Oligonucleotide analogues incorporating 5-aza-cytosine therein
Oligonucleotide analogs are provided that incorporate 5-aza-cytosine in the oligonucleotide sequence, e.g., in the form of 5-aza-2′-deoxycytidine (decitabine) or 5-aza-cytidine. In particular, oligonucleotide analogs rich in decitabine-deoxyguanosine islets (DpG and GpD) are provided to target the CpG islets in the human genome, especially in the promoter regions of genes susceptible to aberrant hypermethylation. Such analogs can be used for modulation of DNA methylation, such as effective inhibition of methylation of cytosine at the C-5 position. Methods for synthesizing these oligonucleotide analogs and for modulating nucleic acid methylation are provided. Also provided are phosphoramidite building blocks for synthesizing the oligonucleotide analogs, methods for synthesizing, formulating and administering these compounds or compositions to treat conditions, such as cancer and hematological disorders.
1. A pharmaceutical combination comprising:
1) a first therapeutic agent that is an isolated or synthetic dinucleotide analogue, or its salt or ester, comprising one 5-aza-cytosine as a base residue in the sequence of the dinucleotide analogue, wherein the dinucleotide analogue has a general formula of: 5′-DpG-3′ or 5′GpD-3′, wherein D is decitabine; p is a phospholinker wherein the number of phosphorus atoms in the phospholinker is one; and G is deoxyguanosine; and
2) a second therapeutic agent.
2. The pharmaceutical combination of claim 1 , wherein the dinucleotide analogue has the general formula of 5′-DpG-3′.
3. The pharmaceutical combination of claim 1 , wherein the dinucleotide analogue has the general formula of 5′GpD-3′.
4. The pharmaceutical combination of claim 1 , wherein p is a phosphorodiester linker.
5. The pharmaceutical combination of claim 1 , wherein the salt is a sodium, calcium, lithium, potassium, ammonium, or trialkylammonium salt.
6. The pharmaceutical combination of claim 1 , wherein the salt is a sodium salt.
7. The pharmaceutical combination of claim 1 , wherein the dinucleotide analogue is a compound of the formula:
wherein X + is a counter ion.
8. The pharmaceutical combination of claim 7 , wherein the counter ion is a sodium ion.
9. The pharmaceutical combination of claim 1 , wherein the second therapeutic agent is an alkylating agent, an agent that is a member of the retinoids superfamily, an antibiotic agent, a hormonal agent, a plant-derived agent, a biologic agent, an interleukin, an interferon, a cytokine, an inhibitor of histone deacetylase, a platinum compound, a taxane, an anti-neoplastic agent, an immune-modulating agent, or a monoclonal antibody.
10. The pharmaceutical combination of claim 1 , wherein the second therapeutic agent is a platinum compound.
11. The pharmaceutical combination of claim 10 , wherein the platinum compound is carboplatin.
12. The pharmaceutical combination of claim 1 , wherein the dinucleotide analogue is provided in a dosage form, wherein the dosage form comprises a pharmaceutically-acceptable carrier.
13. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier is a solution.
14. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier is an aqueous solution.
15. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier has less than 40% water.
16. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier has less than 5% water.
17. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier has less than 1% water.
18. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier is substantially anhydrous.
19. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier comprises a solvent selected from the group consisting of ethanol, glycerin, propylene glycol, polyethylene glycol, and combinations thereof.
20. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier is a solution comprising ethanol, glycerin, propylene glycol, polyethylene glycol, or a combination thereof.
21. The pharmaceutical combination of claim 12 , wherein the pharmaceutically-acceptable carrier comprises a concentration of propylene glycol and a concentration of glycerine.
22. The pharmaceutical combination of claim 21 , wherein the concentration of propylene glycol in the pharmaceutically-acceptable carrier is between 10-80%.
23. The pharmaceutical combination of claim 21 , wherein the concentration of propylene glycol in the pharmaceutically-acceptable carrier is between 50-70%.
24. The pharmaceutical combination of claim 12 , wherein the dosage form is formulated for administration by injection.
25. The pharmaceutical combination of claim 12 , wherein the dosage form is formulated for administration by subcutaneous injection.
26. The pharmaceutical combination of claim 12 , wherein the dosage form comprises between 0.1 and 200 mg of the dinucleotide analogue per mL of the pharmaceutically-acceptable carrier.
27. The pharmaceutical combination of claim 12 , wherein the dosage form comprises between 0.1 and 100 mg of the dinucleotide analogue per mL of the pharmaceutically-acceptable carrier.