UUCUGACCUCAGGGUCCAA


 


241
UGUCUGACCUCAGGGUCCA


 


242
UGGUCUGACCUCAGGGUCC


 


243
UUGGUCUGACCUCAGGGUC


 


244
UUUGGUCUGACCUCAGGGU


 


245
UGUUGGUCUGACCUCAGGG


 


246
UAGUUGGUCUGACCUCAGG


 


247
UAAGUUGGUCUGACCUCAG


 


248
UGAAGUUGGUCUGACCUCA


 


249
UUGAAGUUGGUCUGACCUC


 


250
UCUGAAGUUGGUCUGACCU


 


251
UGGCUGAAGUUGGUCUGAC


 


252
UCGGCUGAAGUUGGUCUGA


 


253
UACGGCUGAAGUUGGUCUG


 


254
UCACGGCUGAAGUUGGUCU


 


255
UCCACGGCUGAAGUUGGUC


 


256
UGCCACGGCUGAAGUUGGU


 


257
UCAGCCACGGCUGAAGUUG


 


258
UGCAGCCACGGCUGAAGUU


 


259
UGGCAGCCACGGCUGAAGU


 


260
UAGGCAGCCACGGCUGAAG


 


261
UCAGGCAGCCACGGCUGAA


 


262
UUCUCAGGCAGCCACGGCU


 


263
UGUCUCAGGCAGCCACGGC


 


264
UGGUCUCAGGCAGCCACGG


 


265
UAGGUCUCAGGCAGCCACG


 


266
UUGAGGUCUCAGGCAGCCA


 


267
UUUGAGGUCUCAGGCAGCC


 


268
UAUUGAGGUCUCAGGCAGC


 


269
UUAUUGAGGUCUCAGGCAG


 


270
UGUAUUGAGGUCUCAGGCA


 


271
UGGUAUUGAGGUCUCAGGC


 


272
UGGGUAUUGAGGUCUCAGG


 


273
UUAGGCAGGUGGACUUGGG


 


274
UAUAGGCAGGUGGACUUGG


 


275
UGAUAGGCAGGUGGACUUG


 


276
UGGAUAGGCAGGUGGACUU


 


277
UUGGAUAGGCAGGUGGACU


 


278
UAUGGAUAGGCAGGUGGAC


 


279
UGAUGGAUAGGCAGGUGGA


 


280
UGGAUGGAUAGGCAGGUGG


 


281
UAGGAUGGAUAGGCAGGUG


 


282
UCAGGAUGGAUAGGCAGGU


 


283
UGCAGGAUGGAUAGGCAGG


 


284
UCGCAGGAUGGAUAGGCAG


 


285
UUCGCAGGAUGGAUAGGCA


 


286
UCUCGCAGGAUGGAUAGGC


 


287
UGCUCGCAGGAUGGAUAGG


 


288
UAGCUCGCAGGAUGGAUAG


 


289
UGAGCUCGCAGGAUGGAUA


 


290
UGGAGCUCGCAGGAUGGAU


 


291
UAGGAGCUCGCAGGAUGGA


 


292
UAAGGAGCUCGCAGGAUGG


 


293
UCAAGGAGCUCGCAGGAUG


 


294
UCCAAGGAGCUCGCAGGAU


 


295
UCCCAAGGAGCUCGCAGGA


 


296
UACCCAAGGAGCUCGCAGG


 


297
UGACCCAAGGAGCUCGCAG


 


298
UGGACCCAAGGAGCUCGCA


 


299
UAGGACCCAAGGAGCUCGC


 


300
UCAGGACCCAAGGAGCUCG


 


301
UGCAGGACCCAAGGAGCUC


 


302
UUGCAGGACCCAAGGAGCU


 


303
UUUGCAGGACCCAAGGAGC


 


304
UAUUGCAGGACCCAAGGAG


 


305
UGAUUGCAGGACCCAAGGA


 


306
UAGAUUGCAGGACCCAAGG


 


307
UGAGAUUGCAGGACCCAAG


 


308
UGGAGAUUGCAGGACCCAA


 


309
UUGGAGAUUGCAGGACCCA


 


310
UCUGGAGAUUGCAGGACCC


 


311
UCCUGGAGAUUGCAGGACC


 


312
UCCCUGGAGAUUGCAGGAC


 


313
UGCCCUGGAGAUUGCAGGA


 


314
UAGCCCUGGAGAUUGCAGG


 


315
UCAGCCCUGGAGAUUGCAG


 


316
UGCAGCCCUGGAGAUUGCA


 


317
UGGCAGCCCUGGAGAUUGC


 


318
UGGGCAGCCCUGGAGAUUG


 


319
UUUUAAGCAACCUACAGGG


 


320
UUUUUAAGCAACCUACAGG


 


321
UCUUUUAAGCAACCUACAG


 


322
UCCUUUUAAGCAACCUACA


 


323
UCCCUUUUAAGCAACCUAC


 


324
UUCCCUUUUAAGCAACCUA


 


325
UGUCCCUUUUAAGCAACCU


 


326
UACUGUCCCUUUUAAGCAA


 


327
UUACUGUCCCUUUUAAGCA


 


328
UAUACUGUCCCUUUUAAGC


 


329
UAAUACUGUCCCUUUUAAG


 


330
UGAAUACUGUCCCUUUUAA


 


331
UAGAAUACUGUCCCUUUUA


 


332
UGAGAAUACUGUCCCUUUU


 


333
UUGAGAAUACUGUCCCUUU


 


334
UCUGAGAAUACUGUCCCUU


 


335
UACUGAGAAUACUGUCCCU


 


336
UCACUGAGAAUACUGUCCC


 


337
UGCACUGAGAAUACUGUCC


 


338
UAGCACUGAGAAUACUGUC


 


339
UGAGCACUGAGAAUACUGU


 


340
UAGAGCACUGAGAAUACUG


 


341
UGAGAGCACUGAGAAUACU


 


342
UGGAGAGCACUGAGAAUAC


 


343
UAGGAGAGCACUGAGAAUA


 


344
UUAGGAGAGCACUGAGAAU


 


345
UGUAGGAGAGCACUGAGAA


 


346
UGGUAGGAGAGCACUGAGA


 


347
UGGGUAGGAGAGCACUGAG


 


348
UGGCCAGGCAUGAGGUGGG


 


349
UGGGCCAGGCAUGAGGUGG


 


350
UGCCAGCAUGCCUGGAGGG


 


351
UGGCCAGCAUGCCUGGAGG


 


352
UAGGCCAGCAUGCCUGGAG


 


353
UGAGGCCAGCAUGCCUGGA


 


354
UGGAGGCCAGCAUGCCUGG


 


355
UGGGAGGCCAGCAUGCCUG


 


356
UUGGGAGGCCAGCAUGCCU


 


357
UAUUGGGAGGCCAGCAUGC


 


358
UUAUUGGGAGGCCAGCAUG


 


359
UUUAUUGGGAGGCCAGCAU


 


360
UUUUAUUGGGAGGCCAGCA


 


361
UCUUUAUUGGGAGGCCAGC


 


362
UGCUUUAUUGGGAGGCCAG


 


363
UAGCUUUAUUGGGAGGCCA


 


364
UCAGCUUUAUUGGGAGGCC


 


365
UCCAGCUUUAUUGGGAGGC


 


366
UUCCAGCUUUAUUGGGAGG


 


367
UGUCCAGCUUUAUUGGGAG


 


368
UUUGUCCAGCUUUAUUGGG


 


369
UCUUGUCCAGCUUUAUUGG


 


370
UUCUUGUCCAGCUUUAUUG


 


371
UUUCUUGUCCAGCUUUAUU


 


372
UCUUCUUGUCCAGCUUUAU


 


373
UGCUUCUUGUCCAGCUUUA


 


374
UGCAGCUUCUUGUCCAGCU


 


375
UUAGCAGCUUCUUGUCCAG


 


376
UAUAGCAGCUUCUUGUCCA 
















TABLE 1b







Nucleobase sequences and sugar-phosphate backbone 


modifications of the antisense strands of 376 exemplary constructs:










SEQ




ID



#
NO:
Oligo Sequence (5′ to 3′) and backbone modifications





 1
 804
PmU•fU•mC•fU•mA•fG•mG•fG•mA•fU•mG•fA•mA•fC•mU•fG•mA•fG•mC


 2
 805
PmU•fC•mU•fC•mU•fA•mG•fG•mG•fA•mU•fG•mA•fA•mC•fU•mG•fA•mG


 3
 806
PmU•fC•mC•fU•mC•fU•mA•fG•mG•fG•mA•fU•mG•fA•mA•fC•mU•fG•mA


 4
 807
PmU•fG•mC•fC•mU•fC•mU•fA•mG•fG•mG•fA•mU•fG•mA•fA•mC•fU•mG


 5
 808
PmU•fU•mG•fC•mC•fU•mC•fU•mA•fG•mG•fG•mA•fU•mG•fA•mA•fC•mU


 6
 809
PmU•fC•mU•fG•mC•fC•mU•fC•mU•fA•mG•fG•mG•fA•mU•fG•mA•fA•mC


 7
 810
PmU•fG•mC•fU•mG•fC•mC•fU•mC•fU•mA•fG•mG•fG•mA•fU•mG•fA•mA


 8
 811
PmU•fA•mG•fC•mU•fG•mC•fC•mU•fC•mU•fA•mG•fG•mG•fA•mU•fG•mA


 9
 812
PmU•fG•mC•fA•mG•fC•mU•fG•mC•fC•mU•fC•mU•fA•mG•fG•mG•fA•mU


 10
 813
PmU•fA•mG•fC•mA•fG•mC•fU•mG•fC•mC•fU•mC•fU•mA•fG•mG•fG•mA


 11
 814
PmU•fG•mA•fG•mC•fA•mG•fC•mU•fG•mC•fC•mU•fC•mU•fA•mG•fG•mG


 12
 815
PmU•fG•mG•fA•mG•fC•mA•fG•mC•fU•mG•fC•mC•fU•mC•fU•mA•fG•mG


 13
 816
PmU•fU•mG•fU•mU•fC•mC•fU•mG•fG•mA•fG•mC•fA•mG•fC•mU•fG•mC


 14
 817
PmU•fC•mU•fG•mU•fU•mC•fC•mU•fG•mG•fA•mG•fC•mA•fG•mC•fU•mG


 15
 818
PmU•fC•mC•fU•mC•fU•mG•fU•mU•fC•mC•fU•mG•fG•mA•fG•mC•fA•mG


 16
 819
PmU•fA•mC•fC•mU•fC•mU•fG•mU•fU•mC•fC•mU•fG•mG•fA•mG•fC•mA


 17
 820
PmU•fC•mA•fC•mC•fU•mC•fU•mG•fU•mU•fC•mC•fU•mG•fG•mA•fG•mC


 18
 821
PmU•fG•mC•fA•mC•fC•mU•fC•mU•fG•mU•fU•mC•fC•mU•fG•mG•fA•mG


 19
 822
PmU•fG•mG•fC•mA•fC•mC•fU•mC•fU•mG•fU•mU•fC•mC•fU•mG•fG•mA


 20
 823
PmU•fU•mG•fG•mC•fA•mC•fC•mU•fC•mU•fG•mU•fU•mC•fC•mU•fG•mG


 21
 824
PmU•fA•mU•fG•mG•fC•mA•fC•mC•fU•mC•fU•mG•fU•mU•fC•mC•fU•mG


 22
 825
PmU•fC•mA•fU•mG•fG•mC•fA•mC•fC•mU•fC•mU•fG•mU•fU•mC•fC•mU


 23
 826
PmU•fU•mG•fC•mA•fU•mG•fG•mC•fA•mC•fC•mU•fC•mU•fG•mU•fU•mC


 24
 827
PmU•fC•mU•fG•mC•fA•mU•fG•mG•fC•mA•fC•mC•fU•mC•fU•mG•fU•mU


 25
 828
PmU•fG•mC•fU•mG•fC•mA•fU•mG•fG•mC•fA•mC•fC•mU•fC•mU•fG•mU


 26
 829
PmU•fG•mG•fC•mU•fG•mC•fA•mU•fG•mG•fC•mA•fC•mC•fU•mC•fU•mG


 27
 830
PmU•fG•mG•fG•mC•fU•mG•fC•mA•fU•mG•fG•mC•fA•mC•fC•mU•fC•mU


 28
 831
PmU•fC•mA•fA•mC•fA•mA•fG•mG•fA•mG•fU•mA•fC•mC•fC•mG•fG•mG


 29
 832
PmU•fA•mC•fA•mA•fC•mA•fA•mG•fG•mA•fG•mU•fA•mC•fC•mC•fG•mG


 30
 833
PmU•fA•mA•fC•mA•fA•mC•fA•mA•fG•mG•fA•mG•fU•mA•fC•mC•fC•mG


 31
 834
PmU•fC•mA•fA•mC•fA•mA•fC•mA•fA•mG•fG•mA•fG•mU•fA•mC•fC•mC


 32
 835
PmU•fG•mC•fA•mA•fC•mA•fA•mC•fA•mA•fG•mG•fA•mG•fU•mA•fC•mC


 33
 836
PmU•fG•mG•fC•mA•fA•mC•fA•mA•fC•mA•fA•mG•fG•mA•fG•mU•fA•mC


 34
 837
PmU•fG•mG•fG•mC•fA•mA•fC•mA•fA•mC•fA•mA•fG•mG•fA•mG•fU•mA


 35
 838
PmU•fA•mG•fG•mG•fC•mA•fA•mC•fA•mA•fC•mA•fA•mG•fG•mA•fG•mU


 36
 839
PmU•fG•mA•fG•mG•fG•mC•fA•mA•fC•mA•fA•mC•fA•mA•fG•mG•fA•mG


 37
 840
PmU•fG•mG•fA•mG•fG•mG•fC•mA•fA•mC•fA•mA•fC•mA•fA•mG•fG•mA


 38
 841
PmU•fA•mG•fG•mA•fG•mG•fG•mC•fA•mA•fC•mA•fA•mC•fA•mA•fG•mG


 39
 842
PmU•fC•mA•fG•mG•fA•mG•fG•mG•fC•mA•fA•mC•fA•mA•fC•mA•fA•mG


 40
 843
PmU•fC•mC•fA•mG•fG•mA•fG•mG•fG•mC•fA•mA•fC•mA•fA•mC•fA•mA


 41
 844
PmU•fG•mC•fC•mA•fG•mG•fA•mG•fG•mG•fC•mA•fA•mC•fA•mA•fC•mA


 42
 845
PmU•fC•mG•fC•mC•fA•mG•fG•mA•fG•mG•fG•mC•fA•mA•fC•mA•fA•mC


 43
 846
PmU•fG•mC•fG•mC•fC•mA•fG•mG•fA•mG•fG•mG•fC•mA•fA•mC•fA•mA


 44
 847
PmU•fA•mG•fC•mG•fC•mC•fA•mG•fG•mA•fG•mG•fG•mC•fA•mA•fC•mA


 45
 848
PmU•fG•mA•fG•mC•fG•mC•fC•mA•fG•mG•fA•mG•fG•mG•fC•mA•fA•mC


 46
 849
PmU•fG•mG•fA•mG•fC•mG•fC•mC•fA•mG•fG•mA•fG•mG•fG•mC•fA•mA


 47
 850
PmU•fA•mG•fG•mA•fG•mC•fG•mC•fC•mA•fG•mG•fA•mG•fG•mG•fC•mA


 48
 851
PmU•fG•mC•fC•mA•fG•mG•fA•mG•fC•mG•fC•mC•fA•mG•fG•mA•fG•mG


 49
 852
PmU•fA•mG•fA•mG•fG•mC•fC•mA•fG•mG•fA•mG•fC•mG•fC•mC•fA•mG


 50
 853
PmU•fC•mA•fG•mA•fG•mG•fC•mC•fA•mG•fG•mA•fG•mC•fG•mC•fC•mA


 51
 854
PmU•fG•mC•fA•mG•fA•mG•fG•mC•fC•mA•fG•mG•fA•mG•fC•mG•fC•mC


 52
 855
PmU•fG•mG•fC•mA•fG•mA•fG•mG•fC•mC•fA•mG•fG•mA•fG•mC•fG•mC


 53
 856
PmU•fG•mG•fG•mC•fA•mG•fA•mG•fG•mC•fC•mA•fG•mG•fA•mG•fC•mG


 54
 857
PmU•fU•mC•fG•mG•fG•mC•fA•mG•fA•mG•fG•mC•fC•mA•fG•mG•fA•mG


 55
 858
PmU•fC•mU•fC•mG•fG•mG•fC•mA•fG•mA•fG•mG•fC•mC•fA•mG•fG•mA


 56
 859
PmU•fG•mC•fU•mC•fG•mG•fG•mC•fA•mG•fA•mG•fG•mC•fC•mA•fG•mG


 57
 860
PmU•fA•mG•fC•mU•fC•mG•fG•mG•fC•mA•fG•mA•fG•mG•fC•mC•fA•mG


 58
 861
PmU•fA•mA•fG•mC•fU•mC•fG•mG•fG•mC•fA•mG•fA•mG•fG•mC•fC•mA


 59
 862
PmU•fG•mA•fA•mG•fC•mU•fC•mG•fG•mG•fC•mA•fG•mA•fG•mG•fC•mC


 60
 863
PmU•fU•mG•fA•mA•fG•mC•fU•mC•fG•mG•fG•mC•fA•mG•fA•mG•fG•mC


 61
 864
PmU•fC•mU•fG•mA•fA•mG•fC•mU•fC•mG•fG•mG•fC•mA•fG•mA•fG•mG


 62
 865
PmU•fU•mC•fU•mG•fA•mA•fG•mC•fU•mC•fG•mG•fG•mC•fA•mG•fA•mG


 63
 866
PmU•fC•mU•fC•mU•fG•mA•fA•mG•fC•mU•fC•mG•fG•mG•fC•mA•fG•mA


 64
 867
PmU•fC•mC•fU•mC•fU•mG•fA•mA•fG•mC•fU•mC•fG•mG•fG•mC•fA•mG


 65
 868
PmU•fG•mC•fC•mU•fC•mU•fG•mA•fA•mG•fC•mU•fC•mG•fG•mG•fC•mA


 66
 869
PmU•fG•mG•fC•mC•fU•mC•fU•mG•fA•mA•fG•mC•fU•mC•fG•mG•fG•mC


 67
 870
PmU•fC•mG•fG•mC•fC•mU•fC•mU•fG•mA•fA•mG•fC•mU•fC•mG•fG•mG


 68
 871
PmU•fU•mC•fG•mG•fC•mC•fU•mC•fU•mG•fA•mA•fG•mC•fU•mC•fG•mG


 69
 872
PmU•fC•mU•fC•mG•fG•mC•fC•mU•fC•mU•fG•mA•fA•mG•fC•mU•fC•mG


 70
 873
PmU•fC•mC•fU•mC•fG•mG•fC•mC•fU•mC•fU•mG•fA•mA•fG•mC•fU•mC


 71
 874
PmU•fU•mC•fC•mU•fC•mG•fG•mC•fC•mU•fC•mU•fG•mA•fA•mG•fC•mU


 72
 875
PmU•fA•mU•fC•mC•fU•mC•fG•mG•fC•mC•fU•mC•fU•mG•fA•mA•fG•mC


 73
 876
PmU•fC•mA•fU•mC•fC•mU•fC•mG•fG•mC•fC•mU•fC•mU•fG•mA•fA•mG


 74
 877
PmU•fG•mC•fA•mU•fC•mC•fU•mC•fG•mG•fC•mC•fU•mC•fU•mG•fA•mA


 75
 878
PmU•fG•mG•fC•mA•fU•mC•fC•mU•fC•mG•fG•mC•fC•mU•fC•mU•fG•mA


 76
 879
PmU•fA•mG•fG•mC•fA•mU•fC•mC•fU•mC•fG•mG•fC•mC•fU•mC•fU•mG


 77
 880
PmU•fG•mA•fG•mG•fC•mA•fU•mC•fC•mU•fC•mG•fG•mC•fC•mU•fC•mU


 78
 881
PmU•fG•mG•fA•mG•fG•mC•fA•mU•fC•mC•fU•mC•fG•mG•fC•mC•fU•mC


 79
 882
PmU•fG•mG•fG•mA•fG•mG•fC•mA•fU•mC•fC•mU•fC•mG•fG•mC•fC•mU


 80
 883
PmU•fA•mG•fG•mG•fA•mG•fG•mC•fA•mU•fC•mC•fU•mC•fG•mG•fC•mC


 81
 884
PmU•fA•mA•fG•mG•fG•mA•fG•mG•fC•mA•fU•mC•fC•mU•fC•mG•fG•mC


 82
 885
PmU•fG•mA•fA•mG•fG•mG•fA•mG•fG•mC•fA•mU•fC•mC•fU•mC•fG•mG


 83
 886
PmU•fA•mG•fA•mA•fG•mG•fG•mA•fG•mG•fC•mA•fU•mC•fC•mU•fC•mG


 84
 887
PmU•fG•mA•fG•mA•fA•mG•fG•mG•fA•mG•fG•mC•fA•mU•fC•mC•fU•mC


 85
 888
PmU•fU•mG•fA•mG•fA•mA•fG•mG•fG•mA•fG•mG•fC•mA•fU•mC•fC•mU


 86
 889
PmU•fC•mU•fG•mA•fG•mA•fA•mG•fG•mG•fA•mG•fG•mC•fA•mU•fC•mC


 87
 890
PmU•fG•mC•fU•mG•fA•mG•fA•mA•fG•mG•fG•mA•fG•mG•fC•mA•fU•mC


 88
 891
PmU•fA•mG•fC•mU•fG•mA•fG•mA•fA•mG•fG•mG•fA•mG•fG•mC•fA•mU


 89
 892
PmU•fU•mG•fA•mA•fG•mC•fU•mG•fA•mG•fA•mA•fG•mG•fG•mA•fG•mG


 90
 893
PmU•fA•mU•fG•mA•fA•mG•fC•mU•fG•mA•fG•mA•fA•mG•fG•mG•fA•mG


 91
 894
PmU•fC•mA•fU•mG•fA•mA•fG•mC•fU•mG•fA•mG•fA•mA•fG•mG•fG•mA


 92
 895
PmU•fG•mC•fA•mU•fG•mA•fA•mG•fC•mU•fG•mA•fG•mA•fA•mG•fG•mG


 93
 896
PmU•fU•mG•fC•mA•fU•mG•fA•mA•fG•mC•fU•mG•fA•mG•fA•mA•fG•mG


 94
 897
PmU•fC•mU•fG•mC•fA•mU•fG•mA•fA•mG•fC•mU•fG•mA•fG•mA•fA•mG


 95
 898
PmU•fC•mC•fU•mG•fC•mA•fU•mG•fA•mA•fG•mC•fU•mG•fA•mG•fA•mA


 96
 899
PmU•fC•mC•fC•mU•fG•mC•fA•mU•fG•mA•fA•mG•fC•mU•fG•mA•fG•mA


 97
 900
PmU•fA•mC•fC•mC•fU•mG•fC•mA•fU•mG•fA•mA•fG•mC•fU•mG•fA•mG


 98
 901
PmU•fA•mA•fC•mC•fC•mU•fG•mC•fA•mU•fG•mA•fA•mG•fC•mU•fG•mA


 99
 902
PmU•fU•mA•fA•mC•fC•mC•fU•mG•fC•mA•fU•mG•fA•mA•fG•mC•fU•mG


100
 903
PmU•fG•mU•fA•mA•fC•mC•fC•mU•fG•mC•fA•mU•fG•mA•fA•mG•fC•mU


101
 904
PmU•fU•mG•fU•mA•fA•mC•fC•mC•fU•mG•fC•mA•fU•mG•fA•mA•fG•mC


102
 905
PmU•fA•mU•fG•mU•fA•mA•fC•mC•fC•mU•fG•mC•fA•mU•fG•mA•fA•mG


103
 906
PmU•fC•mA•fU•mG•fU•mA•fA•mC•fC•mC•fU•mG•fC•mA•fU•mG•fA•mA


104
 907
PmU•fU•mC•fA•mU•fG•mU•fA•mA•fC•mC•fC•mU•fG•mC•fA•mU•fG•mA


105
 908
PmU•fU•mU•fC•mA•fU•mG•fU•mA•fA•mC•fC•mC•fU•mG•fC•mA•fU•mG


106
 909
PmU•fC•mU•fU•mC•fA•mU•fG•mU•fA•mA•fC•mC•fC•mU•G•mC•fA•mU


107
 910
PmU•fG•mC•fU•mU•fC•mA•fU•mG•fU•mA•fA•mC•fC•mC•fU•mG•fC•mA


108
 911
PmU•fU•mG•fC•mU•fU•mC•fA•mU•fG•mU•fA•mA•fC•mC•fC•mU•fG•mC


109
 912
PmU•fG•mU•fG•mC•fU•mU•fC•mA•fU•mG•fU•mA•fA•mC•fC•mC•fU•mG


110
 913
PmU•fC•mG•fU•mG•fC•mU•fU•mC•fA•mU•fG•mU•fA•mA•fC•mC•fC•mU


111
 914
PmU•fG•mC•fG•mU•fG•mC•fU•mU•fC•mA•fU•mG•fU•mA•fA•mC•fC•mC


112
 915
PmU•fG•mG•fC•mG•fU•mG•fC•mU•fU•mC•fA•mU•fG•mU•fA•mA•fC•mC


113
 916
PmU•fU•mG•fG•mC•fG•mU•fG•mC•fU•mU•fC•mA•fU•mG•fU•mA•fA•mC


114
 917
PmU•fG•mU•fG•mG•fC•mG•fU•mG•fC•mU•fU•mC•fA•mU•fG•mU•fA•mA


115
 918
PmU•fG•mG•fU•mG•fG•mC•fG•mU•fG•mC•fU•mU•fC•mA•fU•mG•fU•mA


116
 919
PmU•fU•mG•fG•mU•fG•mG•fC•mG•fU•mG•fC•mU•fU•mC•fA•mU•fG•mU


117
 920
PmU•fU•mU•fG•mG•fU•mG•fG•mC•fG•mU•fG•mC•fU•mU•fC•mA•fU•mG


118
 921
PmU•fC•mU•fU•mG•fG•mU•fG•mG•fC•mG•fU•mG•fC•mU•fU•mC•fA•mU


119
 922
PmU•fU•mC•fU•mU•fG•mG•fU•mG•fG•mC•fG•mU•fG•mC•fU•mU•fC•mA


120
 923
PmU•fG•mU•fC•mU•fU•mG•fG•mU•fG•mG•fC•mG•fU•mG•fC•mU•fU•mC


121
 924
PmU•fG•mG•fU•mC•fU•mU•fG•mG•fU•mG•fG•mC•fG•mU•fG•mC•fU•mU


122
 925
PmU•fC•mG•fG•mU•fC•mU•fU•mG•fG•mU•fG•mG•fC•mG•fU•mG•fC•mU


123
 926
PmU•fG•mC•fG•mG•fU•mC•fU•mU•fG•mG•fU•mG•fG•mC•fG•mU•fG•mC


124
 927
PmU•fG•mG•fC•mG•fG•mU•fC•mU•fU•mG•fG•mU•fG•mG•fC•mG•fU•mG


125
 928
PmU•fU•mG•fG•mC•fG•mG•fU•mC•fU•mU•fG•mG•fU•mG•fG•mC•fG•mU


126
 929
PmU•fU•mU•fG•mG•fC•mG•fG•mU•fC•mU•fU•mG•fG•mU•fG•mG•fC•mG


127
 930
PmU•fC•mU•fU•mG•fG•mC•fG•mG•fU•mC•fU•mU•fG•mG•fU•mG•fG•mC


128
 931
PmU•fC•mC•fU•mU•fG•mG•fC•mG•fG•mU•fC•mU•fU•mG•fG•mU•fG•mG


129
 932
PmU•fU•mC•fC•mU•fU•mG•fG•mC•fG•mG•fU•mC•fU•mU•fG•mG•fU•mG


130
 933
PmU•fA•mU•fC•mC•fU•mU•fG•mG•fC•mG•fG•mU•fC•mU•fU•mG•fG•mU


131
 934
PmU•fC•mA•fU•mC•fC•mU•fU•mG•fG•mC•fG•mG•fU•mC•fU•mU•fG•mG


132
 935
PmU•fG•mC•fA•mU•fC•mC•fU•mU•fG•mG•fC•mG•fG•mU•fC•mU•fU•mG


133
 936
PmU•fU•mG•fC•mA•fU•mC•fC•mU•fU•mG•fG•mC•fG•mG•fU•mC•fU•mU


134
 937
PmU•fG•mU•fG•mC•fA•mU•fC•mC•fU•mU•fG•mG•fC•mG•fG•mU•fC•mU


135
 938
PmU•fA•mG•fU•mG•fC•mA•fU•mC•fC•mU•fU•mG•fG•mC•fG•mG•fU•mC


136
 939
PmU•fC•mA•fG•mU•fG•mC•fA•mU•fC•mC•fU•mU•fG•mG•fC•mG•fG•mU


137
 940
PmU•fU•mC•fA•mG•fU•mG•fC•mA•fU•mC•fC•mU•fU•mG•fG•mC•fG•mG


138
 941
PmU•fC•mU•fC•mA•fG•mU•fG•mC•fA•mU•fC•mC•fU•mU•fG•mG•fC•mG


139
 942
PmU•fG•mC•fU•mC•fA•mG•fU•mG•fC•mA•fU•mC•fC•mU•fU•mG•fG•mC


140
 943
PmU•fU•mG•fC•mU•fC•mA•fG•mU•fG•mC•fA•mU•fC•mC•fU•mU•fG•mG


141
 944
PmU•fC•mU•fG•mC•fU•mC•fA•mG•fU•mG•fC•mA•fU•mC•fC•mU•fU•mG


142
 945
PmU•fG•mC•fU•mG•fC•mU•fC•mA•fG•mU•fG•mC•fA•mU•fC•mC•fU•mU


143
 946
PmU•fC•mG•fC•mU•fG•mC•fU•mC•fA•mG•fU•mG•fC•mA•fU•mC•fC•mU


144
 947
PmU•fA•mC•fG•mC•fU•mG•fC•mU•fC•mA•fG•mU•fG•mC•fA•mU•fC•mC


145
 948
PmU•fC•mA•fC•mG•fC•mU•fG•mC•fU•mC•fA•mG•fU•mG•fC•mA•fU•mC


146
 949
PmU•fG•mC•fA•mC•fG•mC•fU•mG•fC•mU•fC•mA•fG•mU•fG•mC•fA•mU


147
 950
PmU•fU•mG•fC•mA•fC•mG•fC•mU•fG•mC•fU•mC•fA•mG•fU•mG•fC•mA


148
 951
PmU•fC•mU•fG•mC•fA•mC•fG•mC•fU•mG•fC•mU•fC•mA•fG•mU•fG•mC


149
 952
PmU•fC•mC•fU•mG•fC•mA•fC•mG•fC•mU•fG•mC•fU•mC•fA•mG•fU•mG


150
 953
PmU•fU•mC•fC•mU•fG•mC•fA•mC•fG•mC•fU•mG•fC•mU•fC•mA•fG•mU


151
 954
PmU•fA•mC•fU•mC•fC•mU•fG•mC•fA•mC•fG•mC•fU•mG•fC•mU•fC•mA


152
 955
PmU•fG•mG•fG•mA•fC•mU•fC•mC•fU•mG•fC•mA•fC•mG•fC•mU•fG•mC


153
 956
PmU•fU•mG•fG•mG•fA•mC•fU•mC•fC•mU•fG•mC•fA•mC•fG•mC•fU•mG


154
 957
PmU•fC•mU•fG•mG•fG•mA•fC•mU•fC•mC•fU•mG•fC•mA•fC•mG•fC•mU


155
 958
PmU•fC•mC•fU•mG•fG•mG•fA•mC•fU•mC•fC•mU•fG•mC•fA•mC•fG•mC


156
 959
PmU•fA•mC•fC•mU•fG•mG•fG•mA•fC•mU•fC•mC•fU•mG•fC•mA•fC•mG


157
 960
PmU•fC•mA•fC•mC•fU•mG•fG•mG•fA•mC•fU•mC•fC•mU•fG•mC•fA•mC


158
 961
PmU•fC•mC•fA•mC•fC•mU•fG•mG•fG•mA•fC•mU•fC•mC•fU•mG•fC•mA


159
 962
PmU•fG•mG•fG•mC•fC•mA•fC•mC•fU•mG•fG•mG•fA•mC•fU•mC•fC•mU


160
 963
PmU•fU•mG•fG•mG•fC•mC•fA•mC•fC•mU•fG•mG•fG•mA•fC•mU•fC•mC


161
 964
PmU•fU•mG•fC•mU•fG•mG•fG•mC•fC•mA•fC•mC•fU•mG•fG•mG•fA•mC


162
 965
PmU•fC•mU•fG•mC•fU•mG•fG•mG•fC•mC•fA•mC•fC•mU•fG•mG•fG•mA


163
 966
PmU•fG•mG•fC•mC•fU•mG•fC•mU•fG•mG•fG•mC•fC•mA•fC•mC•fU•mG


164
 967
PmU•fC•mC•fU•mG•fG•mC•fC•mU•fG•mC•fU•mG•fG•mG•fC•mC•fA•mC


165
 968
PmU•fC•mC•fA•mU•fC•mG•fG•mU•fC•mA•fC•mC•fC•mA•fG•mC•fC•mC


166
 969
PmU•fG•mC•fC•mA•fU•mC•fG•mG•fU•mC•fA•mC•fC•mC•fA•mG•fC•mC


167
 970
PmU•fA•mG•fC•mC•fA•mU•fC•mG•fG•mU•fC•mA•fC•mC•fC•mA•fG•mC


168
 971
PmU•fA•mA•fG•mC•fC•mA•fU•mC•fG•mG•fU•mC•fA•mC•fC•mC•fA•mG


169
 972
PmU•fG•mA•fA•mG•fC•mC•fA•mU•fC•mG•fG•mU•fC•mA•fC•mC•fC•mA


170
 973
PmU•fU•mG•fA•mA•fG•mC•fC•mA•fU•mC•fG•mG•fU•mC•fA•mC•fC•mC


171
 974
PmU•fC•mU•fG•mA•fA•mG•fC•mC•fA•mU•fC•mG•fG•mU•fC•mA•fC•mC


172
 975
PmU•fA•mC•fU•mG•fA•mA•fG•mC•fC•mA•fU•mC•fG•mG•fU•mC•fA•mC


173
 976
PmU•fA•mA•fC•mU•fG•mA•fA•mG•fC•mC•fA•mU•fC•mG•fG•mU•fC•mA


174
 977
PmU•fG•mA•fA•mC•fU•mG•fA•mA•fG•mC•fC•mA•fU•mC•fG•mG•fU•mC


175
 978
PmU•fG•mG•fA•mA•fC•mU•fG•mA•fA•mG•fC•mC•fA•mU•fC•mG•fG•mU


176
 979
PmU•fG•mG•fG•mA•fA•mC•fU•mG•fA•mA•fG•mC•fC•mA•fU•mC•fG•mG


177
 980
PmU•fA•mG•fG•mG•fA•mA•fC•mU•fG•mA•fA•mG•fC•mC•fA•mU•fC•mG


178
 981
PmU•fC•mA•fG•mG•fG•mA•fA•mC•fU•mG•fA•mA•fG•mC•fC•mA•fU•mC


179
 982
PmU•fU•mC•fA•mG•fG•mG•fA•mA•fC•mU•fG•mA•fA•mG•fC•mC•fA•mU


180
 983
PmU•fU•mU•fC•mA•fG•mG•fG•mA•fA•mC•fU•mG•fA•mA•fG•mC•fC•mA


181
 984
PmU•fU•mU•fU•mC•fA•mG•fG•mG•fA•mA•fC•mU•fG•mA•fA•mG•fC•mC


182
 985
PmU•fC•mU•fU•mU•fC•mA•fG•mG•fG•mA•fA•mC•fU•mG•fA•mA•fG•mC


183
 986
PmU•fU•mC•fU•mU•fU•mC•fA•mG•fG•mG•fA•mA•fC•mU•fG•mA•fA•mG


184
 987
PmU•fG•mU•fC•mU•fU•mU•fC•mA•fG•mG•fG•mA•fA•mC•fU•mG•fA•mA


185
 988
PmU•fA•mG•fU•mC•fU•mU•fU•mC•fA•mG•fG•mG•fA•mA•fC•mU•fG•mA


186
 989
PmU•fU•mA•fG•mU•fC•mU•fU•mU•fC•mA•fG•mG•fG•mA•fA•mC•fU•mG


187
 990
PmU•fG•mU•fA•mG•fU•mC•fU•mU•fU•mC•fA•mG•fG•mG•fA•mA•fC•mU


188
 991
PmU•fA•mG•fU•mA•fG•mU•fC•mU•fU•mU•fC•mA•fG•mG•fG•mA•fA•mC


189
 992
PmU•fC•mA•fG•mU•fA•mG•fU•mC•fU•mU•fU•mC•fA•mG•fG•mG•fA•mA


190
 993
PmU•fC•mC•fA•mG•fU•mA•fG•mU•fC•mU•fU•mU•fC•mA•fG•mG•fG•mA


191
 994
PmU•fU•mC•fC•mA•fG•mU•fA•mG•fU•mC•fU•mU•fU•mC•fA•mG•fG•mG


192
 995
PmU•fC•mU•fC•mC•fA•mG•fU•mA•fG•mU•fC•mU•fU•mU•fC•mA•fG•mG


193
 996
PmU•fG•mC•fU•mC•fC•mA•fG•mU•fA•mG•fU•mC•fU•mU•fU•mC•fA•mG


194
 997
PmU•fU•mG•fC•mU•fC•mC•fA•mG•fU•mA•fG•mU•fC•mU•fU•mU•fC•mA


195
 998
PmU•fG•mU•fG•mC•fU•mC•fC•mA•fG•mU•fA•mG•fU•mC•fU•mU•fU•mC


196
 999
PmU•fG•mG•fU•mG•fC•mU•fC•mC•fA•mG•fU•mA•fG•mU•fC•mU•fU•mU


197
1000
PmU•fC•mG•fG•mU•fG•mC•fU•mC•fC•mA•fG•mU•fA•mG•fU•mC•fU•mU


198
1001
PmU•fA•mC•fG•mG•fU•mG•fC•mU•fC•mC•fA•mG•fU•mA•fG•mU•fC•mU


199
1002
PmU•fA•mA•fC•mG•fG•mU•fG•mC•fU•mC•fC•mA•fG•mU•fA•mG•fU•mC


200
1003
PmU•fU•mA•fA•mC•fG•mG•fU•mG•fC•mU•fC•mC•fA•mG•fU•mA•fG•mU


201
1004
PmU•fU•mU•fA•mA•fC•mG•fG•mU•fG•mC•fU•mC•fC•mA•fG•mU•fA•mG


202
1005
PmU•fC•mU•fU•mA•fA•mC•fG•mG•fU•mG•fC•mU•fC•mC•fA•mG•fU•mA


203
1006
PmU•fC•mC•fU•mU•fA•mA•fC•mG•fG•mU•fG•mC•fU•mC•fC•mA•fG•mU


204
1007
PmU•fU•mC•fC•mU•fU•mA•fA•mC•fG•mG•fU•mG•fC•mU•fC•mC•fA•mG


205
1008
PmU•fG•mU•fC•mC•fU•mU•fA•mA•fC•mG•fG•mU•fG•mC•fU•mC•fC•mA


206
1009
PmU•fU•mG•fU•mC•fC•mU•fU•mA•fA•mC•fG•mG•fU•mG•fC•mU•fC•mC


207
1010
PmU•fU•mU•fG•mU•fC•mC•fU•mU•fA•mA•fC•mG•fG•mU•fG•mC•fU•mC


208
1011
PmU•fC•mU•fU•mG•fU•mC•fC•mU•fU•mA•fA•mC•fG•mG•fU•mG•fC•mU


209
1012
PmU•fA•mC•fU•mU•fG•mU•fC•mC•fU•mU•fA•mA•fC•mG•fG•mU•fG•mC


210
1013
PmU•fA•mA•fC•mU•fU•mG•fU•mC•fC•mU•fU•mA•fA•mC•fG•mG•fU•mG


211
1014
PmU•fG•mA•fA•mC•fU•mU•fG•mU•fC•mC•fU•mU•fA•mA•fC•mG•fG•mU


212
1015
PmU•fA•mG•fA•mA•fC•mU•fU•mG•fU•mC•fC•mU•fU•mA•fA•mC•fG•mG


213
1016
PmU•fG•mA•fG•mA•fA•mC•fU•mU•fG•mU•fC•mC•fU•mU•fA•mA•fC•mG


214
1017
PmU•fA•mG•fA•mG•fA•mA•fC•mU•fU•mG•fU•mC•fC•mU•fU•mA•fA•mC


215
1018
PmU•fC•mA•fG•mA•fG•mA•fA•mC•fU•mU•fG•mU•fC•mC•fU•mU•fA•mA


216
1019
PmU•fU•mC•fA•mG•fA•mG•fA•mA•fC•mU•fU•mG•fU•mC•fC•mU•fU•mA


217
1020
PmU•fC•mU•fC•mA•fG•mA•fG•mA•fA•mC•fU•mU•fG•mU•fC•mC•fU•mU


218
1021
PmU•fA•mC•fU•mC•fA•mG•fA•mG•fA•mA•fC•mU•fU•mG•fU•mC•fC•mU


219
1022
PmU•fA•mA•fC•mU•fC•mA•fG•mA•fG•mA•fA•mC•fU•mU•fG•mU•fC•mC


220
1023
PmU•fG•mA•fA•mC•fU•mC•fA•mG•fA•mG•fA•mA•fC•mU•fU•mG•fU•mC


221
1024
PmU•fC•mA•fG•mA•fA•mC•fU•mC•fA•mG•fA•mG•fA•mA•fC•mU•fU•mG


222
1025
PmU•fC•mC•fA•mG•fA•mA•fC•mU•fC•mA•fG•mA•fG•mA•fA•mC•fU•mU


223
1026
PmU•fC•mC•fC•mA•fG•mA•fA•mC•fU•mC•fA•mG•fA•mG•fA•mA•fC•mU


224
1027
PmU•fU•mC•fC•mC•fA•mG•fA•mA•fC•mU•fC•mA•fG•mA•fG•mA•fA•mC


225
1028
PmU•fA•mU•fC•mC•fC•mA•fG•mA•fA•mC•fU•mC•fA•mG•fA•mG•fA•mA


226
1029
PmU•fA•mA•fU•mC•fC•mC•fA•mG•fA•mA•fC•mU•fC•mA•fG•mA•fG•mA


227
1030
PmU•fA•mA•fA•mU•fC•mC•fC•mA•fG•mA•fA•mC•fU•mC•fA•mG•fA•mG


228
1031
PmU•fC•mA•fA•mA•fU•mC•fC•mC•fA•mG•fA•mA•fC•mU•fC•mA•fG•mA


229
1032
PmU•fC•mC•fA•mA•fA•mU•fC•mC•fC•mA•fG•mA•fA•mC•fU•mC•fA•mG


230
1033
PmU•fU•mC•fC•mA•fA•mA•fU•mC•fC•mC•fA•mG•fA•mA•fC•mU•fC•mA


231
1034
PmU•fG•mU•fC•mC•fA•mA•fA•mU•fC•mC•fC•mA•fG•mA•fA•mC•fU•mC


232
1035
PmU•fG•mG•fU•mC•fC•mA•fA•mA•fU•mC•fC•mC•fA•mG•fA•mA•fC•mU


233
1036
PmU•fG•mG•fG•mU•fC•mC•fA•mA•fA•mU•fC•mC•fC•mA•fG•mA•fA•mC


234
1037
PmU•fA•mG•fG•mG•fU•mC•fC•mA•fA•mA•fU•mC•fC•mC•fA•mG•fA•mA


235
1038
PmU•fC•mA•fG•mG•fG•mU•fC•mC•fA•mA•fA•mU•fC•mC•fC•mA•fG•mA


236
1039
PmU•fU•mC•fA•mG•fG•mG•fU•mC•fC•mA•fA•mA•fU•mC•fC•mC•fA•mG


237
1040
PmU•fG•mA•fC•mC•fU•mC•fA•mG•fG•mG•fU•mC•fC•mA•fA•mA•fU•mC


238
1041
PmU•fU•mG•fA•mC•fC•mU•fC•mA•fG•mG•fG•mU•fC•mC•fA•mA•fA•mU


239
1042
PmU•fC•mU•fG•mA•fC•mC•fU•mC•fA•mG•fG•mG•fU•mC•fC•mA•fA•mA


240
1043
PmU•fU•mC•fU•mG•fA•mC•fC•mU•fC•mA•fG•mG•fG•mU•fC•mC•fA•mA


241
1044
PmU•fG•mU•fC•mU•fG•mA•fC•mC•fU•mC•fA•mG•fG•mG•fU•mC•fC•mA


242
1045
PmU•fG•mG•fU•mC•fU•mG•fA•mC•fC•mU•fC•mA•fG•mG•fG•mU•fC•mC


243
1046
PmU•fU•mG•fG•mU•fC•mU•fG•mA•fC•mC•fU•mC•fA•mG•fG•mG•fU•mC


244
1047
PmU•fU•mU•fG•mG•fU•mC•fU•mG•fA•mC•fC•mU•fC•mA•fG•mG•fG•mU


245
1048
PmU•fG•mU•fU•mG•fG•mU•fC•mU•fG•mA•fC•mC•fU•mC•fA•mG•fG•mG


246
1049
PmU•fA•mG•fU•mU•fG•mG•fU•mC•fU•mG•fA•mC•fC•mU•fC•mA•fG•mG


247
1050
PmU•fA•mA•fG•mU•fU•mG•fG•mU•fC•mU•fG•mA•fC•mC•fU•mC•fA•mG


248
1051
PmU•fG•mA•fA•mG•fU•mU•fG•mG•fU•mC•fU•mG•fA•mC•fC•mU•fC•mA


249
1052
PmU•fU•mG•fA•mA•fG•mU•fU•mG•fG•mU•fC•mU•fG•mA•fC•mC•fU•mC


250
1053
PmU•fC•mU•fG•mA•fA•mG•fU•mU•fG•mG•fU•mC•fU•mG•fA•mC•fC•mU


251
1054
PmU•fG•mG•fC•mU•fG•mA•fA•mG•fU•mU•fG•mG•fU•mC•fU•mG•fA•mC


252
1055
PmU•fC•mG•fG•mC•fU•mG•fA•mA•fG•mU•fU•mG•fG•mU•fC•mU•fG•mA


253
1056
PmU•fA•mC•fG•mG•fC•mU•fG•mA•fA•mG•fU•mU•fG•mG•fU•mC•fU•mG


254
1057
PmU•fC•mA•fC•mG•fG•mC•fU•mG•fA•mA•fG•mU•fU•mG•fG•mU•fC•mU


255
1058
PmU•fC•mC•fA•mC•fG•mG•fC•mU•fG•mA•fA•mG•fU•mU•fG•mG•fU•mC


256
1059
PmU•fG•mC•fC•mA•fC•mG•fG•mC•fU•mG•fA•mA•fG•mU•fU•mG•fG•mU


257
1060
PmU•fC•mA•fG•mC•fC•mA•fC•mG•fG•mC•fU•mG•fA•mA•fG•mU•fU•mG


258
1061
PmU•fG•mC•fA•mG•fC•mC•fA•mC•fG•mG•fC•mU•fG•mA•fA•mG•fU•mU


259
1062
PmU•fG•mG•fC•mA•fG•mC•fC•mA•fC•mG•fG•mC•fU•mG•fA•mA•fG•mU


260
1063
PmU•fA•mG•fG•mC•fA•mG•fC•mC•fA•mC•fG•mG•fC•mU•fG•mA•fA•mG


261
1064
PmU•fC•mA•fG•mG•fC•mA•fG•mC•fC•mA•fC•mG•fG•mC•fU•mG•fA•mA


262
1065
PmU•fU•mC•fU•mC•fA•mG•fG•mC•fA•mG•fC•mC•fA•mC•fG•mG•fC•mU


263
1066
PmU•fG•mU•fC•mU•fC•mA•fG•mG•fC•mA•fG•mC•fC•mA•fC•mG•fG•mC


264
1067
PmU•fG•mG•fU•mC•fU•mC•fA•mG•fG•mC•fA•mG•fC•mC•fA•mC•fG•mG


265
1068
PmU•fA•mG•fG•mU•fC•mU•fC•mA•fG•mG•fC•mA•fG•mC•fC•mA•fC•mG


266
1069
PmU•fU•mG•fA•mG•fG•mU•fC•mU•fC•mA•fG•mG•fC•mA•fG•mC•fC•mA


267
1070
PmU•fU•mU•fG•mA•fG•mG•fU•mC•fU•mC•fA•mG•fG•mC•fA•mG•fC•mC


268
1071
PmU•fA•mU•fU•mG•fA•mG•fG•mU•fC•mU•fC•mA•fG•mG•fC•mA•fG•mC


269
1072
PmU•fU•mA•fU•mU•fG•mA•fG•mG•fU•mC•fU•mC•fA•mG•fG•mC•fA•mG


270
1073
PmU•fG•mU•fA•mU•fU•mG•fA•mG•fG•mU•fC•mU•fC•mA•fG•mG•fC•mA


271
1074
PmU•fG•mG•fU•mA•fU•mU•fG•mA•fG•mG•fU•mC•fU•mC•fA•mG•fG•mC


272
1075
PmU•fG•mG•fG•mU•fA•mU•fU•mG•fA•mG•fG•mU•fC•mU•fC•mA•fG•mG


273
1076
PmU•fU•mA•fG•mG•fC•mA•fG•mG•fU•mG•fG•mA•fC•mU•fU•mG•fG•mG


274
1077
PmU•fA•mU•fA•mG•fG•mC•fA•mG•fG•mU•fG•mG•fA•mC•fU•mU•fG•mG


275
1078
PmU•fG•mA•fU•mA•fG•mG•fC•mA•fG•mG•fU•mG•fG•mA•fC•mU•fU•mG


276
1079
PmU•fG•mG•fA•mU•fA•mG•fG•mC•fA•mG•fG•mU•fG•mG•fA•mC•fU•mU


277
1080
PmU•fU•mG•fG•mA•fU•mA•fG•mG•fC•mA•fG•mG•fU•mG•fG•mA•fC•mU


278
1081
PmU•fA•mU•fG•mG•fA•mU•fA•mG•fG•mC•fA•mG•fG•mU•fG•mG•fA•mC


279
1082
PmU•fG•mA•fU•mG•fG•mA•fU•mA•fG•mG•fC•mA•fG•mG•fU•mG•fG•mA


280
1083
PmU•fG•mG•fA•mU•fG•mG•fA•mU•fA•mG•fG•mC•fA•mG•fG•mU•fG•mG


281
1084
PmU•fA•mG•fG•mA•fU•mG•fG•mA•fU•mA•fG•mG•fC•mA•fG•mG•fU•mG


282
1085
PmU•fC•mA•fG•mG•fA•mU•fG•mG•fA•mU•fA•mG•fG•mC•fA•mG•fG•mU


283
1086
PmU•fG•mC•fA•mG•fG•mA•fU•mG•fG•mA•fU•mA•fG•mG•fC•mA•fG•mG


284
1087
PmU•fC•mG•fC•mA•fG•mG•fA•mU•fG•mG•fA•mU•fA•mG•fG•mC•fA•mG


285
1088
PmU•fU•mC•fG•mC•fA•mG•fG•mA•fU•mG•fG•mA•fU•mA•fG•mG•fC•mA


286
1089
PmU•fC•mU•fC•mG•fC•mA•fG•mG•fA•mU•fG•mG•fA•mU•fA•mG•fG•mC


287
1090
PmU•fG•mC•fU•mC•fG•mC•fA•mG•fG•mA•fU•mG•fG•mA•fU•mA•fG•mG


288
1091
PmU•fA•mG•fC•mU•fC•mG•fC•mA•fG•mG•fA•mU•fG•mG•fA•mU•fA•mG


289
1092
PmU•fG•mA•fG•mC•fU•mC•fG•mC•fA•mG•fG•mA•fU•mG•fG•mA•fU•mA


290
1093
PmU•fG•mG•fA•mG•fC•mU•fC•mG•fC•mA•fG•mG•fA•mU•fG•mG•fA•mU


291
1094
PmU•fA•mG•fG•mA•fG•mC•fU•mC•fG•mC•fA•mG•fG•mA•fU•mG•fG•mA


292
1095
PmU•fA•mA•fG•mG•fA•mG•fC•mU•fC•mG•fC•mA•fG•mG•fA•mU•fG•mG


293
1096
PmU•fC•mA•fA•mG•fG•mA•fG•mC•fU•mC•fG•mC•fA•mG•fG•mA•fU•mG


294
1097
PmU•fC•mC•fA•mA•fG•mG•fA•mG•fC•mU•fC•mG•fC•mA•fG•mG•fA•mU


295
1098
PmU•fC•mC•fC•mA•fA•mG•fG•mA•fG•mC•fU•mC•fG•mC•fA•mG•fG•mA


296
1099
PmU•fA•mC•fC•mC•fA•mA•fG•mG•fA•mG•fC•mU•fC•mG•fC•mA•fG•mG


297
1100
PmU•fG•mA•fC•mC•fC•mA•fA•mG•fG•mA•fG•mC•fU•mC•fG•mC•fA•mG


298
1101
PmU•fG•mG•fA•mC•fC•mC•fA•mA•fG•mG•fA•mG•fC•mU•fC•mG•fC•mA


299
1102
PmU•fA•mG•fG•mA•fC•mC•fC•mA•fA•mG•fG•mA•fG•mC•fU•mC•fG•mC


300
1103
PmU•fC•mA•fG•mG•fA•mC•fC•mC•fA•mA•fG•mG•fA•mG•fC•mU•fC•mG


301
1104
PmU•fG•mC•fA•mG•fG•mA•fC•mC•fC•mA•fA•mG•fG•mA•fG•mC•fU•mC


302
1105
PmU•fU•mG•fC•mA•fG•mG•fA•mC•fC•mC•fA•mA•fG•mG•fA•mG•fC•mU


303
1106
PmU•fU•mU•fG•mC•fA•mG•fG•mA•fC•mC•fC•mA•fA•mG•fG•mA•fG•mC


304
1107
PmU•fA•mU•fU•mG•fC•mA•fG•mG•fA•mC•fC•mC•fA•mA•fG•mG•fA•mG


305
1108
PmU•fG•mA•fU•mU•fG•mC•fA•mG•fG•mA•fC•mC•fC•mA•fA•mG•fG•mA


306
1109
PmU•fA•mG•fA•mU•fU•mG•fC•mA•fG•mG•fA•mC•fC•mC•fA•mA•fG•mG


307
1110
PmU•fG•mA•fG•mA•fU•mU•fG•mC•fA•mG•fG•mA•fC•mC•fC•mA•fA•mG


308
1111
PmU•fG•mG•fA•mG•fA•mU•fU•mG•fC•mA•fG•mG•fA•mC•fC•mC•fA•mA


309
1112
PmU•fU•mG•fG•mA•fG•mA•fU•mU•fG•mC•fA•mG•fG•mA•fC•mC•fC•mA


310
1113
PmU•fC•mU•fG•mG•fA•mG•fA•mU•fU•mG•fC•mA•fG•mG•fA•mC•fC•mC


311
1114
PmU•fC•mC•fU•mG•fG•mA•fG•mA•fU•mU•fG•mC•fA•mG•fG•mA•fC•mC


312
1115
PmU•fC•mC•fC•mU•fG•mG•fA•mG•fA•mU•fU•mG•fC•mA•fG•mG•fA•mC


313
1116
PmU•fG•mC•fC•mC•fU•mG•fG•mA•fG•mA•fU•mU•fG•mC•fA•mG•fG•mA


314
1117
PmU•fA•mG•fC•mC•fC•mU•fG•mG•fA•mG•fA•mU•fU•mG•fC•mA•fG•mG


315
1118
PmU•fC•mA•fG•mC•fC•mC•fU•mG•fG•mA•fG•mA•fU•mU•fG•mC•fA•mG


316
1119
PmU•fG•mC•fA•mG•fC•mC•fC•mU•fG•mG•fA•mG•fA•mU•fU•mG•fC•mA


317
1120
PmU•fG•mG•fC•mA•fG•mC•fC•mC•fU•mG•fG•mA•fG•mA•fU•mU•fG•mC


318
1121
PmU•fG•mG•fG•mC•fA•mG•fC•mC•fC•mU•fG•mG•fA•mG•fA•mU•fU•mG


319
1122
PmU•fU•mU•fU•mA•fA•mG•fC•mA•fA•mC•fC•mU•fA•mC•fA•mG•fG•mG


320
1123
PmU•fU•mU•fU•mU•fA•mA•fG•mC•fA•mA•fC•mC•fU•mA•fC•mA•fG•mG


321
1124
PmU•fC•mU•fU•mU•fU•mA•fA•mG•fC•mA•fA•mC•fC•mU•fA•mC•fA•mG


322
1125
PmU•fC•mC•fU•mU•fU•mU•fA•mA•fG•mC•fA•mA•fC•mC•fU•mA•fC•mA


323
1126
PmU•fC•mC•fC•mU•fU•mU•fU•mA•fA•mG•fC•mA•fA•mC•fC•mU•fA•mC


324
1127
PmU•fU•mC•fC•mC•fU•mU•fU•mU•fA•mA•fG•mC•fA•mA•fC•mC•fU•mA


325
1128
PmU•fG•mU•fC•mC•fC•mU•fU•mU•fU•mA•fA•mG•fC•mA•fA•mC•fC•mU


326
1129
PmU•fA•mC•fU•mG•fU•mC•fC•mC•fU•mU•fU•mU•fA•mA•fG•mC•fA•mA


327
1130
PmU•fU•mA•fC•mU•fG•mU•fC•mC•fC•mU•fU•mU•fU•mA•fA•mG•fC•mA


328
1131
PmU•fA•mU•fA•mC•fU•mG•fU•mC•fC•mC•fU•mU•fU•mU•fA•mA•fG•mC


329
1132
PmU•fA•mA•fU•mA•fC•mU•fG•mU•fC•mC•fC•mU•fU•mU•fU•mA•fA•mG


330
1133
PmU•fG•mA•fA•mU•fA•mC•fU•mG•fU•mC•fC•mC•fU•mU•fU•mU•fA•mA


331
1134
PmU•fA•mG•fA•mA•fU•mA•fC•mU•fG•mU•fC•mC•fC•mU•fU•mU•fU•mA


332
1135
PmU•fG•mA•fG•mA•fA•mU•fA•mC•fU•mG•fU•mC•fC•mC•fU•mU•fU•mU


333
1136
PmU•fU•mG•fA•mG•fA•mA•fU•mA•fC•mU•fG•mU•fC•mC•fC•mU•fU•mU


334
1137
PmU•fC•mU•fG•mA•fG•mA•fA•mU•fA•mC•fU•mG•fU•mC•fC•mC•fU•mU


335
1138
PmU•fA•mC•fU•mG•fA•mG•fA•mA•fU•mA•fC•mU•fG•mU•fC•mC•fC•mU


336
1139
PmU•fC•mA•fC•mU•fG•mA•fG•mA•fA•mU•fA•mC•fU•mG•fU•mC•fC•mC


337
1140
PmU•fG•mC•fA•mC•fU•mG•fA•mG•fA•mA•fU•mA•fC•mU•fG•mU•fC•mC


338
1141
PmU•fA•mG•fC•mA•fC•mU•fG•mA•fG•mA•fA•mU•fA•mC•fU•mG•fU•mC


339
1142
PmU•fG•mA•fG•mC•fA•mC•fU•mG•fA•mG•fA•mA•fU•mA•fC•mU•fG•mU


340
1143
PmU•fA•mG•fA•mG•fC•mA•fC•mU•fG•mA•fG•mA•fA•mU•fA•mC•fU•mG


341
1144
PmU•fG•mA•fG•mA•fG•mC•fA•mC•fU•mG•fA•mG•fA•mA•fU•mA•fC•mU


342
1145
PmU•fG•mG•fA•mG•fA•mG•fC•mA•fC•mU•fG•mA•fG•mA•fA•mU•fA•mC


343
1146
PmU•fA•mG•fG•mA•fG•mA•fG•mC•fA•mC•fU•mG•fA•mG•fA•mA•fU•mA


344
1147
PmU•fU•mA•fG•mG•fA•mG•fA•mG•fC•mA•fC•mU•fG•mA•fG•mA•fA•mU


345
1148
PmU•fG•mU•fA•mG•fG•mA•fG•mA•fG•mC•fA•mC•fU•mG•fA•mG•fA•mA


346
1149
PmU•fG•mG•fU•mA•fG•mG•fA•mG•fA•mG•fC•mA•fC•mU•fG•mA•fG•mA


347
1150
PmU•fG•mG•fG•mU•fA•mG•fG•mA•fG•mA•fG•mC•fA•mC•fU•mG•fA•mG


348
1151
PmU•fG•mG•fC•mC•fA•mG•fG•mC•fA•mU•fG•mA•fG•mG•fU•mG•fG•mG


349
1152
PmU•fG•mG•fG•mC•fC•mA•fG•mG•fC•mA•fU•mG•fA•mG•fG•mU•fG•mG


350
1153
PmU•fG•mC•fC•mA•fG•mC•fA•mU•fG•mC•fC•mU•fG•mG•fA•mG•fG•mG


351
1154
PmU•fG•mG•fC•mC•fA•mG•fC•mA•fU•mG•fC•mC•fU•mG•fG•mA•fG•mG


352
1155
PmU•fA•mG•fG•mC•fC•mA•fG•mC•fA•mU•fG•mC•fC•mU•fG•mG•fA•mG


353
1156
PmU•fG•mA•fG•mG•fC•mC•fA•mG•fC•mA•fU•mG•fC•mC•fU•mG•fG•mA


354
1157
PmU•fG•mG•fA•mG•fG•mC•fC•mA•fG•mC•fA•mU•fG•mC•fC•mU•fG•mG


355
1158
PmU•fG•mG•fG•mA•fG•mG•fC•mC•fA•mG•fC•mA•fU•mG•fC•mC•fU•mG


356
1159
PmU•fU•mG•fG•mG•fA•mG•fG•mC•fC•mA•fG•mC•fA•mU•fG•mC•fC•mU


357
1160
PmU•fA•mU•fU•mG•fG•mG•fA•mG•fG•mC•fC•mA•fG•mC•fA•mU•fG•mC


358
1161
PmU•fU•mA•fU•mU•fG•mG•fG•mA•fG•mG•fC•mC•fA•mG•fC•mA•fU•mG


359
1162
PmU•fU•mU•fA•mU•fU•mG•fG•mG•fA•mG•fG•mC•fC•mA•fG•mC•fA•mU


360
1163
PmU•fU•mU•fU•mA•fU•mU•fG•mG•fG•mA•fG•mG•fC•mC•fA•mG•fC•mA


361
1164
PmU•fC•mU•fU•mU•fA•mU•fU•mG•fG•mG•fA•mG•fG•mC•fC•mA•fG•mC


362
1165
PmU•fG•mC•fU•mU•fU•mA•fU•mU•fG•mG•fG•mA•fG•mG•fC•mC•fA•mG


363
1166
PmU•fA•mG•fC•mU•fU•mU•fA•mU•fU•mG•fG•mG•fA•mG•fG•mC•fC•mA


364
1167
PmU•fC•mA•fG•mC•fU•mU•fU•mA•fU•mU•fG•mG•fG•mA•fG•mG•fC•mC


365
1168
PmU•fC•mC•fA•mG•fC•mU•fU•mU•fA•mU•fU•mG•fG•mG•fA•mG•fG•mC


366
1169
PmU•fU•mC•fC•mA•fG•mC•fU•mU•fU•mA•fU•mU•fG•mG•fG•mA•fG•mG


367
1170
PmU•fG•mU•fC•mC•fA•mG•fC•mU•fU•mU•fA•mU•fU•mG•fG•mG•fA•mG


368
1171
PmU•fU•mU•fG•mU•fC•mC•fA•mG•fC•mU•fU•mU•fA•mU•fU•mG•fG•mG


369
1172
PmU•fC•mU•fU•mG•fU•mC•fC•mA•fG•mC•fU•mU•fU•mA•fU•mU•fG•mG


370
1173
PmU•fU•mC•fU•mU•fG•mU•fC•mC•fA•mG•fC•mU•fU•mU•fA•mU•fU•mG


371
1174
PmU•fU•mU•fC•mU•fU•mG•fU•mC•fC•mA•fG•mC•fU•mU•fU•mA•fU•mU


372
1175
PmU•fC•mU•fU•mC•fU•mU•fG•mU•fC•mC•fA•mG•fC•mU•fU•mU•fA•mU


373
1176
PmU•fG•mC•fU•mU•fC•mU•fU•mG•fU•mC•fC•mA•fG•mC•fU•mU•fU•mA


374
1177
PmU•fG•mC•fA•mG•fC•mU•fU•mC•fU•mU•fG•mU•fC•mC•fA•mG•fC•mU


375
1178
PmU•fU•mA•fG•mC•fA•mG•fC•mU•fU•mC•fU•mU•fG•mU•fC•mC•fA•mG


376
1179
PmU•fA•mU•fA•mG•fC•mA•fG•mC•fU•mU•fC•mU•fU•mG•fU•mC•fC•mA
















TABLE 1c







Nucleobase sequences of the sense


strands of 376 exemplary constructs










SEQ ID



#
NO:
Nucleobase sequence












1
401
AGUUCAUCCCUAGAA


 


2
402
GUUCAUCCCUAGAGA


 


3
403
UUCAUCCCUAGAGGA


 


4
404
UCAUCCCUAGAGGCA


 


5
405
CAUCCCUAGAGGCAA


 


6
406
AUCCCUAGAGGCAGA


 


7
407
UCCCUAGAGGCAGCA


 


8
408
CCCUAGAGGCAGCUA


 


9
409
CUAGAGGCAGCUGCA


 


10
410
UAGAGGCAGCUGCUA


 


11
411
AGAGGCAGCUGCUCA


 


12
412
GAGGCAGCUGCUCCA


 


13
413
CUGCUCCAGGAACAA


 


14
414
UGCUCCAGGAACAGA


 


15
415
UCCAGGAACAGAGGA


 


16
416
CCAGGAACAGAGGUA


 


17
417
CAGGAACAGAGGUGA


 


18
418
AGGAACAGAGGUGCA


 


19
419
GGAACAGAGGUGCCA


 


20
420
GAACAGAGGUGCCAA


 


21
421
AACAGAGGUGCCAUA


 


22
422
ACAGAGGUGCCAUGA


 


23
423
AGAGGUGCCAUGCAA


 


24
424
GAGGUGCCAUGCAGA


 


25
425
AGGUGCCAUGCAGCA


 


26
426
GGUGCCAUGCAGCCA


 


27
427
GUGCCAUGCAGCCCA


 


28
428
GGUACUCCUUGUUGA


 


29
429
GUACUCCUUGUUGUA


 


30
430
UACUCCUUGUUGUUA


 


31
431
ACUCCUUGUUGUUGA


 


32
432
CUCCUUGUUGUUGCA


 


33
433
UCCUUGUUGUUGCCA


 


34
434
CCUUGUUGUUGCCCA


 


35
435
CUUGUUGUUGCCCUA


 


36
436
UUGUUGUUGCCCUCA


 


37
437
UGUUGUUGCCCUCCA


 


38
438
GUUGUUGCCCUCCUA


 


39
439
UUGUUGCCCUCCUGA


 


40
440
UGUUGCCCUCCUGGA


 


41
441
GUUGCCCUCCUGGCA


 


42
442
UUGCCCUCCUGGCGA


 


43
443
UGCCCUCCUGGCGCA


 


44
444
GCCCUCCUGGCGCUA


 


45
445
CCCUCCUGGCGCUCA


 


46
446
CCUCCUGGCGCUCCA


 


47
447
CUCCUGGCGCUCCUA


 


48
448
CUGGCGCUCCUGGCA


 


49
449
CGCUCCUGGCCUCUA


 


50
450
GCUCCUGGCCUCUGA


 


51
451
CUCCUGGCCUCUGCA


 


52
452
UCCUGGCCUCUGCCA


 


53
453
CCUGGCCUCUGCCCA


 


54
454
UGGCCUCUGCCCGAA


 


55
455
GGCCUCUGCCCGAGA


 


56
456
GCCUCUGCCCGAGCA


 


57
457
CCUCUGCCCGAGCUA


 


58
458
CUCUGCCCGAGCUUA


 


59
459
UCUGCCCGAGCUUCA


 


60
460
CUGCCCGAGCUUCAA


 


61
461
UGCCCGAGCUUCAGA


 


62
462
GCCCGAGCUUCAGAA


 


63
463
CCCGAGCUUCAGAGA


 


64
464
CCGAGCUUCAGAGGA


 


65
465
CGAGCUUCAGAGGCA


 


66
466
GAGCUUCAGAGGCCA


 


67
467
AGCUUCAGAGGCCGA


 


68
468
GCUUCAGAGGCCGAA


 


69
469
CUUCAGAGGCCGAGA


 


70
470
UUCAGAGGCCGAGGA


 


71
471
UCAGAGGCCGAGGAA


 


72
472
CAGAGGCCGAGGAUA


 


73
473
AGAGGCCGAGGAUGA


 


74
474
GAGGCCGAGGAUGCA


 


75
475
AGGCCGAGGAUGCCA


 


76
476
GGCCGAGGAUGCCUA


 


77
477
GCCGAGGAUGCCUCA


 


78
478
CCGAGGAUGCCUCCA


 


79
479
CGAGGAUGCCUCCCA


 


80
480
GAGGAUGCCUCCCUA


 


81
481
AGGAUGCCUCCCUUA


 


82
482
GGAUGCCUCCCUUCA


 


83
483
GAUGCCUCCCUUCUA


 


84
484
AUGCCUCCCUUCUCA


 


85
485
UGCCUCCCUUCUCAA


 


86
486
GCCUCCCUUCUCAGA


 


87
487
CCUCCCUUCUCAGCA


 


88
488
CUCCCUUCUCAGCUA


 


89
489
CCUUCUCAGCUUCAA


 


90
490
CUUCUCAGCUUCAUA


 


91
491
UUCUCAGCUUCAUGA


 


92
492
UCUCAGCUUCAUGCA


 


93
493
CUCAGCUUCAUGCAA


 


94
494
UCAGCUUCAUGCAGA


 


95
495
CAGCUUCAUGCAGGA


 


96
496
AGCUUCAUGCAGGGA


 


97
497
GCUUCAUGCAGGGUA


 


98
498
CUUCAUGCAGGGUUA


 


99
499
UUCAUGCAGGGUUAA


 


100
500
UCAUGCAGGGUUACA


 


101
501
CAUGCAGGGUUACAA


 


102
502
AUGCAGGGUUACAUA


 


103
503
UGCAGGGUUACAUGA


 


104
504
GCAGGGUUACAUGAA


 


105
505
CAGGGUUACAUGAAA


 


106
506
AGGGUUACAUGAAGA


 


107
507
GGGUUACAUGAAGCA


 


108
508
GGUUACAUGAAGCAA


 


109
509
GUUACAUGAAGCACA


 


110
510
UUACAUGAAGCACGA


 


111
511
UACAUGAAGCACGCA


 


112
512
ACAUGAAGCACGCCA


 


113
513
CAUGAAGCACGCCAA


 


114
514
AUGAAGCACGCCACA


 


115
515
UGAAGCACGCCACCA


 


116
516
GAAGCACGCCACCAA


 


117
517
AAGCACGCCACCAAA


 


118
518
AGCACGCCACCAAGA


 


119
519
GCACGCCACCAAGAA


 


120
520
CACGCCACCAAGACA


 


121
521
ACGCCACCAAGACCA


 


122
522
CGCCACCAAGACCGA


 


123
523
GCCACCAAGACCGCA


 


124
524
CCACCAAGACCGCCA


 


125
525
CACCAAGACCGCCAA


 


126
526
ACCAAGACCGCCAAA


 


127
527
CCAAGACCGCCAAGA


 


128
528
CAAGACCGCCAAGGA


 


129
529
AAGACCGCCAAGGAA


 


130
530
AGACCGCCAAGGAUA


 


131
531
GACCGCCAAGGAUGA


 


132
532
ACCGCCAAGGAUGCA


 


133
533
CCGCCAAGGAUGCAA


 


134
534
CGCCAAGGAUGCACA


 


135
535
GCCAAGGAUGCACUA


 


136
536
CCAAGGAUGCACUGA


 


137
537
CAAGGAUGCACUGAA


 


138
538
AAGGAUGCACUGAGA


 


139
539
AGGAUGCACUGAGCA


 


140
540
GGAUGCACUGAGCAA


 


141
541
GAUGCACUGAGCAGA


 


142
542
AUGCACUGAGCAGCA


 


143
543
UGCACUGAGCAGCGA


 


144
544
GCACUGAGCAGCGUA


 


145
545
CACUGAGCAGCGUGA


 


146
546
ACUGAGCAGCGUGCA


 


147
547
CUGAGCAGCGUGCAA


 


148
548
UGAGCAGCGUGCAGA


 


149
549
GAGCAGCGUGCAGGA


 


150
550
AGCAGCGUGCAGGAA


 


151
551
CAGCGUGCAGGAGUA


 


152
552
CGUGCAGGAGUCCCA


 


153
553
GUGCAGGAGUCCCAA


 


154
554
UGCAGGAGUCCCAGA


 


155
555
GCAGGAGUCCCAGGA


 


156
556
CAGGAGUCCCAGGUA


 


157
557
AGGAGUCCCAGGUGA


 


158
558
GGAGUCCCAGGUGGA


 


159
559
GUCCCAGGUGGCCCA


 


160
560
UCCCAGGUGGCCCAA


 


161
561
CAGGUGGCCCAGCAA


 


162
562
AGGUGGCCCAGCAGA


 


163
563
UGGCCCAGCAGGCCA


 


164
564
CCCAGCAGGCCAGGA


 


165
565
UGGGUGACCGAUGGA


 


166
566
GGGUGACCGAUGGCA


 


167
567
GGUGACCGAUGGCUA


 


168
568
GUGACCGAUGGCUUA


 


169
569
UGACCGAUGGCUUCA


 


170
570
GACCGAUGGCUUCAA


 


171
571
ACCGAUGGCUUCAGA


 


172
572
CCGAUGGCUUCAGUA


 


173
573
CGAUGGCUUCAGUUA


 


174
574
GAUGGCUUCAGUUCA


 


175
575
AUGGCUUCAGUUCCA


 


176
576
UGGCUUCAGUUCCCA


 


177
577
GGCUUCAGUUCCCUA


 


178
578
GCUUCAGUUCCCUGA


 


179
579
CUUCAGUUCCCUGAA


 


180
580
UUCAGUUCCCUGAAA


 


181
581
UCAGUUCCCUGAAAA


 


182
582
CAGUUCCCUGAAAGA


 


183
583
AGUUCCCUGAAAGAA


 


184
584
GUUCCCUGAAAGACA


 


185
585
UUCCCUGAAAGACUA


 


186
586
UCCCUGAAAGACUAA


 


187
587
CCCUGAAAGACUACA


 


188
588
CCUGAAAGACUACUA


 


189
589
CUGAAAGACUACUGA


 


190
590
UGAAAGACUACUGGA


 


191
591
GAAAGACUACUGGAA


 


192
592
AAAGACUACUGGAGA


 


193
593
AAGACUACUGGAGCA


 


194
594
AGACUACUGGAGCAA


 


195
595
GACUACUGGAGCACA


 


196
596
ACUACUGGAGCACCA


 


197
597
CUACUGGAGCACCGA


 


198
598
UACUGGAGCACCGUA


 


199
599
ACUGGAGCACCGUUA


 


200
600
CUGGAGCACCGUUAA


 


201
601
UGGAGCACCGUUAAA


 


202
602
GGAGCACCGUUAAGA


 


203
603
GAGCACCGUUAAGGA


 


204
604
AGCACCGUUAAGGAA


 


205
605
GCACCGUUAAGGACA


 


206
606
CACCGUUAAGGACAA


 


207
607
ACCGUUAAGGACAAA


 


208
608
CCGUUAAGGACAAGA


 


209
609
CGUUAAGGACAAGUA


 


210
610
GUUAAGGACAAGUUA


 


211
611
UUAAGGACAAGUUCA


 


212
612
UAAGGACAAGUUCUA


 


213
613
AAGGACAAGUUCUCA


 


214
614
AGGACAAGUUCUCUA


 


215
615
GGACAAGUUCUCUGA


 


216
616
GACAAGUUCUCUGAA


 


217
617
ACAAGUUCUCUGAGA


 


218
618
CAAGUUCUCUGAGUA


 


219
619
AAGUUCUCUGAGUUA


 


220
620
AGUUCUCUGAGUUCA


 


221
621
UUCUCUGAGUUCUGA


 


222
622
UCUCUGAGUUCUGGA


 


223
623
CUCUGAGUUCUGGGA


 


224
624
UCUGAGUUCUGGGAA


 


225
625
CUGAGUUCUGGGAUA


 


226
626
UGAGUUCUGGGAUUA


 


227
627
GAGUUCUGGGAUUUA


 


228
628
AGUUCUGGGAUUUGA


 


229
629
GUUCUGGGAUUUGGA


 


230
630
UUCUGGGAUUUGGAA


 


231
631
UCUGGGAUUUGGACA


 


232
632
CUGGGAUUUGGACCA


 


233
633
UGGGAUUUGGACCCA


 


234
634
GGGAUUUGGACCCUA


 


235
635
GGAUUUGGACCCUGA


 


236
636
GAUUUGGACCCUGAA


 


237
637
UGGACCCUGAGGUCA


 


238
638
GGACCCUGAGGUCAA


 


239
639
GACCCUGAGGUCAGA


 


240
640
ACCCUGAGGUCAGAA


 


241
641
CCCUGAGGUCAGACA


 


242
642
CCUGAGGUCAGACCA


 


243
643
CUGAGGUCAGACCAA


 


244
644
UGAGGUCAGACCAAA


 


245
645
GAGGUCAGACCAACA


 


246
646
AGGUCAGACCAACUA


 


247
647
GGUCAGACCAACUUA


 


248
648
GUCAGACCAACUUCA


 


249
649
UCAGACCAACUUCAA


 


250
650
CAGACCAACUUCAGA


 


251
651
GACCAACUUCAGCCA


 


252
652
ACCAACUUCAGCCGA


 


253
653
CCAACUUCAGCCGUA


 


254
654
CAACUUCAGCCGUGA


 


255
655
AACUUCAGCCGUGGA


 


256
656
ACUUCAGCCGUGGCA


 


257
657
UUCAGCCGUGGCUGA


 


258
658
UCAGCCGUGGCUGCA


 


259
659
CAGCCGUGGCUGCCA


 


260
660
AGCCGUGGCUGCCUA


 


261
661
GCCGUGGCUGCCUGA


 


262
662
GUGGCUGCCUGAGAA


 


263
663
UGGCUGCCUGAGACA


 


264
664
GGCUGCCUGAGACCA


 


265
665
GCUGCCUGAGACCUA


 


266
666
UGCCUGAGACCUCAA


 


267
667
GCCUGAGACCUCAAA


 


268
668
CCUGAGACCUCAAUA


 


269
669
CUGAGACCUCAAUAA


 


270
670
UGAGACCUCAAUACA


 


271
671
GAGACCUCAAUACCA


 


272
672
AGACCUCAAUACCCA


 


273
673
AGUCCACCUGCCUAA


 


274
674
GUCCACCUGCCUAUA


 


275
675
UCCACCUGCCUAUCA


 


276
676
CCACCUGCCUAUCCA


 


277
677
CACCUGCCUAUCCAA


 


278
678
ACCUGCCUAUCCAUA


 


279
679
CCUGCCUAUCCAUCA


 


280
680
CUGCCUAUCCAUCCA


 


281
681
UGCCUAUCCAUCCUA


 


282
682
GCCUAUCCAUCCUGA


 


283
683
CCUAUCCAUCCUGCA


 


284
684
CUAUCCAUCCUGCGA


 


285
685
UAUCCAUCCUGCGAA


 


286
686
AUCCAUCCUGCGAGA


 


287
687
UCCAUCCUGCGAGCA


 


288
688
CCAUCCUGCGAGCUA


 


289
689
CAUCCUGCGAGCUCA


 


290
690
AUCCUGCGAGCUCCA


 


291
691
UCCUGCGAGCUCCUA


 


292
692
CCUGCGAGCUCCUUA


 


293
693
CUGCGAGCUCCUUGA


 


294
694
UGCGAGCUCCUUGGA


 


295
695
GCGAGCUCCUUGGGA


 


296
696
CGAGCUCCUUGGGUA


 


297
697
GAGCUCCUUGGGUCA


 


298
698
AGCUCCUUGGGUCCA


 


299
699
GCUCCUUGGGUCCUA


 


300
700
CUCCUUGGGUCCUGA


 


301
701
UCCUUGGGUCCUGCA


 


302
702
CCUUGGGUCCUGCAA


 


303
703
CUUGGGUCCUGCAAA


 


304
704
UUGGGUCCUGCAAUA


 


305
705
UGGGUCCUGCAAUCA


 


306
706
GGGUCCUGCAAUCUA


 


307
707
GGUCCUGCAAUCUCA


 


308
708
GUCCUGCAAUCUCCA


 


309
709
UCCUGCAAUCUCCAA


 


310
710
CCUGCAAUCUCCAGA


 


311
711
CUGCAAUCUCCAGGA


 


312
712
UGCAAUCUCCAGGGA


 


313
713
GCAAUCUCCAGGGCA


 


314
714
CAAUCUCCAGGGCUA


 


315
715
AAUCUCCAGGGCUGA


 


316
716
AUCUCCAGGGCUGCA


 


317
717
UCUCCAGGGCUGCCA


 


318
718
CUCCAGGGCUGCCCA


 


319
719
GUAGGUUGCUUAAAA


 


320
720
UAGGUUGCUUAAAAA


 


321
721
AGGUUGCUUAAAAGA


 


322
722
GGUUGCUUAAAAGGA


 


323
723
GUUGCUUAAAAGGGA


 


324
724
UUGCUUAAAAGGGAA


 


325
725
UGCUUAAAAGGGACA


 


326
726
UUAAAAGGGACAGUA


 


327
727
UAAAAGGGACAGUAA


 


328
728
AAAAGGGACAGUAUA


 


329
729
AAAGGGACAGUAUUA


 


330
730
AAGGGACAGUAUUCA


 


331
731
AGGGACAGUAUUCUA


 


332
732
GGGACAGUAUUCUCA


 


333
733
GGACAGUAUUCUCAA


 


334
734
GACAGUAUUCUCAGA


 


335
735
ACAGUAUUCUCAGUA


 


336
736
CAGUAUUCUCAGUGA


 


337
737
AGUAUUCUCAGUGCA


 


338
738
GUAUUCUCAGUGCUA


 


339
739
UAUUCUCAGUGCUCA


 


340
740
AUUCUCAGUGCUCUA


 


341
741
UUCUCAGUGCUCUCA


 


342
742
UCUCAGUGCUCUCCA


 


343
743
CUCAGUGCUCUCCUA


 


344
744
UCAGUGCUCUCCUAA


 


345
745
CAGUGCUCUCCUACA


 


346
746
AGUGCUCUCCUACCA


 


347
747
GUGCUCUCCUACCCA


 


348
748
CCUCAUGCCUGGCCA


 


349
749
CUCAUGCCUGGCCCA


 


350
750
CCAGGCAUGCUGGCA


 


351
751
CAGGCAUGCUGGCCA


 


352
752
AGGCAUGCUGGCCUA


 


353
753
GGCAUGCUGGCCUCA


 


354
754
GCAUGCUGGCCUCCA


 


355
755
CAUGCUGGCCUCCCA


 


356
756
AUGCUGGCCUCCCAA


 


357
757
GCUGGCCUCCCAAUA


 


358
758
CUGGCCUCCCAAUAA


 


359
759
UGGCCUCCCAAUAAA


 


360
760
GGCCUCCCAAUAAAA


 


361
761
GCCUCCCAAUAAAGA


 


362
762
CCUCCCAAUAAAGCA


 


363
763
CUCCCAAUAAAGCUA


 


364
764
UCCCAAUAAAGCUGA


 


365
765
CCCAAUAAAGCUGGA


 


366
766
CCAAUAAAGCUGGAA


 


367
767
CAAUAAAGCUGGACA


 


368
768
AUAAAGCUGGACAAA


 


369
769
UAAAGCUGGACAAGA


 


370
770
AAAGCUGGACAAGAA


 


371
771
AAGCUGGACAAGAAA


 


372
772
AGCUGGACAAGAAGA


 


373
773
GCUGGACAAGAAGCA


 


374
774
GGACAAGAAGCUGCA


 


375
775
ACAAGAAGCUGCUAA


 


376
776
CAAGAAGCUGCUAUA
















TABLE 1d







Nucleobase sequences and sugar-phosphate backbone 


modifications of the sense strands of 376 exemplary constructs:










SEQ




ID



#
NO:
Nucleobase sequence and backbone modification





 1
1180
fA•mG•fU•mU•fC•mA•fU•mC•fC•mC•fU•mA•fG•mA•fA


 2
1181
fG•mU•fU•mC•fA•mU•fC•mC•fC•mU•fA•mG•fA•mG•fA


 3
1182
fU•mU•fC•mA•fU•mC•fC•mC•fU•mA•fG•mA•fG•mG•fA


 4
1183
fU•mC•fA•mU•fC•mC•fC•mU•fA•mG•fA•mG•fG•mC•fA


 5
1184
fC•mA•fU•mC•fC•mC•fU•mA•fG•mA•fG•mG•fC•mA•fA


 6
1185
fA•mU•fC•mC•fC•mU•fA•mG•fA•mG•fG•mC•fA•mG•fA


 7
1186
fU•mC•fC•mC•fU•mA•fG•mA•fG•mG•fC•mA•fG•mC•fA


 8
1187
fC•mC•fC•mU•fA•mG•fA•mG•fG•mC•fA•mG•fC•mU•fA


 9
1188
fC•mU•fA•mG•fA•mG•fG•mC•fA•mG•fC•mU•fG•mC•fA


 10
1189
fU•mA•fG•mA•fG•mG•fC•mA•fG•mC•fU•mG•fC•mU•fA


 11
1190
fA•mG•fA•mG•fG•mC•fA•mG•fC•mU•fG•mC•fU•mC•fA


 12
1191
fG•mA•fG•mG•fC•mA•fG•mC•fU•mG•fC•mU•fC•mC•fA


 13
1192
fC•mU•fG•mC•fU•mC•fC•mA•fG•mG•fA•mA•fC•mA•fA


 14
1193
fU•mG•fC•mU•fC•mC•fA•mG•fG•mA•fA•mC•fA•mG•fA


 15
1194
fU•mC•fC•mA•fG•mG•fA•mA•fC•mA•fG•mA•fG•mG•fA


 16
1195
fC•mC•fA•mG•fG•mA•fA•mC•fA•mG•fA•mG•fG•mU•fA


 17
1196
fC•mA•fG•mG•fA•mA•fC•mA•fG•mA•fG•mG•fU•mG•fA


 18
1197
fA•mG•fG•mA•fA•mC•fA•mG•fA•mG•fG•mU•fG•mC•fA


 19
1198
fG•mG•fA•mA•fC•mA•fG•mA•fG•mG•fU•mG•fC•mC•fA


 20
1199
fG•mA•fA•mC•fA•mG•fA•mG•fG•mU•fG•mC•fC•mA•fA


 21
1200
fA•mA•fC•mA•fG•mA•fG•mG•fU•mG•fC•mC•fA•mU•fA


 22
1201
fA•mC•fA•mG•fA•mG•fG•mU•fG•mC•fC•mA•fU•mG•fA


 23
1202
fA•mG•fA•mG•fG•mU•fG•mC•fC•mA•fU•mG•fC•mA•fA


 24
1203
fG•mA•fG•mG•fU•mG•fC•mC•fA•mU•fG•mC•fA•mG•fA


 25
1204
fA•mG•fG•mU•fG•mC•fC•mA•fU•mG•fC•mA•fG•mC•fA


 26
1205
fG•mG•fU•mG•fC•mC•fA•mU•fG•mC•fA•mG•fC•mC•fA


 27
1206
fG•mU•fG•mC•fC•mA•fU•mG•fC•mA•fG•mC•fC•mC•fA


 28
1207
fG•mG•fU•mA•fC•mU•fC•mC•fU•mU•fG•mU•fU•mG•fA


 29
1208
fG•mU•fA•mC•fU•mC•fC•mU•fU•mG•fU•mU•fG•mU•fA


 30
1209
fU•mA•fC•mU•fC•mC•fU•mU•fG•mU•fU•mG•fU•mU•fA


 31
1210
fA•mC•fU•mC•fC•mU•fU•mG•fU•mU•fG•mU•fU•mG•fA


 32
1211
fC•mU•fC•mC•fU•mU•fG•mU•fU•mG•fU•mU•fG•mC•fA


 33
1212
fU•mC•fC•mU•fU•mG•fU•mU•fG•mU•fU•mG•fC•mC•fA


 34
1213
fC•mC•fU•mU•fG•mU•fU•mG•fU•mU•fG•mC•fC•mC•fA


 35
1214
fC•mU•fU•mG•fU•mU•fG•mU•fU•mG•fC•mC•fC•mU•fA


 36
1215
fU•mU•fG•mU•fU•mG•fU•mU•fG•mC•fC•mC•fU•mC•fA


 37
1216
fU•mG•fU•mU•fG•mU•fU•mG•fC•mC•fC•mU•fC•mC•fA


 38
1217
fG•mU•fU•mG•fU•mU•fG•mC•fC•mC•fU•mC•fC•mU•fA


 39
1218
fU•mU•fG•mU•fU•mG•fC•mC•fC•mU•fC•mC•fU•mG•fA


 40
1219
fU•mG•fU•mU•fG•mC•fC•mC•fU•mC•fC•mU•fG•mG•fA


 41
1220
fG•mU•fU•mG•fC•mC•fC•mU•fC•mC•fU•mG•fG•mC•fA


 42
1221
fU•mU•fG•mC•fC•mC•fU•mC•fC•mU•fG•mG•fC•mG•fA


 43
1222
fU•mG•fC•mC•fC•mU•fC•mC•fU•mG•fG•mC•fG•mC•fA


 44
1223
fG•mC•fC•mC•fU•mC•fC•mU•fG•mG•fC•mG•fC•mU•fA


 45
1224
fC•mC•fC•mU•fC•mC•fU•mG•fG•mC•fG•mC•fU•mC•fA


 46
1225
fC•mC•fU•mC•fC•mU•fG•mG•fC•mG•fC•mU•fC•mC•fA


 47
1226
fC•mU•fC•mC•fU•mG•fG•mC•fG•mC•fU•mC•fC•mU•fA


 48
1227
fC•mU•fG•mG•fC•mG•fC•mU•fC•mC•fU•mG•fG•mC•fA


 49
1228
fC•mG•fC•mU•fC•mC•fU•mG•fG•mC•fC•mU•fC•mU•fA


 50
1229
fG•mC•fU•mC•fC•mU•fG•mG•fC•mC•fU•mC•fU•mG•fA


 51
1230
fC•mU•fC•mC•fU•mG•fG•mC•fC•mU•fC•mU•fG•mC•fA


 52
1231
fU•mC•fC•mU•fG•mG•fC•mC•fU•mC•fU•mG•fC•mC•fA


 53
1232
fC•mC•fU•mG•fG•mC•fC•mU•fC•mU•fG•mC•fC•mC•fA


 54
1233
fU•mG•fG•mC•fC•mU•fC•mU•fG•mC•fC•mC•fG•mA•fA


 55
1234
fG•mG•fC•mC•fU•mC•fU•mG•fC•mC•fC•mG•fA•mG•fA


 56
1235
fG•mC•fC•mU•fC•mU•fG•mC•fC•mC•fG•mA•fG•mC•fA


 57
1236
fC•mC•fU•mC•fU•mG•fC•mC•fC•mG•fA•mG•fC•mU•fA


 58
1237
fC•mU•fC•mU•fG•mC•fC•mC•fG•mA•fG•mC•fU•mU•fA


 59
1238
fU•mC•fU•mG•fC•mC•fC•mG•fA•mG•fC•mU•fU•mC•fA


 60
1239
fC•mU•fG•mC•fC•mC•fG•mA•fG•mC•fU•mU•fC•mA•fA


 61
1240
fU•mG•fC•mC•fC•mG•fA•mG•fC•mU•fU•mC•fA•mG•fA


 62
1241
fG•mC•fC•mC•fG•mA•fG•mC•fU•mU•fC•mA•fG•mA•fA


 63
1242
fC•mC•fC•mG•fA•mG•fC•mU•fU•mC•fA•mG•fA•mG•fA


 64
1243
fC•mC•fG•mA•fG•mC•fU•mU•fC•mA•fG•mA•fG•mG•fA


 65
1244
fC•mG•fA•mG•fC•mU•fU•mC•fA•mG•fA•mG•fG•mC•fA


 66
1245
fG•mA•fG•mC•fU•mU•fC•mA•fG•mA•fG•mG•fC•mC•fA


 67
1246
fA•mG•fC•mU•fU•mC•fA•mG•fA•mG•fG•mC•fC•mG•fA


 68
1247
fG•mC•fU•mU•fC•mA•fG•mA•fG•mG•fC•mC•fG•mA•fA


 69
1248
fC•mU•fU•mC•fA•mG•fA•mG•fG•mC•fC•mG•fA•mG•fA


 70
1249
fU•mU•fC•mA•fG•mA•fG•mG•fC•mC•fG•mA•fG•mG•fA


 71
1250
fU•mC•fA•mG•fA•mG•fG•mC•fC•mG•fA•mG•fG•mA•fA


 72
1251
fC•mA•fG•mA•fG•mG•fC•mC•fG•mA•fG•mG•fA•mU•fA


 73
1252
fA•mG•fA•mG•fG•mC•fC•mG•fA•mG•fG•mA•fU•mG•fA


 74
1253
fG•mA•fG•mG•fC•mC•fG•mA•fG•mG•fA•mU•fG•mC•fA


 75
1254
fA•mG•fG•mC•fC•mG•fA•mG•fG•mA•fU•mG•fC•mC•fA


 76
1255
fG•mG•fC•mC•fG•mA•fG•mG•fA•mU•fG•mC•fC•mU•fA


 77
1256
fG•mC•fC•mG•fA•mG•fG•mA•fU•mG•fC•mC•fU•mC•fA


 78
1257
fC•mC•fG•mA•fG•mG•fA•mU•fG•mC•fC•mU•fC•mC•fA


 79
1258
fC•mG•fA•mG•fG•mA•fU•mG•fC•mC•fU•mC•fC•mC•fA


 80
1259
fG•mA•fG•mG•fA•mU•fG•mC•fC•mU•fC•mC•fC•mU•fA


 81
1260
fA•mG•fG•mA•fU•mG•fC•mC•fU•mC•fC•mC•fU•mU•fA


 82
1261
fG•mG•fA•mU•fG•mC•fC•mU•fC•mC•fC•mU•fU•mC•fA


 83
1262
fG•mA•fU•mG•fC•mC•fU•mC•fC•mC•fU•mU•fC•mU•fA


 84
1263
fA•mU•fG•mC•fC•mU•fC•mC•fC•mU•fU•mC•fU•mC•fA


 85
1264
fU•mG•fC•mC•fU•mC•fC•mC•fU•mU•fC•mU•fC•mA•fA


 86
1265
fG•mC•fC•mU•fC•mC•fC•mU•fU•mC•fU•mC•fA•mG•fA


 87
1266
fC•mC•fU•mC•fC•mC•fU•mU•fC•mU•fC•mA•fG•mC•fA


 88
1267
fC•mU•fC•mC•fC•mU•fU•mC•fU•mC•fA•mG•fC•mU•fA


 89
1268
fC•mC•fU•mU•fC•mU•fC•mA•fG•mC•fU•mU•fC•mA•fA


 90
1269
fC•mU•fU•mC•fU•mC•fA•mG•fC•mU•fU•mC•fA•mU•fA


 91
1270
fU•mU•fC•mU•fC•mA•fG•mC•fU•mU•fC•mA•fU•mG•fA


 92
1271
fU•mC•fU•mC•fA•mG•fC•mU•fU•mC•fA•mU•fG•mC•fA


 93
1272
fC•mU•fC•mA•fG•mC•fU•mU•fC•mA•fU•mG•fC•mA•fA


 94
1273
fU•mC•fA•mG•fC•mU•fU•mC•fA•mU•fG•mC•fA•mG•fA


 95
1274
fC•mA•fG•mC•fU•mU•fC•mA•fU•mG•fC•mA•fG•mG•fA


 96
1275
fA•mG•fC•mU•fU•mC•fA•mU•fG•mC•fA•mG•fG•mG•fA


 97
1276
fG•mC•fU•mU•fC•mA•fU•mG•fC•mA•fG•mG•fG•mU•fA


 98
1277
fC•mU•fU•mC•fA•mU•fG•mC•fA•mG•fG•mG•fU•mU•fA


 99
1278
fU•mU•fC•mA•fU•mG•fC•mA•fG•mG•fG•mU•fU•mA•fA


100
1279
fU•mC•fA•mU•fG•mC•fA•mG•fG•mG•fU•mU•fA•mC•fA


101
1280
fC•mA•fU•mG•fC•mA•fG•mG•fG•mU•fU•mA•fC•mA•fA


102
1281
fA•mU•fG•mC•fA•mG•fG•mG•fU•mU•fA•mC•fA•mU•fA


103
1282
fU•mG•fC•mA•fG•mG•fG•mU•fU•mA•fC•mA•fU•mG•fA


104
1283
fG•mC•fA•mG•fG•mG•fU•mU•fA•mC•fA•mU•fG•mA•fA


105
1284
fC•mA•fG•mG•fG•mU•fU•mA•fC•mA•fU•mG•fA•mA•fA


106
1285
fA•mG•fG•mG•fU•mU•fA•mC•fA•mU•fG•mA•fA•mG•fA


107
1286
fG•mG•fG•mU•fU•mA•fC•mA•fU•mG•fA•mA•fG•mC•fA


108
1287
fG•mG•fU•mU•fA•mC•fA•mU•fG•mA•fA•mG•fC•mA•fA


109
1288
fG•mU•fU•mA•fC•mA•fU•mG•fA•mA•fG•mC•fA•mC•fA


110
1289
fU•mU•fA•mC•fA•mU•fG•mA•fA•mG•fC•mA•fC•mG•fA


111
1290
fU•mA•fC•mA•fU•mG•fA•mA•fG•mC•fA•mC•fG•mC•fA


112
1291
fA•mC•fA•mU•fG•mA•fA•mG•fC•mA•fC•mG•fC•mC•fA


113
1292
fC•mA•fU•mG•fA•mA•fG•mC•fA•mC•fG•mC•fC•mA•fA


114
1293
fA•mU•fG•mA•fA•mG•fC•mA•fC•mG•fC•mC•fA•mC•fA


115
1294
fU•mG•fA•mA•fG•mC•fA•mC•fG•mC•fC•mA•fC•mC•fA


116
1295
fG•mA•fA•mG•fC•mA•fC•mG•fC•mC•fA•mC•fC•mA•fA


117
1296
fA•mA•fG•mC•fA•mC•fG•mC•fC•mA•fC•mC•fA•mA•fA


118
1297
fA•mG•fC•mA•fC•mG•fC•mC•fA•mC•fC•mA•fA•mG•fA


119
1298
fG•mC•fA•mC•fG•mC•fC•mA•fC•mC•fA•mA•fG•mA•fA


120
1299
fC•mA•fC•mG•fC•mC•fA•mC•fC•mA•fA•mG•fA•mC•fA


121
1300
fA•mC•fG•mC•fC•mA•fC•mC•fA•mA•fG•mA•fC•mC•fA


122
1301
fC•mG•fC•mC•fA•mC•fC•mA•fA•mG•fA•mC•fC•mG•fA


123
1302
fG•mC•fC•mA•fC•mC•fA•mA•fG•mA•fC•mC•fG•mC•fA


124
1303
fC•mC•fA•mC•fC•mA•fA•mG•fA•mC•fC•mG•fC•mC•fA


125
1304
fC•mA•fC•mC•fA•mA•fG•mA•fC•mC•fG•mC•fC•mA•fA


126
1305
fA•mC•fC•mA•fA•mG•fA•mC•fC•mG•fC•mC•fA•mA•fA


127
1306
fC•mC•fA•mA•fG•mA•fC•mC•fG•mC•fC•mA•fA•mG•fA


128
1307
fC•mA•fA•mG•fA•mC•fC•mG•fC•mC•fA•mA•fG•mG•fA


129
1308
fA•mA•fG•mA•fC•mC•fG•mC•fC•mA•fA•mG•fG•mA•fA


130
1309
fA•mG•fA•mC•fC•mG•fC•mC•fA•mA•fG•mG•fA•mU•fA


131
1310
fG•mA•fC•mC•fG•mC•fC•mA•fA•mG•fG•mA•fU•mG•fA


132
1311
fA•mC•fC•mG•fC•mC•fA•mA•fG•mG•fA•mU•fG•mC•fA


133
1312
fC•mC•fG•mC•fC•mA•fA•mG•fG•mA•fU•mG•fC•mA•fA


134
1313
fC•mG•fC•mC•fA•mA•fG•mG•fA•mU•fG•mC•fA•mC•fA


135
1314
fG•mC•fC•mA•fA•mG•fG•mA•fU•mG•fC•mA•fC•mU•fA


136
1315
fC•mC•fA•mA•fG•mG•fA•mU•fG•mC•fA•mC•fU•mG•fA


137
1316
fC•mA•fA•mG•fG•mA•fU•mG•fC•mA•fC•mU•fG•mA•fA


138
1317
fA•mA•fG•mG•fA•mU•fG•mC•fA•mC•fU•mG•fA•mG•fA


139
1318
fA•mG•fG•mA•fU•mG•fC•mA•fC•mU•fG•mA•fG•mC•fA


140
1319
fG•mG•fA•mU•fG•mC•fA•mC•fU•mG•fA•mG•fC•mA•fA


141
1320
fG•mA•fU•mG•fC•mA•fC•mU•fG•mA•fG•mC•fA•mG•fA


142
1321
fA•mU•fG•mC•fA•mC•fU•mG•fA•mG•fC•mA•fG•mC•fA


143
1322
fU•mG•fC•mA•fC•mU•fG•mA•fG•mC•fA•mG•fC•mG•fA


144
1323
fG•mC•fA•mC•fU•mG•fA•mG•fC•mA•fG•mC•fG•mU•fA


145
1324
fC•mA•fC•mU•fG•mA•fG•mC•fA•mG•fC•mG•fU•mG•fA


146
1325
fA•mC•fU•mG•fA•mG•fC•mA•fG•mC•fG•mU•fG•mC•fA


147
1326
fC•mU•fG•mA•fG•mC•fA•mG•fC•mG•fU•mG•fC•mA•fA


148
1327
fU•mG•fA•mG•fC•mA•fG•mC•fG•mU•fG•mC•fA•mG•fA


149
1328
fG•mA•fG•mC•fA•mG•fC•mG•fU•mG•fC•mA•fG•mG•fA


150
1329
fA•mG•fC•mA•fG•mC•fG•mU•fG•mC•fA•mG•fG•mA•fA


151
1330
fC•mA•fG•mC•fG•mU•fG•mC•fA•mG•fG•mA•fG•mU•fA


152
1331
fC•mG•fU•mG•fC•mA•fG•mG•fA•mG•fU•mC•fC•mC•fA


153
1332
fG•mU•fG•mC•fA•mG•fG•mA•fG•mU•fC•mC•fC•mA•fA


154
1333
fU•mG•fC•mA•fG•mG•fA•mG•fU•mC•fC•mC•fA•mG•fA


155
1334
fG•mC•fA•mG•fG•mA•fG•mU•fC•mC•fC•mA•fG•mG•fA


156
1335
fC•mA•fG•mG•fA•mG•fU•mC•fC•mC•fA•mG•fG•mU•fA


157
1336
fA•mG•fG•mA•fG•mU•fC•mC•fC•mA•fG•mG•fU•mG•fA


158
1337
fG•mG•fA•mG•fU•mC•fC•mC•fA•mG•fG•mU•fG•mG•fA


159
1338
fG•mU•fC•mC•fC•mA•fG•mG•fU•mG•fG•mC•fC•mC•fA


160
1339
fU•mC•fC•mC•fA•mG•fG•mU•fG•mG•fC•mC•fC•mA•fA


161
1340
fC•mA•fG•mG•fU•mG•fG•mC•fC•mC•fA•mG•fC•mA•fA


162
1341
fA•mG•fG•mU•fG•mG•fC•mC•fC•mA•fG•mC•fA•mG•fA


163
1342
fU•mG•fG•mC•fC•mC•fA•mG•fC•mA•fG•mG•fC•mC•fA


164
1343
fC•mC•fC•mA•fG•mC•fA•mG•fG•mC•fC•mA•fG•mG•fA


165
1344
fU•mG•fG•mG•fU•mG•fA•mC•fC•mG•fA•mU•fG•mG•fA


166
1345
fG•mG•fG•mU•fG•mA•fC•mC•fG•mA•fU•mG•fG•mC•fA


167
1346
fG•mG•fU•mG•fA•mC•fC•mG•fA•mU•fG•mG•fC•mU•fA


168
1347
fG•mU•fG•mA•fC•mC•fG•mA•fU•mG•fG•mC•fU•mU•fA


169
1348
fU•mG•fA•mC•fC•mG•fA•mU•fG•mG•fC•mU•fU•mC•fA


170
1349
fG•mA•fC•mC•fG•mA•fU•mG•fG•mC•fU•mU•fC•mA•fA


171
1350
fA•mC•fC•mG•fA•mU•fG•mG•fC•mU•fU•mC•fA•mG•fA


172
1351
fC•mC•fG•mA•fU•mG•fG•mC•fU•mU•fC•mA•fG•mU•fA


173
1352
fC•mG•fA•mU•fG•mG•fC•mU•fU•mC•fA•mG•fU•mU•fA


174
1353
fG•mA•fU•mG•fG•mC•fU•mU•fC•mA•fG•mU•fU•mC•fA


175
1354
fA•mU•fG•mG•fC•mU•fU•mC•fA•mG•fU•mU•fC•mC•fA


176
1355
fU•mG•fG•mC•fU•mU•fC•mA•fG•mU•fU•mC•fC•mC•fA


177
1356
fG•mG•fC•mU•fU•mC•fA•mG•fU•mU•fC•mC•fC•mU•fA


178
1357
fG•mC•fU•mU•fC•mA•fG•mU•fU•mC•fC•mC•fU•mG•fA


179
1358
fC•mU•fU•mC•fA•mG•fU•mU•fC•mC•fC•mU•fG•mA•fA


180
1359
fU•mU•fC•mA•fG•mU•fU•mC•fC•mC•fU•mG•fA•mA•fA


181
1360
fU•mC•fA•mG•fU•mU•fC•mC•fC•mU•fG•mA•fA•mA•fA


182
1361
fC•mA•fG•mU•fU•mC•fC•mC•fU•mG•fA•mA•fA•mG•fA


183
1362
fA•mG•fU•mU•fC•mC•fC•mU•fG•mA•fA•mA•fG•mA•fA


184
1363
fG•mU•fU•mC•fC•mC•fU•mG•fA•mA•fA•mG•fA•mC•fA


185
1364
fU•mU•fC•mC•fC•mU•fG•mA•fA•mA•fG•mA•fC•mU•fA


186
1365
fU•mC•fC•mC•fU•mG•fA•mA•fA•mG•fA•mC•fU•mA•fA


187
1366
fC•mC•fC•mU•fG•mA•fA•mA•fG•mA•fC•mU•fA•mC•fA


188
1367
fC•mC•fU•mG•fA•mA•fA•mG•fA•mC•fU•mA•fC•mU•fA


189
1368
fC•mU•fG•mA•fA•mA•fG•mA•fC•mU•fA•mC•fU•mG•fA


190
1369
fU•mG•fA•mA•fA•mG•fA•mC•fU•mA•fC•mU•fG•mG•fA


191
1370
fG•mA•fA•mA•fG•mA•fC•mU•fA•mC•fU•mG•fG•mA•fA


192
1371
fA•mA•fA•mG•fA•mC•fU•mA•fC•mU•fG•mG•fA•mG•fA


193
1372
fA•mA•fG•mA•fC•mU•fA•mC•fU•mG•fG•mA•fG•mC•fA


194
1373
fA•mG•fA•mC•fU•mA•fC•mU•fG•mG•fA•mG•fC•mA•fA


195
1374
fG•mA•fC•mU•fA•mC•fU•mG•fG•mA•fG•mC•fA•mC•fA


196
1375
fA•mC•fU•mA•fC•mU•fG•mG•fA•mG•fC•mA•fC•mC•fA


197
1376
fC•mU•fA•mC•fU•mG•fG•mA•fG•mC•fA•mC•fC•mG•fA


198
1377
fU•mA•fC•mU•fG•mG•fA•mG•fC•mA•fC•mC•fG•mU•fA


199
1378
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200
1379
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201
1380
fU•mG•fG•mA•fG•mC•fA•mC•fC•mG•fU•mU•fA•mA•fA


202
1381
fG•mG•fA•mG•fC•mA•fC•mC•fG•mU•fU•mA•fA•mG•fA


203
1382
fG•mA•fG•mC•fA•mC•fC•mG•fU•mU•fA•mA•fG•mG•fA


204
1383
fA•mG•fC•mA•fC•mC•fG•mU•fU•mA•fA•mG•fG•mA•fA


205
1384
fG•mC•fA•mC•fC•mG•fU•mU•fA•mA•fG•mG•fA•mC•fA


206
1385
fC•mA•fC•mC•fG•mU•fU•mA•fA•mG•fG•mA•fC•mA•fA


207
1386
fA•mC•fC•mG•fU•mU•fA•mA•fG•mG•fA•mC•fA•mA•fA


208
1387
fC•mC•fG•mU•fU•mA•fA•mG•fG•mA•fC•mA•fA•mG•fA


209
1388
fC•mG•fU•mU•fA•mA•fG•mG•fA•mC•fA•mA•fG•mU•fA


210
1389
fG•mU•fU•mA•fA•mG•fG•mA•fC•mA•fA•mG•fU•mU•fA


211
1390
fU•mU•fA•mA•fG•mG•fA•mC•fA•mA•fG•mU•fU•mC•fA


212
1391
fU•mA•fA•mG•fG•mA•fC•mA•fA•mG•fU•mU•fC•mU•fA


213
1392
fA•mA•fG•mG•fA•mC•fA•mA•fG•mU•fU•mC•fU•mC•fA


214
1393
fA•mG•fG•mA•fC•mA•fA•mG•fU•mU•fC•mU•fC•mU•fA


215
1394
fG•mG•fA•mC•fA•mA•fG•mU•fU•mC•fU•mC•fU•mG•fA


216
1395
fG•mA•fC•mA•fA•mG•fU•mU•fC•mU•fC•mU•fG•mA•fA


217
1396
fA•mC•fA•mA•fG•mU•fU•mC•fU•mC•fU•mG•fA•mG•fA


218
1397
fC•mA•fA•mG•fU•mU•fC•mU•fC•mU•fG•mA•fG•mU•fA


219
1398
fA•mA•fG•mU•fU•mC•fU•mC•fU•mG•fA•mG•fU•mU•fA


220
1399
fA•mG•fU•mU•fC•mU•fC•mU•fG•mA•fG•mU•fU•mC•fA


221
1400
fU•mU•fC•mU•fC•mU•fG•mA•fG•mU•fU•mC•fU•mG•fA


222
1401
fU•mC•fU•mC•fU•mG•fA•mG•fU•mU•fC•mU•fG•mG•fA


223
1402
fC•mU•fC•mU•fG•mA•fG•mU•fU•mC•fU•mG•fG•mG•fA


224
1403
fU•mC•fU•mG•fA•mG•fU•mU•fC•mU•fG•mG•fG•mA•fA


225
1404
fC•mU•fG•mA•fG•mU•fU•mC•fU•mG•fG•mG•fA•mU•fA


226
1405
fU•mG•fA•mG•fU•mU•fC•mU•fG•mG•fG•mA•fU•mU•fA


227
1406
fG•mA•fG•mU•fU•mC•fU•mG•fG•mG•fA•mU•fU•mU•fA


228
1407
fA•mG•fU•mU•fC•mU•fG•mG•fG•mA•fU•mU•fU•mG•fA


229
1408
fG•mU•fU•mC•fU•mG•fG•mG•fA•mU•fU•mU•fG•mG•fA


230
1409
fU•mU•fC•mU•fG•mG•fG•mA•fU•mU•fU•mG•fG•mA•fA


231
1410
fU•mC•fU•mG•fG•mG•fA•mU•fU•mU•fG•mG•fA•mC•fA


232
1411
fC•mU•fG•mG•fG•mA•fU•mU•fU•mG•fG•mA•fC•mC•fA


233
1412
fU•mG•fG•mG•fA•mU•fU•mU•fG•mG•fA•mC•fC•mC•fA


234
1413
fG•mG•fG•mA•fU•mU•fU•mG•fG•mA•fC•mC•fC•mU•fA


235
1414
fG•mG•fA•mU•fU•mU•fG•mG•fA•mC•fC•mC•fU•mG•fA


236
1415
fG•mA•fU•mU•fU•mG•fG•mA•fC•mC•fC•mU•fG•mA•fA


237
1416
fU•mG•fG•mA•fC•mC•fC•mU•fG•mA•fG•mG•fU•mC•fA


238
1417
fG•mG•fA•mC•fC•mC•fU•mG•fA•mG•fG•mU•fC•mA•fA


239
1418
fG•mA•fC•mC•fC•mU•fG•mA•fG•mG•fU•mC•fA•mG•fA


240
1419
fA•mC•fC•mC•fU•mG•fA•mG•fG•mU•fC•mA•fG•mA•fA


241
1420
fC•mC•fC•mU•fG•mA•fG•mG•fU•mC•fA•mG•fA•mC•fA


242
1421
fC•mC•fU•mG•fA•mG•fG•mU•fC•mA•fG•mA•fC•mC•fA


243
1422
fC•mU•fG•mA•fG•mG•fU•mC•fA•mG•fA•mC•fC•mA•fA


244
1423
fU•mG•fA•mG•fG•mU•fC•mA•fG•mA•fC•mC•fA•mA•fA


245
1424
fG•mA•fG•mG•fU•mC•fA•mG•fA•mC•fC•mA•fA•mC•fA


246
1425
fA•mG•fG•mU•fC•mA•fG•mA•fC•mC•fA•mA•fC•mU•fA


247
1426
fG•mG•fU•mC•fA•mG•fA•mC•fC•mA•fA•mC•fU•mU•fA


248
1427
fG•mU•fC•mA•fG•mA•fC•mC•fA•mA•fC•mU•fU•mC•fA


249
1428
fU•mC•fA•mG•fA•mC•fC•mA•fA•mC•fU•mU•fC•mA•fA


250
1429
fC•mA•fG•mA•fC•mC•fA•mA•fC•mU•fU•mC•fA•mG•fA


251
1430
fG•mA•fC•mC•fA•mA•fC•mU•fU•mC•fA•mG•fC•mC•fA


252
1431
fA•mC•fC•mA•fA•mC•fU•mU•fC•mA•fG•mC•fC•mG•fA


253
1432
fC•mC•fA•mA•fC•mU•fU•mC•fA•mG•fC•mC•fG•mU•fA


254
1433
fC•mA•fA•mC•fU•mU•fC•mA•fG•mC•fC•mG•fU•mG•fA


255
1434
fA•mA•fC•mU•fU•mC•fA•mG•fC•mC•fG•mU•fG•mG•fA


256
1435
fA•mC•fU•mU•fC•mA•fG•mC•fC•mG•fU•mG•fG•mC•fA


257
1436
fU•mU•fC•mA•fG•mC•fC•mG•fU•mG•fG•mC•fU•mG•fA


258
1437
fU•mC•fA•mG•fC•mC•fG•mU•fG•mG•fC•mU•fG•mC•fA


259
1438
fC•mA•fG•mC•fC•mG•fU•mG•fG•mC•fU•mG•fC•mC•fA


260
1439
fA•mG•fC•mC•fG•mU•fG•mG•fC•mU•fG•mC•fC•mU•fA


261
1440
fG•mC•fC•mG•fU•mG•fG•mC•fU•mG•fC•mC•fU•mG•fA


262
1441
fG•mU•fG•mG•fC•mU•fG•mC•fC•mU•fG•mA•fG•mA•fA


263
1442
fU•mG•fG•mC•fU•mG•fC•mC•fU•mG•fA•mG•fA•mC•fA


264
1443
fG•mG•fC•mU•fG•mC•fC•mU•fG•mA•fG•mA•fC•mC•fA


265
1444
fG•mC•fU•mG•fC•mC•fU•mG•fA•mG•fA•mC•fC•mU•fA


266
1445
fU•mG•fC•mC•fU•mG•fA•mG•fA•mC•fC•mU•fC•mA•fA


267
1446
fG•mC•fC•mU•fG•mA•fG•mA•fC•mC•fU•mC•fA•mA•fA


268
1447
fC•mC•fU•mG•fA•mG•fA•mC•fC•mU•fC•mA•fA•mU•fA


269
1448
fC•mU•fG•mA•fG•mA•fC•mC•fU•mC•fA•mA•fU•mA•fA


270
1449
fU•mG•fA•mG•fA•mC•fC•mU•fC•mA•fA•mU•fA•mC•fA


271
1450
fG•mA•fG•mA•fC•mC•fU•mC•fA•mA•fU•mA•fC•mC•fA


272
1451
fA•mG•fA•mC•fC•mU•fC•mA•fA•mU•fA•mC•fC•mC•fA


273
1452
fA•mG•fU•mC•fC•mA•fC•mC•fU•mG•fC•mC•fU•mA•fA


274
1453
fG•mU•fC•mC•fA•mC•fC•mU•fG•mC•fC•mU•fA•mU•fA


275
1454
fU•mC•fC•mA•fC•mC•fU•mG•fC•mC•fU•mA•fU•mC•fA


276
1455
fC•mC•fA•mC•fC•mU•fG•mC•fC•mU•fA•mU•fC•mC•fA


277
1456
fC•mA•fC•mC•fU•mG•fC•mC•fU•mA•fU•mC•fC•mA•fA


278
1457
fA•mC•fC•mU•fG•mC•fC•mU•fA•mU•fC•mC•fA•mU•fA


279
1458
fC•mC•fU•mG•fC•mC•fU•mA•fU•mC•fC•mA•fU•mC•fA


280
1459
fC•mU•fG•mC•fC•mU•fA•mU•fC•mC•fA•mU•fC•mC•fA


281
1460
fU•mG•fC•mC•fU•mA•fU•mC•fC•mA•fU•mC•fC•mU•fA


282
1461
fG•mC•fC•mU•fA•mU•fC•mC•fA•mU•fC•mC•fU•mG•fA


283
1462
fC•mC•fU•mA•fU•mC•fC•mA•fU•mC•fC•mU•fG•mC•fA


284
1463
fC•mU•fA•mU•fC•mC•fA•mU•fC•mC•fU•mG•fC•mG•fA


285
1464
fU•mA•fU•mC•fC•mA•fU•mC•fC•mU•fG•mC•fG•mA•fA


286
1465
fA•mU•fC•mC•fA•mU•fC•mC•fU•mG•fC•mG•fA•mG•fA


287
1466
fU•mC•fC•mA•fU•mC•fC•mU•fG•mC•fG•mA•fG•mC•fA


288
1467
fC•mC•fA•mU•fC•mC•fU•mG•fC•mG•fA•mG•fC•mU•fA


289
1468
fC•mA•fU•mC•fC•mU•fG•mC•fG•mA•fG•mC•fU•mC•fA


290
1469
fA•mU•fC•mC•fU•mG•fC•mG•fA•mG•fC•mU•fC•mC•fA


291
1470
fU•mC•fC•mU•fG•mC•fG•mA•fG•mC•fU•mC•fC•mU•fA


292
1471
fC•mC•fU•mG•fC•mG•fA•mG•fC•mU•fC•mC•fU•mU•fA


293
1472
fC•mU•fG•mC•fG•mA•fG•mC•fU•mC•fC•mU•fU•mG•fA


294
1473
fU•mG•fC•mG•fA•mG•fC•mU•fC•mC•fU•mU•fG•mG•fA


295
1474
fG•mC•fG•mA•fG•mC•fU•mC•fC•mU•fU•mG•fG•mG•fA


296
1475
fC•mG•fA•mG•fC•mU•fC•mC•fU•mU•fG•mG•fG•mU•fA


297
1476
fG•mA•fG•mC•fU•mC•fC•mU•fU•mG•fG•mG•fU•mC•fA


298
1477
fA•mG•fC•mU•fC•mC•fU•mU•fG•mG•fG•mU•fC•mC•fA


299
1478
fG•mC•fU•mC•fC•mU•fU•mG•fG•mG•fU•mC•fC•mU•fA


300
1479
fC•mU•fC•mC•fU•mU•fG•mG•fG•mU•fC•mC•fU•mG•fA


301
1480
fU•mC•fC•mU•fU•mG•fG•mG•fU•mC•fC•mU•fG•mC•fA


302
1481
fC•mC•fU•mU•fG•mG•fG•mU•fC•mC•fU•mG•fC•mA•fA


303
1482
fC•mU•fU•mG•fG•mG•fU•mC•fC•mU•fG•mC•fA•mA•fA


304
1483
fU•mU•fG•mG•fG•mU•fC•mC•fU•mG•fC•mA•fA•mU•fA


305
1484
fU•mG•fG•mG•fU•mC•fC•mU•fG•mC•fA•mA•fU•mC•fA


306
1485
fG•mG•fG•mU•fC•mC•fU•mG•fC•mA•fA•mU•fC•mU•fA


307
1486
fG•mG•fU•mC•fC•mU•fG•mC•fA•mA•fU•mC•fU•mC•fA


308
1487
fG•mU•fC•mC•fU•mG•fC•mA•fA•mU•fC•mU•fC•mC•fA


309
1488
fU•mC•fC•mU•fG•mC•fA•mA•fU•mC•fU•mC•fC•mA•fA


310
1489
fC•mC•fU•mG•fC•mA•fA•mU•fC•mU•fC•mC•fA•mG•fA


311
1490
fC•mU•fG•mC•fA•mA•fU•mC•fU•mC•fC•mA•fG•mG•fA


312
1491
fU•mG•fC•mA•fA•mU•fC•mU•fC•mC•fA•mG•fG•mG•fA


313
1492
fG•mC•fA•mA•fU•mC•fU•mC•fC•mA•fG•mG•fG•mC•fA


314
1493
fC•mA•fA•mU•fC•mU•fC•mC•fA•mG•fG•mG•fC•mU•fA


315
1494
fA•mA•fU•mC•fU•mC•fC•mA•fG•mG•fG•mC•fU•mG•fA


316
1495
fA•mU•fC•mU•fC•mC•fA•mG•fG•mG•fC•mU•fG•mC•fA


317
1496
fU•mC•fU•mC•fC•mA•fG•mG•fG•mC•fU•mG•fC•mC•fA


318
1497
fC•mU•fC•mC•fA•mG•fG•mG•fC•mU•fG•mC•fC•mC•fA


319
1498
fG•mU•fA•mG•fG•mU•fU•mG•fC•mU•fU•mA•fA•mA•fA


320
1499
fU•mA•fG•mG•fU•mU•fG•mC•fU•mU•fA•mA•fA•mA•fA


321
1500
fA•mG•fG•mU•fU•mG•fC•mU•fU•mA•fA•mA•fA•mG•fA


322
1501
fG•mG•fU•mU•fG•mC•fU•mU•fA•mA•fA•mA•fG•mG•fA


323
1502
fG•mU•fU•mG•fC•mU•fU•mA•fA•mA•fA•mG•fG•mG•fA


324
1503
fU•mU•fG•mC•fU•mU•fA•mA•fA•mA•fG•mG•fG•mA•fA


325
1504
fU•mG•fC•mU•fU•mA•fA•mA•fA•mG•fG•mG•fA•mC•fA


326
1505
fU•mU•fA•mA•fA•mA•fG•mG•fG•mA•fC•mA•fG•mU•fA


327
1506
fU•mA•fA•mA•fA•mG•fG•mG•fA•mC•fA•mG•fU•mA•fA


328
1507
fA•mA•fA•mA•fG•mG•fG•mA•fC•mA•fG•mU•fA•mU•fA


329
1508
fA•mA•fA•mG•fG•mG•fA•mC•fA•mG•fU•mA•fU•mU•fA


330
1509
fA•mA•fG•mG•fG•mA•fC•mA•fG•mU•fA•mU•fU•mC•fA


331
1510
fA•mG•fG•mG•fA•mC•fA•mG•fU•mA•fU•mU•fC•mU•fA


332
1511
fG•mG•fG•mA•fC•mA•fG•mU•fA•mU•fU•mC•fU•mC•fA


333
1512
fG•mG•fA•mC•fA•mG•fU•mA•fU•mU•fC•mU•fC•mA•fA


334
1513
fG•mA•fC•mA•fG•mU•fA•mU•fU•mC•fU•mC•fA•mG•fA


335
1514
fA•mC•fA•mG•fU•mA•fU•mU•fC•mU•fC•mA•fG•mU•fA


336
1515
fC•mA•fG•mU•fA•mU•fU•mC•fU•mC•fA•mG•fU•mG•fA


337
1516
fA•mG•fU•mA•fU•mU•fC•mU•fC•mA•fG•mU•fG•mC•fA


338
1517
fG•mU•fA•mU•fU•mC•fU•mC•fA•mG•fU•mG•fC•mU•fA


339
1518
fU•mA•fU•mU•fC•mU•fC•mA•fG•mU•fG•mC•fU•mC•fA


340
1519
fA•mU•fU•mC•fU•mC•fA•mG•fU•mG•fC•mU•fC•mU•fA


341
1520
fU•mU•fC•mU•fC•mA•fG•mU•fG•mC•fU•mC•fU•mC•fA


342
1521
fU•mC•fU•mC•fA•mG•fU•mG•fC•mU•fC•mU•fC•mC•fA


343
1522
fC•mU•fC•mA•fG•mU•fG•mC•fU•mC•fU•mC•fC•mU•fA


344
1523
fU•mC•fA•mG•fU•mG•fC•mU•fC•mU•fC•mC•fU•mA•fA


345
1524
fC•mA•fG•mU•fG•mC•fU•mC•fU•mC•fC•mU•fA•mC•fA


346
1525
fA•mG•fU•mG•fC•mU•fC•mU•fC•mC•fU•mA•fC•mC•fA


347
1526
fG•mU•fG•mC•fU•mC•fU•mC•fC•mU•fA•mC•fC•mC•fA


348
1527
fC•mC•fU•mC•fA•mU•fG•mC•fC•mU•fG•mG•fC•mC•fA


349
1528
fC•mU•fC•mA•fU•mG•fC•mC•fU•mG•fG•mC•fC•mC•fA


350
1529
fC•mC•fA•mG•fG•mC•fA•mU•fG•mC•fU•mG•fG•mC•fA


351
1530
fC•mA•fG•mG•fC•mA•fU•mG•fC•mU•fG•mG•fC•mC•fA


352
1531
fA•mG•fG•mC•fA•mU•fG•mC•fU•mG•fG•mC•fC•mU•fA


353
1532
fG•mG•fC•mA•fU•mG•fC•mU•fG•mG•fC•mC•fU•mC•fA


354
1533
fG•mC•fA•mU•fG•mC•fU•mG•fG•mC•fC•mU•fC•mC•fA


355
1534
fC•mA•fU•mG•fC•mU•fG•mG•fC•mC•fU•mC•fC•mC•fA


356
1535
fA•mU•fG•mC•fU•mG•fG•mC•fC•mU•fC•mC•fC•mA•fA


357
1536
fG•mC•fU•mG•fG•mC•fC•mU•fC•mC•fC•mA•fA•mU•fA


358
1537
fC•mU•fG•mG•fC•mC•fU•mC•fC•mC•fA•mA•fU•mA•fA


359
1538
fU•mG•fG•mC•fC•mU•fC•mC•fC•mA•fA•mU•fA•mA•fA


360
1539
fG•mG•fC•mC•fU•mC•fC•mC•fA•mA•fU•mA•fA•mA•fA


361
1540
fG•mC•fC•mU•fC•mC•fC•mA•fA•mU•fA•mA•fA•mG•fA


362
1541
fC•mC•fU•mC•fC•mC•fA•mA•fU•mA•fA•mA•fG•mC•fA


363
1542
fC•mU•fC•mC•fC•mA•fA•mU•fA•mA•fA•mG•fC•mU•fA


364
1543
fU•mC•fC•mC•fA•mA•fU•mA•fA•mA•fG•mC•fU•mG•fA


365
1544
fC•mC•fC•mA•fA•mU•fA•mA•fA•mG•fC•mU•fG•mG•fA


366
1545
fC•mC•fA•mA•fU•mA•fA•mA•fG•mC•fU•mG•fG•mA•fA


367
1546
fC•mA•fA•mU•fA•mA•fA•mG•fC•mU•fG•mG•fA•mC•fA


368
1547
fA•mU•fA•mA•fA•mG•fC•mU•fG•mG•fA•mC•fA•mA•fA


369
1548
fU•mA•fA•mA•fG•mC•fU•mG•fG•mA•fC•mA•fA•mG•fA


370
1549
fA•mA•fA•mG•fC•mU•fG•mG•fA•mC•fA•mA•fG•mA•fA


371
1550
fA•mA•fG•mC•fU•mG•fG•mA•fC•mA•fA•mG•fA•mA•fA


372
1551
fA•mG•fC•mU•fG•mG•fA•mC•fA•mA•fG•mA•fA•mG•fA


373
1552
fG•mC•fU•mG•fG•mA•fC•mA•fA•mG•fA•mA•fG•mC•fA


374
1553
fG•mG•fA•mC•fA•mA•fG•mA•fA•mG•fC•mU•fG•mC•fA


375
1554
fA•mC•fA•mA•fG•mA•fA•mG•fC•mU•fG•mC•fU•mA•fA


376
1555
fC•mA•fA•mG•fA•mA•fG•mC•fU•mG•fC•mU•fA•mU•fA









Tables 2a to 2d below show nucleobase sequences and sugar-phosphate backbone modifications of antisense and sense strands of a further 15 exemplary constructs. For corresponding entries in the sequence listing, the following applies: entry number in Table 2a+376=entry number in the sequence listing; entry number in Table 2c+776=entry number in the sequence listing.







TABLE 2a







Nucleobase sequences of the antisense strands


of 15 further exemplary constructs










SEQ




ID



#
NO:
AS unmodified












1
377
UAACUCAGAGAACUUGUCC


 


2
378
UUGUCCUUAACGGUGCUCC


 


3
379
UAAUCCCAGAACUCAGAGA


 


4
380
UCCUUGGCGGUCUUGGUGG


 


5
381
UCUGAAGCCAUCGGUCACC


 


6
382
UCAGAGAACUUGUCCUUAA


 


7
383
UACUCAGAGAACUUGUCCU


 


8
384
UGAACUCAGAGAACUUGUC


 


9
385
UACUUGUCCUUAACGGUGC


 


10
386
UCUCAGAGAACUUGUCCUU


 


11
387
UUUGUCCUUAACGGUGCUC


 


12
388
UUCCUUGGCGGUCUUGGUG


 


13
389
UGCUCCAGUAGUCUUUCAG


 


14
390
UCAUCCUCGGCCUCUGAAG


 


15
391
UUGGUGGCGUGCUUCAUGU
















TABLE 2b







Nucleobase sequences and sugar-phosphate backbone modifications 


of the antisense strands of 15 further exemplary constructs:










SEQ




ID



#
NO:
Antisense strand modified





 1
1556
[mU][#][fA][#][mA][#][fC][mU][fC][mA][fG][mA][fG][mA][fA][mC][fU][#][mU][#][fG][#][mU][#][fC][#]rC


 2
1557
[mU][#][fU][#][mG][#][fU][mC][fC][mU][fU][mA][fA][mC][fG][mG][fU][#][mG][#][fC][#][mU][#][fC][#]rC


 3
1558
[mU][#][fA][#][mA][#][fU][mC][fC][mC][fA][mG][fA][mA][fC][mU][fC][#][mA][#][fG][#][mA][#][fG][#]rA


 4
1559
[mU][#][fC][#][mC][#][fU][mU][fG][mG][fC][mG][fG][mU][fC][mU][fU][#][mG][#][fG][#][mU][#][fG][#]rG


 5
1560
[mU][#][fC][#][mU][#][fG][mA][fA][mG][fC][mC][fA][mU][fC][mG][fG][#][mU][#][fC][#][mA][#][fC][#]rC


 6
1561
[mU][#][fC][#][mA][#][fG][mA][fG][mA][fA][mC][fU][mU][fG][mU][fC][#][mC][#][fU][#][mU][#][fA][#]rA


 7
1562
[mU][#][fA][#][mC][#][fU][mC][fA][mG][fA][mG][fA][mA][fC][mU][fU][#][mG][#][fU][#][mC][#][fC][#]rU


 8
1563
[mU][#][fG][#][mA][#][fA][mC][fU][mC][fA][mG][fA][mG][fA][mA][fC][#][mU][#][fU][#][mG][#][fU][#]rC


 9
1564
[mU][#][fA][#][mC][#][fU][mU][fG][mU][fC][mC][fU][mU][fA][mA][fC][#][mG][#][fG][#][mU][#][fG][#]rC


10
1565
[mU][#][fC][#][mU][#][fC][mA][fG][mA][fG][mA][fA][mC][fU][mU][fG][#][mU][#][fC][#][mC][#][fU][#]rU


11
1566
[mU][#][fU][#][mU][#][fG][mU][fC][mC][fU][mU][fA][mA][fC][mG][fG][#][mU][#][fG][#][mC][#][fU][#]rC


12
1567
[mU][#][fU][#][mC][#][fC][mU][fU][mG][fG][mC][fG][mG][fU][mC][fU][#][mU][#][fG][#][mG][#][fU][#]rG


13
1568
[mU][#][fG][#][mC][#][fU][mC][fC][mA][fG][mU][fA][mG][fU][mC][fU][#][mU][#][fU][#][mC][#][fA][#]rG


14
1569
[mU][#][fC][#][mA][#][fU][mC][fC][mU][fC][mG][fG][mC][fC][mU][fC][#][mU][#][fG][#][mA][#][fA][#]rG


15
1570
[mU][#][fU][#][mG][#][fG][mU][fG][mG][fC][mG][fU][mG][fC][mU][fU][#][mC][#][fA][#][mU][#][fG][#]rU
















TABLE 2c







Nucleobase sequences of the sense strands


of 15 further exemplary constructs












SEQ





ID




#
NO:
SS unmodified















1
777
AGUUCUCUGAGUUA



 



2
778
ACCGUUAAGGACAA



 



3
779
GAGUUCUGGGAUUA



 



4
780
AAGACCGCCAAGGA



 



5
781
CCGAUGGCUUCAGA



 



6
782
GACAAGUUCUCUGA



 



7
783
AAGUUCUCUGAGUA



 



8
784
GUUCUCUGAGUUCA



 



9
785
GUUAAGGACAAGUA



 



10
786
CAAGUUCUCUGAGA



 



11
787
CCGUUAAGGACAAA



 



12
788
AGACCGCCAAGGAA



 



13
789
AGACUACUGGAGCA



 



14
790
GAGGCCGAGGAUGA



 



15
791
AAGCACGCCACCAA

















TABLE 2d







Nucleobase sequences and sugar-phosphate backbone modifications 


of the sense strands of 15 further exemplary constructs:










SEQ




ID



#
NO:
Sense strand modified





 1
1571
[mA][#][fG][#][mU][fU][mC][fU][mC][fU][mG][fA][mG][fU][#][mU][#][fA][#][3 × 




GalNac]


 2
1572
[mA][#][fC][#][mC][fG][mU][fU][mA][fA][mG][fG][mA][fC][#][mA][#][fA][#][3 × 




GalNac]


 3
1573
[mG][#][fA][#][mG][fU][mU][fC][mU][fG][mG][fG][mA][fU][#][mU][#][fA][#][3 × 




GalNac]


 4
1574
[mA][#][fA][#][mG][fA][mC][fC][mG][fC][mC][fA][mA][fG][#][mG][#][fA][#][3 × 




GalNac]


 5
1575
[mC][#][fC][#][mG][fA][mU][fG][mG][fC][mU][fU][mC][fA][#][mG][#][fA][#][3 × 




GalNac]


 6
1576
[mG][#][fA][#][mC][fA][mA][fG][mU][fU][mC][fU][mC][fU][#][mG][#][fA][#][3 × 




GalNac]


 7
1577
[mA][#][fA][#][mG][fU][mU][fC][mU][fC][mU][fG][mA][fG][#][mU][#][fA][#][3 × 




GalNac]


 8
1578
[mG][#][fU][#][mU][fC][mU][fC][mU][fG][mA][fG][mU][fU][#][mC][#][fA][#][3 × 




GalNac]


 9
1579
[mG][#][fU][#][mU][fA][mA][fG][mG][fA][mC][fA][mA][fG][#][mU][#][fA][#][3 × 




GalNac]


10
1580
[mC][#][fA][#][mA][fG][mU][fU][mC][fU][mC][fU][mG][fA][#][mG][#][fA][#][3 × 




GalNac]


11
1581
[mC][#][fC][#][mG][fU][mU][fA][mA][fG][mG][fA][mC][fA][#][mA][#][fA][#][3 × 




GalNac]


12
1582
[mA][#][fG][#][mA][fC][mC][fG][mC][fC][mA][fA][mG][fG][#][mA][#][fA][#][3 × 




GalNac]


13
1583
[mA][#][fG][#][mA][fC][mU][fA][mC][fU][mG][fG][mA][fG][#][mC][#][fA][#][3 × 




GalNac]


14
1584
[mG][#][fA][#][mG][fG][mC][fC][mG][fA][mG][fG][mA][fU][#][mG][#][fA][#][3 × 




GalNac]


15
1585
[mA][#][fA][#][mG][fC][mA][fC][mG][fC][mC][fA][mC][fC][#][mA][#][fA][#][3 × 




GalNac]
















TABLE 3a







Nucleobase sequences of the strands


of 12 further exemplary constructs.










SEQ




ID



#
NO:
Strands unmodified





A277(15)
792
uuggauaggc agguggacuc accugccuau




ccaa


 


A28(15)
793
ucaacaagga guacccgggg guacuccuug




uuga


 


A277(14)
794
uuggauaggc agguggacua ccugccuauc




caa


 


A28(14)
795
ucaacaagga guacccgggg uacuccuugu




uga


 


A277(12-5)
796
uuggauaggc agguggacug ccuauccaa


 


A277(13-4)
797
uuggauaggc agguggacuu gccuauccaa


 


A28(14-4)
798
ucaacaagga guacccgggu acuccuuguu ga


 


A277(14)mF
799
uuggauaggc agguggacua ccugccuauc




caa


 


A28(14)mF
800
ucaacaagga guacccgggg uacuccuugu




uga


 


A277(12-5)mF
801
uuggauaggc agguggacug ccuauccaa


 


A277(13-4)mF
802
uuggauaggc agguggacuu gccuauccaa


 


A28(14-4)mF
803
ucaacaagga guacccgggu acuccuuguu ga





Tables 3a to 3b below show nucleobase sequences and sugar-phosphate backbone modifications of 12 further exemplary constructs.













TABLE 3b







Nucleobase sequences and sugar-phosphate backbone modifications 


of the strands of 12 further exemplary constructs:









#
SEQ ID NO:
Strands modified





A277(15)
1586
[mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][mG][fU][mG][#][fG]




[#][mA][#][fC][#][mU][#][fC][mA][fC][mC][fU][mG][fC][mC][fU][mA][fU][mC]




[mC][#][mA][#][mA][#][3 × GalNAc]


A28(15)
1587
[mU][#][fC][#][mA][fA][mC][fA][mA][fG][mG][fA][mG][fU][mA][fC][mC][#][fC][#]




[mG][#][fG][#][mG][#][fG][mG][fU][mA][fC][mU][fC][mC][fU][mU][fG][mU][mU]




[#][mG][#][mA][#][3 × GalNAc]


A277(14)
1588
[mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][mG][fU][#][mG][#]




[fG][#][mA][#][fC][#][mU][#][mA][fC][mC][fU][mG][fC][mC][fU][mA][fU][mC][mC]




[#][mA][#][mA][#][3 × GalNAc]


A28(14)
1589
[mU][#][fC][#][mA][fA][mC][fA][mA][fG][mG][fA][mG][fU][mA][fC][#][mC][#][fC]




[#][mG][#][fG][#][mG][#][mG][fU][mA][fC][mU][fC][mC][fU][mU][fG][mU][mU]




[#][mG][#][mA][#][3 × GalNAc]


A277(12-5)
1590
[mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][#][mG][#][fU][mG][#]




[fG][#][mA][#][fC][fU][mG][fC][mC][fU][mA][fU][mC][mC][#][mA][#][mA][#][3 × 




GalNAc]


A277(13-4)
1591
[mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][mG][#][fU][#][mG][#]




[fG][#][mA][#][fC][mU][fU][mG][fC][mC][fU][mA][fU][mC][mC][#][mA][#][mA]




[#][3 × GalNac]


A28(14-4)
1592
[mU][#][fC][#][mA][fA][mC][fA][mA][fG][mG][fA][mG][fU][mA][fC][#][mC][#][fC]




[#][mG][#][fG][#][mG][fU][mA][fC][mU][fC][mC][fU][mU][fG][mU][mU][#][mG]




[#][mA][#][3 × GalNAc]


A277(14)mF
1593
[mU][#][fU][#][mG][mG][mA][mU][mA][mG][mG][mC][mA][mG][mG][fU][#][mG]




[#][mG][#][mA][#][mC][#][mU][#][mA][fC][fC][fU][mG][mC][mC][mU][mA][mU]




[mC][mC][#][mA][#][mA][#][3 × GalNac]


A28(14)mF
1594
[mU][#][fC][#][mA][mA][mC][mA][mA][mG][mG][mA][mG][mU][mA][fC][#][mC]




[#][mC][#][mG][#][mG][#][mG][#][mG][fU][fA][fC][mU][mC][mC][mU][mU][mG]




[mU][mU][#][mG][#][mA][#][3 × GalNAc]


A277(12-
1595
[mU][#][fU][#][mG][mG][mA][mU][mA][mG][mG][mC][mA][mG][#][mG][#][fU]


5) mF

[mG][#][mG][#][fA][#][fC][fU][mG][mC][mC][mU][mA][mU][mC][mC][#][mA][#]




[mA][#][3 × GalNAc]


A277(13-
1596
[mU][#][fU][#][mG][mG][mA][mU][mA][mG][mG][mC][mA][mG][mG][#][fU][#]


4) mF

[mG][#][mG][#][mA][#][fC][fU][fU][mG][mC][mC][mU][mA][mU][mC][mC][#]




[mA][#][mA][#][3 × GalNAc]


A28(14-4)mF
1597
[mU][#][fC][#][mA][mA][mC][mA][mA][mG][mG][mA][mG][mU][mA][fC][#][mC]




[#][mC][#][mG][#][mG][#][mG][fU][fA][fC][mU][mC][mC][mU][mU][mG][mU]




[mU][#][mG][#][mA][#][3 × GalNAc]









It should also be noted that the scope of the compositions and methods described herein extends to sequences that correspond to those in the Tables above, and wherein the 5′ nucleoside of the antisense (guide) strand (first region as defined in the items herein) can include any nucleobase that can be present in an RNA molecule, in other words can be any of adenine (A), uracil (U), guanine (G) or cytosine (C). Additionally, the scope of the present compositions and methods extends to sequences that correspond to those in Table 1a or Table 1 b, and wherein the 3′ nucleoside of the sense (passenger) strand (second region as defined in the items herein) can include any nucleobase that can be present in an RNA molecule, in other words can be any of adenine (A), uracil (U), guanine (G) or cytosine (C), preferably however a nucleobase that is complementary to the 5′ nucleobase of the antisense (guide) strand (first region as defined in the items herein).
While the methods are shown and described as being a series of acts that are performed in a particular sequence, it is to be understood and appreciated that the methods are not limited by the order of the sequence. For example, some acts can occur in a different order than what is described herein. In addition, an act can occur concurrently with another act. Further, in some instances, not all acts may be required to implement a method described herein.
The order of the steps of the methods described herein is exemplary, but the steps may be carried out in any suitable order, or simultaneously where appropriate. Additionally, steps may be added or substituted in, or individual steps may be deleted from any of the methods without departing from the scope of the subject matter described herein. Aspects of any of the Examples described above may be combined with aspects of any of the other Examples described to form further Examples.
It will be understood that the above description of a preferred embodiment is given by way of example only and that various modifications may be made by those skilled in the art. What has been described above includes Examples of one or more embodiments. It is, of course, not possible to describe every conceivable modification and alteration of the above compounds, compositions or methods for purposes of describing the aforementioned aspects, but one of ordinary skill in the art can recognize that many further modifications and permutations of various aspects are possible. Accordingly, the described aspects are intended to embrace all such alterations, modifications, and variations that fall within the scope of the appended claims.
EXAMPLES
The following Examples illustrate certain embodiments of the present disclosure and are not limiting. Moreover, where specific embodiments are provided, the generic application of those specific embodiments is contemplated. For example, disclosure of an oligonucleotide having a particular motif or modification patterns provides reasonable support for additional oligonucleotides having the same or similar motif or modification patterns.
The syntheses of the RNAi constructs as disclosed herein have been carried out using synthesis methods known to the person skilled in the art, such as synthesis methods disclosed in https://en.wikipedia.org/wiki/Oligonucleotide_synthesis {retrieved on 16 Feb. 2022}, wherein the methods disclosed on this website are incorporated by reference herein in their entirety. The only difference to the synthesis method disclosed in this reference is that GalNac phosphoramidite immobilized on a support is used in the synthesis method during the first synthesis step.
Example 1
Materials and Methods
Cell Culture:
HepG2 (ATCC cat. 85011430) cells were maintained by biweekly passing in EMEM supplemented with 10% FBS, 20 mM L-glutamine, 10 mM HEPES pH 7.2, 1 mM sodium pyruvate, 1×MEM non-essential amino acids, and 1×Pen/Strep (EMEM complete).
APOC3 Target identification and duplex preparation:
Targets to APOC3 were identified by bioinformatic analysis on human APOC3 mRNA sequence as given in RefSeq sequence ID NM_000040, wherein inter alia it has been taken into consideration that constructs as described herein should target APOC3 mRNA irrespective of splice variants and isoforms. 376 targets were selected for synthesis as asymmetric duplexes (14 nucleotide sense strand, 19 nucleotide antisense strand). Compounds were dissolved to 50 uM in molecular biology grade water and annealed by heating at 95 C for 5 minutes followed by gradual cooling to room temperature.
APOC3-Primary Screen:
On the day of transfection, HepG2 cells were collected by trypsinization, counted, and seeded in 96 well tissue culture treated plates at 10,000 cells per well in 50 uL complete EMEM with 20% FBS. Cells were allowed to rest for 4 hours before transfection with 2 pmoles of each respective APOC3 duplex in triplicate via RNAiMax (ThermoFisher). In brief, 8 pmoles of each duplex were diluted in 100 uL OptiMEM and mixed gently with 0.8 uL of RNAiMax in 100 uL OptiMEM to make 200 uL total complex. 50 uL of each RNAiMax complexed duplex was added to each respective triplicate well of HepG2 cells for a final mixture of 20 nM duplex in a volume of 100 uL, 50/50 EMEM/OptiMEM at 10% FBS.
72 hours post transfection, cells were harvested and RNA isolated using the PureLink Pro 96 total RNA Purification Kit (ThermoFisher, 12173011A) according to the manufacturer protocol. Harvested RNA was assayed for APOC3 expression via Taqman qPCR using the Luna Universal Probe One-Step RT-qPCR Kit (NEB, E3006). Two separate qPCR assays were performed for each sample using two separate APOC3 Taqman probe sets multiplexed with a common GAPDH VIC probe (ThermoFisher, 4326317E). Thermocycling and data acquisition was performed with an Applied Biosystems QuantStudio 3 Real-Time PCR System. Based on the results of the primary screen, a subset of 77 oligomeric compounds was selected which exhibit at least 70% target knockdown when assessed with either probe. These 77 compounds are defined by above items 3 and 4.
APOC3-Secondary Screen:
Based on data from the primary screen, a yet narrower set of the best performing 30 APOC3 duplexes were tested in dose curves. As before, HepG2 cells were collected by trypsinization and seeded in 96 well tissue culture plates at 10,000 cells per well in 50 uL complete EMEM with 20% FBS and allowed to rest for 4 hours. Transfection complexes were formed by gently mixing 36 pmoles of each duplex in 180 uL OptiMEM with 2.16 uL RNAiMax in 180 uL OptiMEM to make 360 uL total complex. A two fold dilution series was then performed with basal OptiMEM. 50 uL of each dilution was added to respective triplicates of HepG2 cells to make a final dilution series of 50 nM down to 0.32 nM in a volume of 100 uL, 50/50 EMEM/OptiMEM at 10% FBS.
72 hours post transfection, cells were harvested and RNA isolated using the PureLink Pro 96 total RNA Purification Kit (ThermoFisher, 12173011A) according to the manufacturer protocol. Harvested RNA was assayed for APOC3 expression via Taqman qPCR using the Luna Universal Probe One-Step RT-qPCR Kit (NEB, E3006). A single qPCR assay was performed for each sample using APOC3 Taqman probe set multiplexed with a common GAPDH VIC probe (ThermoFisher, 4326317E). Thermocycling and data acquisition was performed with an Applied Biosystems QuantStudio 3 Real-Time PCR System.
Example 2
Results
Table 4 below shows IC50 values (in nM) for the 30 constructs selected in accordance with the Examples.












Sequence ID
% k/d at the highest conc.
IC50

















AP277
93.44
3.29


AP337
93.10
4.10


AP028
90.64
4.53


AP343
93.10
4.70


AP369
90.15
4.86


AP366
95.63
5.56


AP274
89.43
5.89


AP367
88.85
5.99


AP336
92.76
6.13


AP332
90.23
6.35


AP293
84.99
6.44


AP373
89.76
6.46


AP280
78.85
6.71


AP221
92.66
6.84


AP334
90.35
6.85


AP286
83.77
6.89


AP149
90.36
7.77


AP193
91.30
7.83


AP328
87.02
7.85


AP175
94.58
8.28


AP262
84.65
8.72


AP254
90.79
9.11


AP185
88.83
9.20


AP328
88.99
9.44


AP271
78.49
9.49


AP137
86.09
9.79


AP225
81.11
10.74


AP167
84.77
11.13


AP297
84.99
13.28


AP191
84.23
14.27









The IC50 data in the single- to double-digit nanomolar range demonstrate outstanding performance of numerous constructs as described herein.
Example 3
Materials and Methods
Cell Culture:
Human primary hepatocytes (5 donor pooled—Sekisui XenoTech, HPCH05+) were thawed immediately prior to experimentation and cultured in 1×complete Williams medium (Gibco, A1217601) supplemented with Hepatocytes plating supplement pack (Gibco, CM3000). FBS concentration was modified from manufacture recipe to a final 2.5% (as opposed to 5%) for compound stability. 1×Complete WEM: 2.5% FBS, 1 μM Dexamethasone, Pen/Strep (100 U/mL/100 μg/mL), 4 μg/ml Human Insulin, 2 mM GlutaMAX, 15 mM HEPES, pH 7.4).
Hepatocytes were plated on Collagen I (rat tail) coated 96 well tissue culture plates (Gibco, A1142803).
APOC3 Compound Preparation:
Compounds were dissolved to 10 mg/mL in PBS and annealed by heating at 95 C for 5 minutes followed by rapid cooling on ice.
APOC3 Compound Transfections:
On the day of transfection, primary human hepatocytes were thawed in 45 mL of human OptiThaw (Sekisui Xenotech, K8000) and centrifuged down at 200 g for 5 minutes. Cells were resuspended in 2×complete WEM and counted. Cell were then plated in 50 uL of 2×complete WEM at 25,000 cells per well on 96 well type 1 rat tail Collagen plates and allowed to rest and attach for four hours before transfection.
Compounds were diluted further to 2 uM in basal WEM. A seven step, five fold dilution series was prepared in basal WEM from 2 uM to 0.000128 uM. 50 uL of each dilution was added to respective triplicates of the plated hepatocytes for a final dilution series of 1 uM down to 0.000064 uM in a volume of 100 uL 1×complete WEM.
72 hours post transfection, cells were harvested and RNA isolated using the PureLink Pro 96 total RNA Purification Kit (ThermoFisher, 12173011A) according to the manufacturer protocol. Harvested RNA was assayed for APOC3 expression via Taqman qPCR using the Luna Universal Probe One-Step RT-qPCR Kit (NEB, E3006). A single qPCR assay was performed for each sample using an APOC3 Taqman probe set (Hs00906501_g1-FAM) multiplexed with a common GAPDH VIC probe
(ThermoFisher, 4326317E). Thermocycling and data acquisition was performed with an Applied Biosystems QuantStudio 3/5 Real-Time PCR System.







TABLE 5





Constructs used as positive control
















A277(15)dup
5′



[mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][mG][fU][mG][#][fG][#][mA][#][fC][#][rU]



(SEQ ID NO: 1598)



5′ [fC][#][mA][#][fC][mC][fU][mG][fC][mC][fU][mA][fU][mC][mC][#][mA][#][mA][#][3 × GalNAc]



(SEQ ID NO: 1599)


A28(15)dup
5′



[mU][#][fC][#][mA][fA][mC][fA][mA][fG][mG][fA][mG][fU][mA][fC][mC][#][fC][#][mG][#][fG][#][rG]



(SEQ ID NO: 1600)



5′ [fG][#][mG][#][fU][mA][fC][mU][fC][mC][fU][mU][fG][mU][mU][#][mG][#][mA][#][3 × GalNAc]



(SEQ ID NO: 1601)


P29-A28
5′ [mU][#][fG][#][mC][fA][mA][fA][mA][fC][mA][fG][mG][fU][mC][fU][#][mA][#][fG][#][mA][#]



[fA][#][rA][mG][#][fU][#][mA][fC][mU][fC][mC][fU][mU][fG][mU][mU][#][mG][#][mA][#][3 × galNAc]



(SEQ ID NO: 1602)



5′ [mU][#][fC][#][mA][fA][mC][fA][mA][fG][mG][fA][mG][fU][mA][fC][#][mC][#][fC][#][mG][#][fG][#]



[rG][mA][#][fG][#][mA][fC][mC][fU][mG][fU][mU][fU][mU][mG][#][mC][#][mA][#][3 × GalNAc]



(SEQ ID NO: 1603)


P29-A277
5′ [mU][#][fG][#][mC][fA][mA][fA][mA][fC][mA][fG][mG][fU][mC][fU][#][mA][#][fG][#][mA][#][fA][#]



[rA][mA][#][fC][#][mC][fU][mG][fC][mC][fU][mA][fU][mC][mC][#][mA][#][mA][#][3 × GalNac] (SEQ



ID NO: 1604)



5′ [mU][#][fU][#][mG][fG][mA][fU][mA][fG][mG][fC][mA][fG][mG][fU][#][mG][#][fG][#][mA][#]



[fC][#][rU][mA][#][fG][#][mA][fC][mC][fU][mG][fU][mU][fU][mU][mG][#][mC][#][mA][#][3 × 



GalNAc] (SEQ ID NO: 1605)


TMPRSS6
5′ vP[mA][fA][mC][fC][mA][fG][mA][fA][mG][fA][mA][fG][mC][fA][mG][fG][mU][fG][iN][fC][mU]



[fG][fC][fU][mU][fC][mU][fU][mC][fU][mG][fG][mU][fU]#[3 × GalNAc] (SEQ ID NO: 1606)





Note:


vP = vinyl-phosphonate;


iN = inverted with 2′OH






Results

As can be seen from FIG. 1a, several variations of both A28 and A277 structures demonstrated excellent activities.
As can be seen from FIG. 1b, all molecules produced excellent activities.
Example 4
Study Protocol
The following study protocol for the study entitled “mxRNA Leads for Candidate Screening Study in Male human liver-uPA-SCID mice, non-GLP” has been drafted before the animal experiments and studies have been completed and therefore uses the future tense. However, as said study has already been completely carried out, each usage of “future tense” should be considered as the “past tense” in the following description of the study protocol.
Study Objective(s)
The objective of this non-GLP study was to evaluate the dose and duration response effect of two selected mxRNA leads for candidate GaINAc-siRNA constructs targeting APOC3 using the human liver-uPA-SCID mice models. The compounds were administered subcutaneously and the mice survived for 14-days and 42-days.
Prior to necropsy, plasma and serum were collected. At necropsy, 3 liver biopsies (2 mm) per animal were preserved in separate vials in RNAlater, flash frozen, and stored at −80° C. Three more liver biopsies (2 mm) were taken, flash frozen in the same vial, and stored at −80° C.
Regulatory Compliance
This non-GLP study will not be conducted in accordance with the Food and Drug Administration's Good Laboratory Practice (GLP) regulations (21 CFR Part 58).
Animal Welfare Compliance
The procedures described and performed below will be conducted in accordance with the Guide for the Care and Use of Laboratory Animals, USDA APHIS, Animal Welfare Act and/or in accordance with the Standard Operating Procedures.
This protocol has been reviewed and approved by the Test Facility IACUC Committee.
Study Schedule


    
    
        Acclimatization/Quarantine End Date: ≥5 days
        Baseline Procedure Date: No baseline procedures Procedure Start Day 0 Date: Tentative: December Waiting on test material.
        Necropsy Start: On Day 14- and 42-days post treatment.
        In-Life Study Completion: 6 weeks post treatment
        Preliminary Report: None required by Sponsor, Data only
        Final Report Issued: None required

Test System Information

        Animal Test
        Common Name: Mouse
        Breed/Class: Rodent—human liver-uPA-SCID mouse
        Number of Animals (by gender): 36 Male, all naïve
        Age Range: 14-19 weeks
        Weight Range: Approx. 20 grams
    
    


The mice used in this study were human liver-uPA-SCID mice. About 80% of the hepatocytes of each mouse have been replaced by human hepatocytes. The skilled person is aware of ways of producing such mice; wherein at least some of these ways are shown and referenced in P. Meuleman and G. Leroux-Roels in Antiviral Res. 2008 December; 80(3):231-8 which is incorporated herein by reference in its entirety.

    
    
        Acclimation Period:
        Duration:
    
    


All animals will be acclimated for a minimum period of five (5) days prior to release by the Attending veterinarian, at which time the overall health of the animals will be evaluated. Animals which are not released from acclimation will be treated accordingly and further evaluation will be performed prior to release. All records from the acclimation period will remain in the study file.
Animal Identification Method and Location:
Animals will be assigned sequential numbers. The animals will be ear notched to permanently identify each animal. This method involves punching holes or notches in the ear pinna while anesthetized.
Alternatively, the animals may have a tattoo placed on their tail. A cage card will also be affixed to each animal cage denoting the animal number, gender, vendor, strain, study director, and study number
Study Design
Design Details
This study will have one type of mice, 36 human liver-uPA-SCID mice. Animals will be grouped by treatment type, dosage, and survival period. Each animal will be treated by subcutaneous injection of test material. Groups 1A and 1B will have four animals receive a control dose of PBS. Groups 2A, 2B, 2C, 3A, 3B, and 3C will receive one dose (10 or 30 mg/kg) with four animals for each dose amount. All animals will be kept alive for 14 or 42 days. See study Table 6 below for details.







TABLE 6







Study Table













Number







of







human







liver-uPA-







SCID
Treatment






mice
Subcutaneous Injection
Survival

Pre-Euthanasia and


Group
animals
Day 0
Days
Blood
Necropsy















1A
4
Control (PBS)
14
Plasma and
Pre-Euthanasia:


1B
4
Control (PBS)
42
serum will
Plasma and serum






be
collection.


2A
4
A28 mxRNA (10 mg/kg)
14
collected
Necropsy:


2B
4
A28 mxRNA (30 mg/kg)
14
for all
2 mm biopsy of left,


2C
4
A28 mxRNA (10 mg/kg)
42
animals on
middle and right liver


3A
4
A277 mxRNA (10 mg/kg)
14
necropsy
lobes in separate vials,


3B
4
A277 mxRNA (30 mg/kg)
14
days 14
in RNAlater for 15 min,


3C
4
A277 mxRNA (10 mg/kg)
42
and 42.
flash freeze then


Spares
4


Send
stored at −80° C.


Total
36


Plasma and
2 mm biopsy of left,






serum to
middle and right liver






Sponsor.
all in one vial, flash







freeze then stored 







at −80° C.







Rest of liver, flash







freeze then stored 







at −80° C.









Prior to necropsy, the animals will be deeply anesthetized and a terminal blood draw will be performed through the vena cava. The target blood volume to be collected per animal is as much blood as possible with a minimum of 1.2 mL which will be split equally between a serum and plasma separation tube. After separation (see section 14.10) the serum will be split equally in two separate vials and plasma also will be separated in two separate vials (see example below).

    
    
        1.2 mL of blood=0.6 mL for serum and 0.6 mL for plasma separation tubes
        Serum (0.3 mL after separation)=0.15 mL×2 vials
        Plasma (0.3 mL after separation)=0.15 mL×2 vials.
    
    


Above serum and plasma samples will be labelled, flash frozen and stored at −80° C.
Additional blood collected over the minimum 1.2 mL volume will be placed in a serum separation tube, processed, serum transferred to a labelled vile, refrigerated at 4° C. for rodent lipid analysis.
Note: serum and plasma will be used to measure protein, caution should be taken to avoid hemolysis or clot formation.
At necropsy, three 2 mm biopsy punches will be taken from the left, middle and right liver lobes, placed in separate vials, soaked in RNAlater for 15 minutes, flash frozen and stored at −80° C. Another three 2 mm liver biopsies from the left, middle and right liver lobes will be placed into one vial, flash frozen and stored at −80° C. The rest of the liver will be flash frozen and stored in 10 mL conical tubes at −80° C.
Alteration of Study Design
Alterations of this protocol may be made as the study progresses. Changes (to the protocol) that have the potential to negatively impact the study or the safety of the study subjects would require IACUC approval.
Animal Inclusion and Exclusion Criteria
Any animals that are deemed unhealthy during veterinary pre-screen will be excluded from the study and replaced with a spare animal if available. For survival animals found dead or moribund after treatment may be replaced via study protocol amendment by a spare animal if available.
Animal Disposition
At the end of the study, the animals will be euthanized.
Route of Administration
Subcutaneous injection in the scruff. An injection volume of 200 uL.
Results
FIG. 3 highlights the dose-response effect on the percent reduction of APOC3 mRNA in the liver tissues and APOC3 protein levels in the plasma of the animals treated with the different mxRNA constructs at Day 14 as compared to the control animals.
In addition, the following notes apply to FIG. 3:

    
    
        A28(14-4)mF-10=A28(14-4)mF 10 mg/kg dose group
        A28(14-4)mF-30=A28(14-4)mF 30 mg/kg dose group
        A277(12-5)-10=A277(12-5) 10 mg/kg dose group
        A277(12-5)-30=A277(12-5) 30 mg/kg dose group FIG. 4 highlights the dose-response effect on the mean percent reduction of Triglycerides and Total Cholesterol in the serum of the animals treated with the different APOC-3 targeting mxRNA constructs at Day 14 as compared to the control animals.
    
    


FIGS. 5a and 5b highlight the duration effect on the mean percent reduction of APOC3 mRNA in liver tissues and APOC3 protein levels in the plasma of the animals treated with the different APOC3-targeting mxRNA (10 mg/kg) constructs at Day 14 (Week 2) and at Week 6 as compared to the control animals. Moreover, it is noted with respect to these Figures that an outlier from the A277(12-5) group is excluded.
FIGS. 6a and 6b highlight the duration effect on the mean percent reduction of triglycerides (TGs) and total cholesterol (TC) in the serum of the animals treated with the different APOC3-targeting mxRNA (10 mg/kg) constructs at Day 14 (Week 2) and at Week 6 as compared to the control animals. With respect to these Figures it is noted, that an outlier from the A277(12-5) group is excluded.
Summary of Results
A28(14-4)mF APOC3-targeting mxRNA construct:

    
    
        88% suppression of APOC3 mRNA as compared to control group at week 2 that was maintained at 78% on Week 6.
        90% reduction in plasma APOC3 levels as compared to control group at week 6 that was sustained at 85% on Week 6.
        32% reduction in serum triglycerides levels as compared to control group at week 2 that increased to 41% reduction on Week 6.
        43% reduction in serum total cholesterol levels as compared to control group at week 2 that was maintained at 33% on Week 6.
    
    


A277(12-5) APOC3-targeting mxRNA construct:

    
    
        56% suppression of APOC3 mRNA as compared to control group at week 2 that was maintained at 42% on Week 6.
        83% reduction in plasma APOC3 levels as compared to control group at week 6 that was sustained at 84% on Week 6.
        8% reduction in serum triglycerides levels as compared to control group at week 2 that increased to 52% reduction on Week 6.
        36% reduction in serum total cholesterol levels as compared to control group at week 2 that was lost on Week 6.
    
    


CONCLUSIONS
Construct A28(14-4)mF produced outstanding activity, with 98% of the targeted protein downregulation at 2-week timepoint at 30 mg/kg dosing. Furthermore, construct A28(14-4)mF sustained excellent (protein knockdown) activity at 10 mg/kg dosing both on week 2 and week 6.
Example 5
Following the protocol described in detail in Example 4, the effects of compound A28(14-4)mF (also designated STP125G) have been observed over a longer period of time. See FIG. 7 for an overview of this extended study. The corresponding results are displayed in FIGS. 8a and 8b (APOC3 mRNA and protein knockdown, respectively), and FIGS. 9a and 9b (triglyceride and total cholesterol levels).
Several Aspects are Notable:


    
    
        A single dose of 10 mg/kg is sufficient for knockdown of mRNA and protein for a period of six weeks with a rebound becoming slowly apparent toward the end of the study.
        Not only triglycerides (fat levels in blood primarily considered to be associated with APOC3) but also total cholesterol are downregulated.
        In the assessment of the latter findings, the properties of the mice used for the study must be considered. FIG. 10 shows that an estimated fraction of 20 to 25 percent of the cells of the humanized liver remain murine (mouse) cells. A28A(14-4)mF does not target murine APOC3. As a consequence, the non-silenced murine APOC3 contributes to the observed triglyceride and total cholesterol levels. Thus, the downregulation of these two blood fats in a purely human system is expected to exceed the results observed in this study.