TH80958A - Inhibition of eye symptoms with RNAi-mediated - Google Patents

Abstract

DC60 (29/03/49) RNA activation is used to inhibit the expression of target mRNAs for intraocular pressure to reduce elevated intraocular pressure in patients with open-angle glaucoma or hyperintraocular pressure. Targets for hyperintraocular pressure include carbonic anhydrase II, IV and renal beta-1- and beta-2- receptors; acetylcholinesterase; Na+/K+-ATPase and the Na-K-2CI cotransmitter. Such hyperintraocular pressure symptoms can be treated using the activation RNAs of this invention. RNA activation is used to inhibit the expression of target mRNAs for intraocular pressure to reduce elevated intraocular pressure in patients with open-angle glaucoma or hyperintraocular pressure. Targets for hyperintraocular pressure include carbonic anhydrase II, IV and renal beta-1- and beta-2- receptors; acetylcholinesterase; The Na+/K+- ATPase and its co-transmitter Na-K-2Cl can treat this hyperocular pressure condition using the activation RNAs of this invention.

Claims (33)

1. Components for reducing the expression of target mRNA for hypertension in the eye. This component consists of stimulating RNA of 19 to 49 nucleotides and a drug-tolerant vector. The stimulating RNA contains a sensory nucleotide strand, an antisensory nucleotide strand, and at least a nearly complete complement region containing at least 19 nucleotides, where the antisensory strand induces recombination under physiological conditions depending on the mRNA region corresponding to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:101, or SEQ ID NO:134, and the nearly complete complement region containing at least 19 nucleotides with the mRNA recombinant region corresponding to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:101, or SEQ ID NO:134, respectively. 2. The component according to claim 1, where the target mRNA for high intraocular pressure encodes carbonic hydrase II and the anti-sensing strand generates a recombinant under physiological conditions, depends on the portion of mRNA matching SEQ ID NO:1 and has at least a nearly complete complementary region containing at least 19 nucleotides with the recombinant portion of mRNA matching SEQ ID NO:1. 3. The component according to claim 1, where the intraocular pressure target mRNA encodes carbonic hydrase IV and the anti-sensing strand generates a recombinant under physiological conditions, depends on the mRNA segment matching SEQ ID NO:2 and has at least a nearly complete complementary region containing at least 19 nucleotides with the recombinant mRNA segment matching SEQ ID NO:2. 4. The component according to claim 1, where the target mRNA for high intraocular pressure encodes carbonic anhydrase XII and the antispasmodic strand produces a hybrid under physiological conditions depending on the mRNA region corresponding to SEQ ID NO:101, or SEQ ID NO:134, and has at least a nearly complete complement region of at least 19 nucleotides with the hybrid mRNA region corresponding to SEQ ID NO:101, or SEQ ID NO:134, respectively. 5. Component according to claim 1, where the antisensitizing strand is designed to target mRNA matching SEQ ID NO:1 containing nucleotides 232,527,721, 728,809,810,855,856,921,1139,506,547,548,740,911,1009,1140,1149,1150, 1151,1188,1194,1195,1223,1239,1456,1457,1458,100,158,166,247,286,318, 322,328,371,412,482,504,505,541,734,772,777,814,972,998. Or 1232 6. Component according to Claim 1 where the antisensitizing strand is designed to target mRNA matching SEQ ID NO:2 containing nucleotides 213, 252, 258, 266, 399, 457, 463, 490, 595, 1064, 109, 112, 125, 126, 150, 261, 265, 280, 398, 453, 459, 462, 457, 492, 534, 785, 801, 825, 827, 876, 1003, or 1012. 7. Component according to Claim 1 where the antisensitizing strand is designed to target mRNA matching SEQ ID NO:101 containing nucleotides 191,239,274, 275,341,389,412,413,423,687,689,695,710,791,792,794,983,993,994,995,691, 1039,1568,2326,2332,2425,2433,2844,2845,2880,2884,2891,2954,2955,2956, 2957,2964,2965,3006,3007,3012, or 3026. 8. Component according to Claim 1 where the antisensitizing strand is designed to target mRNA matching SEQ ID NO:134 containing nucleotides 687, 1535, 2293, 2299, 2392, 2400, 2811, 2812, 2847, 2851, 2858, 2921, 2922, 2923, 2924, 2931, 2932, 2973, 2974, 2979, or 2993. 9. The component according to claim 1 also contains a second activation RNA of 19 to 49 nucleotides in length, comprising a sensory nucleotide strand, an antisensory nucleotide strand, and at least a nearly complete complement region containing at least 19 nucleotides, where the antisensory strand fuses under physiological conditions with the mRNA region corresponding to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 101, or SEQ ID NO: 134, and has at least a nearly complete complement region containing at least 19 nucleotides with the mRNA fuse region corresponding to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 101, or SEQ ID NO: 134, respectively. 10. The element according to claim 1 where the sensory nucleotide strand and the antisensory nucleotide strand are connected by a loop nucleotide series. 11. Method for reducing the expression of variant one of the target mRNA for hyperintraocular pressure without reducing the expression of variant two of the target mRNA for hyperintraocular pressure in two test samples. This method consists of the following components: stimulating RNA of 19 to 49 nucleotides in length in an effective amount and a suitable vector for the test sample. The stimulating RNA contains a continuous region of at least 13 nucleotides with at least 90% serial complement or at least 90% serial identity with 13 nucleotides from the left of the 3' end of variant one, where the expression of variant one mRNA is reduced without a reduction in the expression of variant two mRNA, and where the target mRNA of variant one is SEQ ID NO:101 and the target mRNA of variant two is SEQ ID NO:134. 12. Method for reducing the expression of variant one of the target mRNA for hyperintraocular pressure without reducing the expression of variant two of the target mRNA for hyperintraocular pressure in test samples. This method involves using the following components: stimulating RNA of 19 to 49 nucleotides in an effective amount and a drug-tolerant vector for the test sample. The stimulating RNA contains a continuous region of at least 13 nucleotides with at least 90% serial complement or at least 90% serial identity with 13 nucleotides from the left of the 3' end of variant one, where the expression of variant one mRNA is reduced without a reduction in the expression of variant two mRNA, and where the target mRNA of variant one is SEQ ID NO:134 and the target mRNA of variant two is SEQ ID NO:101. 13. Component for reducing the expression of target mRNA in hypertensive intraocular hyperplasia. This component consists of stimulating RNA of 19 to 49 nucleotides in length and a drug-tolerant vector, where the single-stranded stimulating RNA induces recombination under certain conditions. Physiologically, it depends on the mRNA segment corresponding to SEQ ID NO:1, which consists of 232, 527,721,728,809,810,855,856,921,1139,506,547,548,740,911,1009,1140,1149, 1150,1151,1188,1194,1195,1223,1239,1456,1457,1458,100,158,166,247,286, 318,322,328,371,412,482,504,505,514,734,772,777,814,972,998, or 1232 and RNA. The stimulus RNA has at least a nearly complete complement region with the recombinant mRNA region corresponding to SEQ ID NO:1, where the single-stranded stimulus RNA induces recombination under physiological conditions depending on the mRNA region corresponding to SEQ ID NO:2, which includes 213,252, 258,266,399,457,463,490,595,1064,109,112,125,126,150,261,265,280,398, 453,459,462,467,492,534,785,801,825,827,876,1003, or 1012, and the stimulus RNA has at least a nearly complete complement region with the recombinant mRNA region corresponding to SEQ ID NO:2, or where the single-stranded stimulus RNA induces recombination under physiological conditions. The physiological state depends on the mRNA region corresponding to SEQ ID NO:101, which includes 191,239,274,275,341,389,412,413,423,687,689,695,710,791,792,794,983,993, 994,995,691,1039,1568,2326,2332,2425,2433,2844,2845,2880,2884,2891,2954, 2955,2956,2957,2964,2965,3006,3007,3012, or 3026, and the stimulating RNA region. A region of at least near-complete continuum complement with the mRNA recombinant region corresponding to SEQ ID NO:101, or where a single-stranded activation RNA strand induces a recombination under physiologically dependent conditions with the mRNA region corresponding to SEQ ID NO:134 containing 687,1535,2293,2299,2392,2400,2811,2812,2847,2851,2858,2921,2922,2923, 2924,2931,2932,2973,2974,2979, or 2993, and the activation RNA has at least a region of near-complete continuum complement with the mRNA recombinant region corresponding to SEQ ID NO:134. 14. Components for reducing the expression of target mRNA for hypertension in the eye, consisting of stimulating RNA of 19 to 49 nucleotides in length and a drug-tolerant vector. The stimulating RNA contains a continuous region of at least 13 nucleotides with at least 90% serial complementarity or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:8, SEQ ID NO:14 - SEQ ID NO:32, SEQ ID NO:83 - SEQ ID NO:100, SEQ ID NO:102 - SEQ ID NO:122, and SEQ ID NO:135 - SEQ ID NO:219. 15. Components commensurate with claim 14 where the intraocular hypertension target mRNA encodes carbonic anhydrase II and stimulating RNA contains a continuous nucleotide region of at least 13 nucleotides with at least 90% serial complementarity or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:8, SEQ ID NO:14 - SEQ ID NO:22, SEQ ID NO:83 - SEQ ID NO:100, SEQ ID NO:102 - SEQ ID NO:122, and SEQ ID NO:135 - SEQ ID NO:155. 16. Components stipulated in claim 14 where the intraocular hypertensive target mRNA encodes carbonic anhydrase IV and stimulating RNA containing a continuous nucleotide region of at least 13 nucleotides with at least 90% serial complementarity or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any SEQ ID NO:23 - SEQ ID NO:32, SEQ ID NO:156 - SEQ ID NO:177. 17. Components stipulated in claim 14 where the target mRNA for intraocular hypertension encoding carbonic anhydrase XII and the stimulating RNA containing a continuous nucleotide region of at least 13 nucleotides with at least 90% serial complementarity or at least 90% serial identity with 13 nucleotides from the left of the 3' end of either SEQ ID NO:102 - SEQ ID NO:122, and SEQ ID NO:178 - SEQ ID NO:219. 18. An element pursuant to claim 14 where the nucleotide continuum is a nucleotide continuum of 14 nucleotides with at least 85% series complement or at least 85% series identity with 14 nucleotides from the left of the 3' end of the series identifier. 19. An element pursuant to claim 14 where the nucleotide continuum is a nucleotide continuum of 15, 16, 17, or 18 nucleotides with at least 85% series complement or at least 85% series identity with 15, 16, 17, or 18 nucleotides from the left of the 3' end of the series identifier. 20. The element according to claim 14 where the RNA activates the sHRNA. 21. The claimed components are stimulating RNA of 19 to 49 nucleotides in length and belonging to one of the following nucleotide series: SEQ ID NO:8, SEQ ID NO:14 - SEQ ID NO:32, SEQ ID NO:83 - SEQ ID NO:100, SEQ ID NO:102 - SEQ ID NO:122, and SEQ ID NO:135 - SEQ ID NO:219, or an additive to such series, and a pharmacologically acceptable vector. 22. The element according to claim 21 where the stimulating RNA is sHRNA. 23. The use of this component involves an effective and pharmacologically acceptable stimulating RNA of 19 to 49 nucleotides in the form of a drug for the treatment of intraocular hypertension in the test sample. The stimulating RNA contains a continuous region of at least 13 nucleotides with at least 90% serial complementarity or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:8, SEQ ID NO:14 - SEQ ID NO:22, SEQ ID NO:83 - SEQ ID NO:100, SEQ ID NO:102 - SEQ ID NO:122, and SEQ ID NO:135 - SEQ ID NO:155. 24. Use of elements according to claim 23 where the stimulating RNA contains a continuous nucleotide region of at least 13 nucleotides with at least 90% serial complement or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:8, SEQ ID NO:14 - SEQ ID NO:22, SEQ ID NO:83 - SEQ ID NO:100, and SEQ ID NO:135 - SEQ ID NO:155. 25. Use of elements according to claim 23 where the stimulating RNA contains a continuous nucleotide region of at least 13 nucleotides with at least 90% serial complement or at least 90% serial identity with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:23, SEQ ID NO:35 and SEQ ID NO:156 - SEQ ID NO:177. 26. Use of elements according to claim 23 where the stimulating RNA contains at least 90% continuous nucleotide regions, at least 90% serial complement nucleotides to, or at least 90% serial characteristic with 13 nucleotides from the left of the 3' end of any one of SEQ ID NO:102 - SEQ ID NO:122 and SEQ ID NO:178 - SEQ ID NO:219. 27. Using the element according to claim 23, where the stimulating RNA is sHRNA. 28. Use of the components under claim 23 where such components are prepared for topical use in the eye, through the eye, around the eye, eyelid membrane, subjunctivitis, in the lens, under the retina, under the conjunctiva, behind the eyeball, or in the retina. 29. Utilization of components under claim 23, where such components are prepared for use through live body expression from activated RNA expression vectors. 30. This component consists of an activation RNA of 19 to 49 nucleotides in an effective and pharmacologically acceptable quantity for the preparation of a drug to treat intraocular hypertension in the test sample. The activation RNA contains a sensory nucleotide strand, an antisensory nucleotide strand, and at least a nearly complete complement region containing at least 19 nucleotides, where the antisensory strand induces recombination under physiological conditions depending on the mRNA region corresponding to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:101, or SEQ ID NO:134, and at least a nearly complete complement region containing at least 19 nucleotides with the mRNA recombinant region corresponding to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:101, or SEQ ID NO:134, respectively. 31. Utilizing the elements according to claim 30, where the sensory nucleotide chain and the antisensory nucleotide chain are connected by a loop nucleotide series. 32. Use of the components under claim 30, where such components are prepared for topical use in the eye, through the eye, around the eye, eyelid membrane, subjunctivitis, in the lens, under the retina, under the conjunctiva, behind the eyeball, or in the retina. 33. Utilization of components under claim 30, where such components are prepared for use through in vivo expression from activated RNA expression vectors.