RNA interference compositions and methods for malignant tumors

Patent metadata

Field	Value
Publication number	US-RE48887-E
Application number	US-201916576102-A
Country	US
Kind code	E1
Filing date	Sep 19, 2019
Priority date	Dec 26, 2014
Publication date	Jan 11, 2022
Grant date	Jan 11, 2022

Abstract

This invention provides compositions for use in distributing active agents for treating a malignant tumor in a subject. The compositions contain RNAi molecules targeted to a human GST-π, along with RNAi molecules targeted to a human p21, and a pharmaceutically acceptable carrier. The carrier can include nanoparticles composed of an ionizable lipid, a structural lipid, one or more stabilizer lipids, and a lipid for reducing immunogenicity of the nanoparticles. This invention further provides methods for preventing or treating a malignant tumor by administering a therapeutically effective amount of an RNAi composition.

First claim

What is claimed is:

1. A composition comprising RNAi molecules targeted to a human GST-π, RNAi molecules targeted to a human p21, and a pharmaceutically acceptable carrier, wherein each of the RNAi molecules targeted to GST-π has an antisense strand SEQ ID NOs:131 and a sense strand SEQ ID NOs: 157.

2. The composition of claim 1 , wherein each of the RNAi molecules targeted to p21 has an antisense strand SEQ ID NOs:341 and a sense strand SEQ ID NOs:355.

3. The composition of claim 1 , wherein each of the RNAi molecules targeted to GST-π has an antisense strand SEQ ID NOs:156 and a sense strand SEQ ID NOs:182.

4. The composition of claim 2 , wherein each of the RNAi molecules targeted to p21 has an antisense strand SEQ ID NOs:343 and a sense strand SEQ ID NOs:357.

5. The composition of claim 1 , wherein one or more of the nucleotides in the antisense strand or sense strand of the RNAi molecules is modified or chemically-modified.

6. The composition of claim 5 , wherein the modified or chemically-modified nucleotides are 2′-deoxy nucleotides, 2′-O-alkyl substituted nucleotides, 2′-deoxy-2′-fluoro substituted nucleotides, phosphorothioate nucleotides, locked nucleotides, or any combination thereof.

7. The composition of claim 1 , wherein each of the RNAi molecules contains a 2′-deoxynucleotide in one or more of positions 2 to 8 from the 5′ end of the antisense strand.

8. The composition of claim 1 , wherein the antisense strand of each of the RNAi molecules has deoxynucleotides in a plurality of positions, the plurality of positions being one of the following:
each of positions 4, 6 and 8, from the 5′ end of the antisense strand;
each of positions 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 1, 3, 5 and 7, from the 5′ end of the antisense strand;
each of positions 3-8, from the 5′ end of the antisense strand; or
each of positions 5-8, from the 5′ end of the antisense strand.

9. The composition of claim 1 , wherein the composition inhibits expression of GST-π mRNA with an IC50 of less than 50 pM, and inhibits expression of p21 mRNA with an IC50 of less than 50 pM.

10. The composition of claim 1 , wherein a single administration of the composition inhibits expression of GST-π mRNA levels in vivo by at least 25%.

11. The composition of claim 1 , wherein the carrier comprises liposome nanoparticles that encapsulate the RNAi molecules.

12. The composition of claim 1 , wherein the carrier comprises liposome nanoparticles that encapsulate the RNAi molecules and retain at least 80% of the encapsulated RNAi molecules after 1 hour exposure to human serum.

13. The composition of claim 1 , wherein the carrier comprises liposome nanoparticles having a size of 10 to 1000 nm, or 10 to 150 nm.

14. The composition of claim 1 , wherein the composition is active for treating malignant tumor.

15. The composition of claim 14 , wherein the malignant tumor is located in the lung, colon, kidney, pancreas, liver, bone, skin, or intestine.

16. The composition of claim 1 , wherein the carrier comprises liposome nanoparticles comprising an ionizable lipid, a structural lipid, one or more stabilizer lipids, and a lipid for reducing immunogenicity of the nanoparticles.

17. The composition of claim 16 , wherein the ionizable lipid is selected from the group of ((2-((3S,4R)-3,4-dihydroxypyrrolidin-1-yl)acetyl)azanediyl)bis(ethane-2,1-diyl) (9Z,9′Z,12Z,12′Z)-bis(octadeca-9,12-dienoate), ((2-(3-(hydroxymethyl)azetidin-1-yl)acetyl)azanediyl)bis (ethane-2,1-diyl) ditetradecanoate, ((2-(4-(2-hydroxyethyl)piperazin-1-yl)acetyl)azanediyl)bis(ethane-2,1-diyl) (9Z,9′Z,12Z,12′Z)-bis(octadeca-9,12-dienoate), ((2-(4-(2-hydroxyethyl) piperazin-1-yl)acetyl)azanediyl)bis(ethane-2,1-diyl) (9Z,9′Z,12Z,12′Z)-bis(octadeca-9,12-dienoate), 2-((1-(((9Z,12Z)-heptadeca-9,12-dien-1-yl)oxy)-5-(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)-1,5-dioxopentan-3-yl) amino)-N,N,N-trimethyl-2-oxoethan-1-aminium, 2-((9Z,12Z)-N-(3-(dimethylamino)propyl)octadeca-9,12-dienamido)ethyl (9Z,12Z)-octadeca-9,12-dienoate, N,N,N-trimethyl-3-((9Z,12Z)-N-(2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)ethyl)octadeca-9,12-dienamido)propan-1-aminium, and N,N,N-trimethyl-2-(((S)-3-(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)-2-((9Z,12Z)-octadeca-9,12-dienamido)-3-oxopropyl)amino)-2-oxoethan-1-aminium.

18. A method for preventing, treating or ameliorating one or more symptoms of a malignant tumor in a subject in need, the method comprising administering to the subject an effective amount of a composition of claim 1 .

19. The method of claim 18 , wherein the malignant tumor is associated with KRAS mutation, the method further comprising identifying a tumor cell in the subject, the tumor cell comprising at least one of: (i) a mutation of the KRAS gene, and (ii) an aberrant expression level of KRAS protein.

20. The method of claim 18 , wherein the malignant tumor overexpresses GST-π.

21. The method of claim 18 , wherein the RNAi molecules decrease expression of GST-π and p21 in the subject.

22. The method of claim 18 , wherein the administration decreases expression of GST-π and p21 in the subject by at least 5% for at least 5 days.

23. The method of claim 18 , wherein the administration decreases the volume of the malignant tumor in the subject by at least 5%, or at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%.

24. The method of claim 18 , wherein the method reduces one or more symptoms of the malignant tumor, or delays or terminates the progression of the malignant tumor.

25. The method of claim 18 , wherein the administration reduces growth of malignant tumor cells in the subject.

26. The method of claim 18 , wherein the administration reduces growth for at least 2%, or at least 5%, or at least 10%, or at least 15%, or at least 20% of the malignant tumor cells in the subject.

27. The method of claim 18 , wherein the malignant tumor is colon cancer, pancreatic cancer, kidney cancer, lung cancer, breast cancer, or fibrosarcoma.

28. The method of claim 18 , wherein the malignant tumor is lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas, or colorectal carcinoma.

29. The method of claim 18 , wherein the administration is performed from 1 to 12 times per day.

30. The method of claim 18 , wherein the administration is performed for a duration of 1, 2, 3, 4, 5, 6 or 7 days.

31. The method of claim 18 , wherein the administration is performed for a duration of 1, 2, 3, 4, 5, 6, 8, 10 or 12 weeks.

32. The method of claim 18 , wherein the administration is a dose of from 0.01 to 2 mg/kg of the RNAi molecules at least once per day for a period up to twelve weeks.

33. The method of claim 18 , wherein the administration provides a mean AUC(0-last) of from 1 to 1000 ug*min/mL and a mean C max of from 0.1 to 50 ug/mL for the GST-π RNAi molecule.

34. The method of claim 18 , wherein the administration provides a mean AUC(0-last) of from 1 to 1000 ug*min/mL and a mean C max of from 0.1 to 50 ug/mL for the p21 RNAi molecule.

35. The method of claim 18 , wherein the administration is intravenous injection, intradermal injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, oral, topical, infusion, or inhalation.

Assignees

• Nitto Denko Corp

Inventors

• Niitsu Yoshiro
• Minomi Kenjirou
• Majeti Bharat
• Wang Li
• Liu Jihua
• Adami Roger
• Ying Wenbin

Classifications

• A61K9/127

  Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant · CPC title

• C12N15/1137 Primary

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• A61K31/713 Primary

  Double-stranded nucleic acids or oligonucleotides · CPC title

• C12N2320/53

  reducing unwanted side-effects · CPC title

• C12N2320/30

  Special therapeutic applications · CPC title