Rnas for wound healing

Abstract

The present invention relates to an RNA encoding a therapeutic protein, in particular a collagenase, growth factor, cytokine, receptor, chaperone or signal transduction inhibitor. In particular, the present invention relates to RNA suitable for treatment of wounds, specifically for promoting wound healing. The present invention concerns such RNA as well as pharmaceutical compositions and kits and combinations comprising the RNA. Furthermore, the present invention relates to the RNA, pharmaceutical compositions, kits as disclosed herein for use in the treatment of wounds, specifically for promoting wound healing.

1 . A method of treating a wound in a subject in need thereof comprising administering to the subject an effective amount of RNA comprising at least one coding sequence, wherein the coding sequence encodes at least one therapeutic protein selected from a collagenase selected from MMP1; ColG; ColH; ColG/ColH mixture; MMP8; MMP9; MMP13; a growth factor selected from HGF; VEGFA; FGF21; AMELX; AMELY; ssAMELX; ssAMELX-001-1; ssAMELX-001-2; ssAMELX-002; ssAMELX-003; ssAMELX-004; ssAMELX-201; BMP1; BMP2; BMP4; BMP6; BMP7; EGF; EREG; FGF1; FGF2; FGF7; HBEGF; IGF1; IGF2; INHBA; INHBB; PDGFA; PDGFB; PDGFC; PDGFD; TGFA; TGFB1; TGFB2; TGFB3; PGF; VEGFB; VEGFC or VEGFD; a cytokine selected from IL6 or CCL7; a receptor selected from ITGAM, CCR1 or TNFRSF1B; a chaperone selected from HSPA1A; HSPA1B; HSPA1L; HSPA2; HSPA4; HSPA4L; HSPA5; HSPA6; HSPA7; HSPA8; HSPA9; HSPA12A; HSPA12B; HSPA13; HSPA14; HSPH1; HSP90AA1; HSP90AA3P; HSP90AB1; HSP90B1; HYOU1, extracellular matrix protease or TRAP1; or SOCS3. 
     
     
         2 . The method of  claim 1 , wherein said RNA comprises at least one coding sequence, wherein the coding sequence encodes a therapeutic protein selected from a collagenase, preferably selected from MMP1; ColG or ColH. 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein the therapeutic protein comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 6, 7; 8; 9; 10; 11; 12; 13; 14; 15; 16; 17; 18; 19; 20; 21; 22; 23; 24; 25; 26; 27; 28; 29; 30; 31; 32; 33; 34; 35; 36; 37; 38; 39; 40; 41; 42; 43; 44; 45; 46; 47; 48; 49; 50; 51; 52; 53; 54; 55; 56; 57; 58; 59; 60; 61; 62; 63; 64; 65; 66; 67; 68; 69; 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80; 81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 5192; 5193; 5194 or 5195. 
     
     
         5 . The method of  claim 1 , wherein said at least one coding region comprises or consists of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 104; 105; 106; 107; 108; 109; 110; 111; 112; 113; 114; 115; 116; 117; 118; 119; 120; 121; 122; 123; 124; 125; 126; 127; 128; 129; 130; 131; 132; 133; 134; 135; 136; 137; 138; 139; 140; 141; 142; 143; 144; 145; 146; 147; 148; 149; 150; 151; 152; 153; 154; 155; 156; 157; 158; 159; 160; 161; 162; 163; 164; 165; 166; 167; 168; 169; 170; 171; 172; 173; 174; 175; 176; 177; 178; 179; 180; 181; 182; 183; 184; 185; 186; 187; 188; 189; 190; 191; 192; 193; 194; 195; 196; 197; 198; 199; 200; 201; 202; 203; 204; 205; 206; 207; 208; 209; 210; 216; 217; 218; 219; 220; 221; 222; 223; 224; 225; 226; 227; 228; 229; 230; 231; 232; 233; 234; 235; 236; 237; 238; 239; 240; 241; 242; 243; 244; 245; 246; 247; 248; 249; 250; 251; 252; 253; 254; 255; 256; 257; 258; 259; 260; 261; 262; 263; 264; 265; 266; 267; 268; 269; 270; 271; 272; 273; 274; 275; 276; 277; 278; 279; 280; 281; 282; 283; 284; 285; 286; 287; 288; 289; 290; 291; 292; 293; 294; 295; 296; 297; 298; 299; 300; 301; 302; 303; 304; 305; 306; 307; 313; 314; 315; 316; 317; 318; 319; 320; 321; 322; 323; 324; 325; 326; 327; 328; 329; 330; 331; 332; 333; 334; 335; 336; 337; 338; 339; 340; 341; 342; 343; 344; 345; 346; 347; 348; 349; 350; 351; 352; 353; 354; 355; 356; 357; 358; 359; 360; 361; 362; 363; 364; 365; 366; 367; 368; 369; 370; 371; 372; 373; 374; 375; 376; 377; 378; 379; 380; 381; 382; 383; 384; 385; 386; 387; 388; 389; 390; 391; 392; 393; 394; 395; 396; 397; 398; 399; 400; 401; 402; 403; 404; 405, 406, 407, 408, 409, 410; 411; 412; 413; 414; 415; 416; 417; 418; 419; 420; 421; 422; 423; 424; 425; 426; 427; 428; 429; 430; 431; 432; 433; 434; 435; 436; 437; 438; 439; 440; 441; 442; 443; 444; 445; 446; 447; 448; 449; 450; 451; 452; 453; 454; 455; 456; 457; 458; 459; 460; 461; 462; 463; 464; 465; 466; 467; 468; 469; 470; 471; 472; 473; 474; 475; 476; 477; 478; 479; 480; 481; 482; 483; 484; 485; 486; 487; 488; 489; 490; 491; 492; 493; 494; 495; 496; 497; 498; 499; 500; 501; 507; 508; 509; 510; 511; 512; 513; 514; 515; 516; 517; 518; 519; 522; 521; 522; 523; 524; 525; 526; 527; 528; 529; 530; 531; 532; 533; 534; 535; 536; 537; 538; 539; 540; 541; 542; 543; 544; 545; 546; 547; 548; 549; 550; 551; 552; 553; 554; 555; 556; 557; 558; 559; 560; 561; 562; 563; 564; 565; 566; 567; 568; 569; 570; 571; 572; 573; 574; 575; 576; 577; 578; 579; 580; 581; 582; 583; 584; 585; 586; 587; 588; 589; 590; 591; 592; 593; 594; 595; 596; 597; 598; 604; 605; 606; 607; 608; 609; 610; 611; 612; 613; 614; 615; 616; 617M 618; 619; 620; 621; 622; 623; 624; 625; 626; 627; 628; 629; 630; 631; 632; 633; 634; 635; 636; 637; 638; 639; 640; 641; 642; 643; 644; 645; 646; 647; 648; 649; 650; 651; 652; 653; 654; 655; 656; 657; 658; 659; 660; 661; 662; 663; 664; 665; 666; 667; 668; 669; 670; 671; 672; 673; 674; 675; 676; 677; 678; 679; 680; 681; 682; 683; 684; 685; 686; 687; 688; 689; 690; 691; 692; 693; 694; 695; 701; 702; 703; 704; 705; 706; 707; 708; 709; 710; 711; 712; 713; 714; 715; 716; 717; 718; 719; 720; 721; 722; 723; 724; 725; 726; 727; 728; 729; 730; 731; 732; 733; 734; 735; 736; 737; 738; 739; 740; 741; 742; 743; 744; 745; 746; 747; 748; 749; 750; 751; 752; 753; 754; 755; 756; 757; 758; 759; 760; 761; 762; 763; 764; 765; 766; 767; 768; 769; 770; 771; 772; 773; 774; 775; 776; 777; 778; 779; 780; 781; 782; 783; 784; 785; 786; 787; 788; 789; 790; 791; 792; 798; 799; 800; 801; 802; 803; 804; 805; 806; 807; 808; 809; 810; 811; 812; 813; 814; 815; 816; 817; 818; 819; 820; 821; 822; 823; 824; 825; 826; 827; 828; 829; 830; 831; 832; 833; 834; 835; 836; 837; 838; 839; 840; 841; 842; 843; 844; 845; 846; 847; 848; 849; 850; 851; 852; 853; 854; 855; 856; 857; 858; 859; 860; 861; 862; 863; 864; 865; 866; 867; 868; 869; 870; 871; 872; 873; 874; 875; 876; 877; 878; 879; 880; 881; 882; 883; 884; 885; 886; 887; 888; 889; 895; 896; 897; 898; 899; 900; 901; 902; 903; 904; 905; 906; 907; 908; 909; 910; 911; 912; 913; 914; 915; 916; 917; 918; 919; 920; 921; 922; 923; 924; 925; 926; 927; 928; 929; 930; 931; 932; 933; 934; 935; 936; 937; 938; 939; 940; 941; 942; 943; 944; 945; 946; 947; 948; 949; 950; 951; 952; 953; 954; 955; 956; 957; 958; 959; 960; 961; 962; 963; 964; 965; 966; 967; 968; 969; 970; 971; 972; 973; 974; 975; 976; 977; 978; 979; 980; 981; 982; 983; 984; 985; 986; 992; 993; 994; 995; 996; 997; 998; 999; 1000; 1001; 1002; 1003; 1004; 1005; 1006; 1007; 1008; 1009; 1010; 1011; 1012; 1013; 1014; 1015; 1016; 1017; 1018; 1019; 1020; 1021; 1022; 1023; 1024; 1025; 1026; 1027; 1028; 1029; 1030; 1031; 1032; 1033; 1034; 1035; 1036; 1037; 1038; 1039; 1040; 1041; 1042; 1043; 1044; 1045; 1046; 1047; 1048; 1049; 1050; 1051; 1052; 1053; 1054; 1055; 1056; 1057; 1058; 1059; 1060; 1061; 1062; 1063; 1064; 1065; 1066; 1067; 1068; 1069; 1070; 1071; 1072; 1073; 1074; 1075; 1076; 1077; 1078; 1079; 1080; 1081; 1082; 1083; 1089; 1090; 1091; 1092; 1093; 1094; 1095; 1096; 1097; 1098; 1099; 1100; 1101; 1102; 1103; 1104; 1105; 1106; 1107; 1108; 1109; 1110; 1111; 1112; 1113; 1114; 1115; 1116; 1117; 1118; 1119; 1120; 1121; 1122; 1123; 1124; 1125; 1126; 1127; 1128; 1129; 1130; 1131; 1132; 1133; 1134; 1135; 1136; 1137; 1138; 1139; 1140; 1141; 1142; 1143; 1144; 1145; 1146; 1147; 1148; 1149; 1150; 1151; 1152; 1153; 1154; 1155; 1156; 1157; 1158; 1159; 1160; 1161; 1162; 1163; 1164; 1165; 1166; 1167; 1168; 1169; 1170; 1171; 1172; 1173; 1174; 1175; 1176; 1177; 1178; 1179; 1180; 1186; 1187; 1188; 1189; 1190; 1191; 1192; 1193; 1194; 1195; 1196; 1197; 1198; 1199; 1200; 1201; 1202; 1203; 1204; 1205; 1206; 1207; 1208; 1209; 1210; 1211; 1212; 1213; 1214; 1215; 1216; 1217; 1218; 1219; 1220; 1221; 1222; 1223; 1224; 1225; 1226; 1227; 1228; 1229; 1230; 1231; 1232; 1233; 1234; 1235; 1236; 1237; 1238; 1239; 1240; 1241; 1242; 1243; 1244; 1245; 1246; 1247; 1248; 1249; 1250; 1251; 1252; 1253; 1254; 1255; 1256; 1257; 1258; 1259; 1260; 1261; 1262; 1263; 1264; 1265; 1266; 1267; 1268; 1269; 1270; 1271; 1272; 1273; 1274; 1275; 1276; 1277; 1283; 1284; 1285; 1286; 1287; 1288; 1289; 1290; 1291; 1292; 1293; 1294; 1295; 1296; 1297; 1298; 1299; 1300; 1301; 1302; 1303; 1304; 1305; 1306; 1307; 1308; 1309; 1310; 1311; 1312; 1313; 1314; 1315; 1316; 1317; 1318; 1319; 1320; 1321; 1322; 1323; 1324; 1325; 1326; 1327; 1328; 1329; 1330; 1331; 1332; 1333; 1334; 1335; 1336; 1337; 1338; 1339; 1340; 1341; 1342; 1343; 1344; 1345; 1346; 1347; 1348; 1349; 1350; 1351; 1352; 1353; 1354; 1355; 1356; 1357; 1358; 1359; 1360; 1361; 1362; 1363; 1364; 1365; 1366; 1367; 1368; 1369; 1370; 1371; 1372; 1373; 1374; 5196; 5197; 5198; 5199; 5200; 5201; 5202; 5203; 5204; 5205; 5206; 5207; 5208; 5209; 5210; 5211; 5212; 5213; 5214; 5215; 5216; 5217; 5218; 5219; 5220; 5221; 5222; 5223; 5224; 5225; 5226; 5227; 5228; 5229; 5230; 5231; 5232; 5233; 5234; 5235; 5236; 5237; 5238; 5239; 5240; 5241; 5242; 5243; 5244; 5245; 5246; 5247; 5248; 5249; 5250 or 5251. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the RNA is mono-, bi-, or multicistronic. 
     
     
         8 . The method of  claim 1 , wherein the RNA is an mRNA, a viral RNA or a replicon RNA. 
     
     
         9 . The method of  claim 1 , wherein the RNA is a modified RNA, preferably a stabilized RNA. 
     
     
         10 . The method of  claim 1 , wherein
 a) the G/C content of the at least one coding sequence of the RNA is increased compared to the G/C content of the corresponding coding sequence of the corresponding wild type RNA, and/or wherein   b) the C content of the at least one coding sequence of the RNA is increased compared to the C content of the corresponding coding sequence of the corresponding wild type RNA, and/or wherein   c) the codons in the at least one coding sequence of the RNA are adapted to human codon usage, wherein the codon adaptation index (CAI) is preferably increased or maximised in the at least one coding sequence of the RNA,   d) wherein the amino acid sequence encoded by the RNA is preferably not being modified compared to the amino acid sequence encoded by the corresponding wild type RNA.   
     
     
         11 . The method of  claim 1 , which comprises a 5′-cap structure and/or at least one 3′-untranslated region element (3′-UTR element). 
     
     
         12 . The method of  claim 1 , which comprises at least one histone stem-loop. 
     
     
         13 - 15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein the at least one RNA comprises a poly(A) sequence; comprising 10 to 200, 10 to 100, 40 to 80 or 50 to 70 adenosine nucleotides, and/or a poly(C) sequence comprising 10 to 200, 10 to 100, 20 to 70, 20 to 60 or 10 to 40 cytosine nucleotides. 
     
     
         17 . The method of  claim 1 , wherein the RNA comprises, in 5′ to 3′ direction, the following elements:
 a) a 5′-cap structure, preferably m7GpppN, 
 b) optionally a 5′-UTR element 
 b) the at least one coding sequence, 
 c) optionally a 3′-UTR element 
 d) a poly(A) tail, preferably consisting of 10 to 200, 10 to 100, 40 to 80 or 50 to 70 adenosine nucleotides, 
 e) optionally a poly(C) tail, preferably consisting of 10 to 200, 10 to 100, 20 to 70, 20 to 60 or 10 to 40 cytosine nucleotides, and 
 f) optionally a histone stem-loop, preferably comprising the RNA sequence according to SEQ ID NO: 5035. 
 
     
     
         18 . The method of  claim 1 , which comprises a 3′-UTR element and wherein the 3′-UTR element comprises a nucleic acid sequence derived from a 3′-UTR of a gene. 
     
     
         19 . The method of  claim 18 , wherein the 3′-UTR element comprises a nucleic acid sequence derived from a 3′-UTR of a gene selected from the group consisting of an albumin gene, an α-globin gene, a β-globin gene, a tyrosine hydroxylase gene, a lipoxygenase gene, and a collagen alpha gene. 
     
     
         20 - 22 . (canceled) 
     
     
         23 . The method of  claim 1 , wherein the RNA comprises a 5′-UTR element. 
     
     
         24 . The method of  claim 23 , wherein the 5′-UTR element comprises a nucleic acid sequence, which is derived from the 5′-UTR of a TOP gene. 
     
     
         25 - 30 . (canceled) 
     
     
         31 . The method of  claim 1 , wherein the at least one coding sequence encodes a collagenase. 
     
     
         32 . The method of  claim 1 , comprising administering at least one coding sequence encoding a collagenase in combination with an RNA comprising at least one coding sequence encoding a growth factor. 
     
     
         33 . A composition comprising the RNA and a pharmaceutically acceptable carrier, said RNA comprising at least one coding sequence, wherein the coding sequence encodes at least one therapeutic protein selected from a collagenase selected from MMP1; ColG; ColH; ColG/ColH mixture; MMP8; MMP9; MMP13; a growth factor selected from HGF; VEGFA; FGF21; AMELX; AMELY; ssAMELX; ssAMELX-001-1; ssAMELX-001-2; ssAMELX-002; ssAMELX-003; ssAMELX-004; ssAMELX-201; BMP1; BMP2; BMP4; BMP6; BMP7; EGF; EREG; FGF1; FGF2; FGF7; HBEGF; IGF1; IGF2; INHBA; INHBB; PDGFA; PDGFB; PDGFC; PDGFD; TGFA; TGFB1; TGFB2; TGFB3; PGF; VEGFB; VEGFC or VEGFD; a cytokine selected from IL6 or CCL7; a receptor selected from ITGAM, CCR1 or TNFRSF1B; a chaperone selected from HSPA1A; HSPA1B; HSPA1L; HSPA2; HSPA4; HSPA4L; HSPA5; HSPA6; HSPA7; HSPA8; HSPA9; HSPA12A; HSPA12B; HSPA13; HSPA14; HSPH1; HSP90AA1; HSP90AA3P; HSP90AB1; HSP90B1; HYOU1, extracellular matrix protease or TRAP1; or SOCS3. 
     
     
         34 - 48 . (canceled) 
     
     
         49 . A wound dressing comprising a RNA, said RNA comprising at least one coding sequence, wherein the coding sequence encodes at least one therapeutic protein selected from a collagenase selected from MMP1; ColG; ColH; ColG/ColH mixture; MMP8; MMP9; MMP13; a growth factor selected from HGF; VEGFA; FGF21; AMELX; AMELY; ssAMELX; ssAMELX-001-1; ssAMELX-001-2; ssAMELX-002; ssAMELX-003; ssAMELX-004; ssAMELX-201; BMP1; BMP2; BMP4; BMP6; BMP7; EGF; EREG; FGF1; FGF2; FGF7; HBEGF; IGF1; IGF2; INHBA; INHBB; PDGFA; PDGFB; PDGFC; PDGFD; TGFA; TGFB1; TGFB2; TGFB3; PGF; VEGFB; VEGFC or VEGFD; a cytokine selected from IL6 or CCL7; a receptor selected from ITGAM, CCR1 or TNFRSF1B; a chaperone selected from HSPA1A; HSPA1B; HSPA1L; HSPA2; HSPA4; HSPA4L; HSPA5; HSPA6; HSPA7; HSPA8; HSPA9; HSPA12A; HSPA12B; HSPA13: HSPA14; HSPH1; HSP90AA1; HSP90AA3P; HSP90AB1; HSP90B1; HYOU1, extracellular matrix protease or TRAP1; or SOCS3.