1. An isolated polynucleotide encoding a polypeptide of interest, said isolated polynucleotide comprising:
(a) a sequence of n number of linked nucleosides or nucleotides comprising at least one modified nucleoside or nucleotide as compared to the chemical structure of an A, G, U or C nucleoside or nucleotide,
(b) a 5\u2032 UTR comprising at least one Kozak sequence,
(c) a 3\u2032 UTR, and
(d) at least one 5\u2032 cap structure.
2. The isolated polynucleotide of claim 1, further comprising a poly-A tail.
3. The isolated polynucleotide of claim 2 which is purified.
4. The isolated polynucleotide of claim 3, wherein the at least one 5\u2032 cap structure is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2\u2032fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.
5. The isolated polynucleotide of claim 4, wherein the modification of the nucleoside is located in the nucleoside base andor sugar portion of the nucleoside.
6. The isolated polynucleotide of claim 5, wherein the modification is located in the nucleotide base and the nucleoside base has the formula:
wherein:
denotes a single or double bond;
X is O or S;
U and W are each independently C or N;
V is O, S, C or N;
wherein when V is C then R1 is H, C1-6 alkyl, C1-6 alkenyl, C1-6 alkynyl, halo, or \u2014ORc,
wherein C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl are each optionally substituted with \u2014OH, \u2014NRaRb, \u2014SH, \u2014C(O)Rc, \u2014C(O)ORc, \u2014NHC(O)Rc, or \u2014NHC(O)ORc;
and wherein when V is O, S, or N then R1 is absent;
R2 is H, \u2014ORc, \u2014SRc, \u2014NRaRb, or halo;
or when V is C then R1 and R2 together with the carbon atoms to which they are attached can form a 5- or 6-membered ring optionally substituted with 1-4 substituents selected from halo, \u2014OH, \u2014SH, \u2014NRaRb, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, C1-20 alkoxy, or C1-20 thioalkyl;
R3 is H or C1-20 alkyl;
R4 is H or C1-20 alkyl; wherein when denotes a double bond then R4 is absent, or N\u2014R4, taken together, forms a positively charged N substituted with C1-20 alkyl;
Ra and Rb are each independently H, C1-20 alkyl, C2-20 alkenyl, C2-20 alkynyl, or C6-20 aryl; and
Rc is H, C1-20 alkyl, C2-20 alkenyl, phenyl, benzyl, a polyethylene glycol group, or an amino-polyethylene glycol group.
7. The isolated polynucleotide of claim 6, wherein the modification is located in the nucleoside base and the nucleoside base has the formula:
wherein:
R3 is C1-20 alkyl.
8. The isolated polynucleotide of claim 7, wherein R3 is C1-4 alkyl.
9. The isolated polynucleotide of claim 7, wherein R3 is CH3.
10. The isolated polynucleotide of claim 1, wherein the modified nucleoside is not pseudouridine (\u03c8) or 5-methyl-cytidine (m5C).
11. A pharmaceutical composition comprising the isolated polynucleotide of claim 6 and a pharmaceutically acceptable excipient.
12. The pharmaceutical composition of claim 11, wherein the excipient is selected from a solvent, aqueous solvent, non-aqueous solvent, dispersion media, diluent, dispersion, suspension aid, surface active agent, isotonic agent, thickening or emulsifying agent, preservative, lipid, lipidoids liposome, lipid nanoparticle, core-shell nanoparticles, polymer, lipoplexe peptide, protein, cell, hyaluronidase, and mixtures thereof.
13. A method of increasing the level of a polypeptide of interest in a mammalian subject comprising administering to said subject the isolated polynucleotide of claim 6.
14. The method of claim 13, wherein the polynucleotide is formulated.
15. The method of claim 13, wherein isolated polynucleotide has a Protein:Cytokine Ratio of greater than 100 for either TNF-alpha or IFN-alpha.
16. The method of claim 13, wherein the isolated polynucleotide is administered at a total daily dose of between 1 ug and 150 ug.
17. The method of claim 16, wherein administration is by injection.
18. The method of claim 16, wherein administration is intradermal or subcutaneous or intramuscular.
19. The method of claim 13, wherein levels of the polypeptide of interest in the serum of the mammal are at least 50 pgmL at least two hours after administration.
20. The method of claim 19, wherein the levels of the polypeptide of interest in the serum of the mammal remain above 50 pgmL for at least 72 hours after administration.
21. The method of claim 20, wherein the levels of the polypeptide of interest in the serum of the mammal remain above 60 pgmL for at least 72 hours after administration.
22. The method of claim 13, wherein administration is in two or more equal or unequal split doses.
23. The method of claim 22, wherein the level of the polypeptide produced by the subject by administering split doses of the polynucleotide is greater than the levels produced by administering the same total daily dose of polynucleotide as a single administration.
24. The method of claim 13, wherein the mammalian subject is a human patient in need of an increased level of the polypeptide of interest.
25. The method of claim 24, wherein the increased level of the polypeptide of interest is detectable in a bodily fluid of said patient.
26. The method of claim 53, wherein the bodily fluid is selected from the group consisting of peripheral blood, serum, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, amniotic fluid, cerumen, breast milk, broncheoalveolar lavage fluid, semen, prostatic fluid, cowper’s fluid or pre-ejaculatory fluid, sweat, fecal matter, hair, tears, cyst fluid, pleural and peritoneal fluid, pericardial fluid, lymph, chyme, chyle, bile, interstitial fluid, menses, pus, sebum, vomit, vaginal secretions, mucosal secretion, stool water, pancreatic juice, lavage fluids from sinus cavities, bronchopulmonary aspirates, blastocyl cavity fluid, and umbilical cord blood.
27. The method of claim 24, wherein administration is according to a dosing regimen which occurs over the course of hours, days, weeks, months, or years.
28. The method of claim 17, wherein injection is achieved by using one or more devices selected from multi-needle injection systems, catheter or lumen systems, and ultrasound, electrical or radiation based systems.
29. The method of claim 22, wherein the amount polynucleotide administered in any dose is substantially equal.
30. The method of claim 22, wherein a first dose, a second dose or any of a plurality of doses are administered at substantially the same time.
31. The method of claim 13, wherein administration comprises a single unit dose between about 10 mgkg and about 500 mgkg.
32. The method of claim 13, wherein administration comprises a single unit dose between about 1.0 mgkg and about 10 mgkg.
33. The method of claim 13, wherein administration comprises a single unit dose between about 0.001 mgkg and about 1.0 mgkg.
34. The isolated polynucleotide of claim 4, wherein the modification of the nucleoside includes replacing or substituting an atom of a pyrimidine nucleobase with an amine, an SH, a methyl or ethyl, or a chloro or fluoro.
35. The isolated polynucleotide of claim 1, wherein the n number of linked nucleosides comprise the Formula (Ia):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
U is O, S, N(RU)nu, or C(RU)nu, wherein nu is an integer from 0 to 2 and each RU is, independently, H, halo, or optionally substituted alkyl;
is a single or double bond;
– – – is a single bond or absent;
each of R1\u2032, R2\u2032, R1\u2033, R2\u2033, R1, R2, R3, R4, and R5, if present is, independently, H, halo hydroxy, thiol, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted hydroxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, or absent; wherein the combination of R3 with one or more of R1\u2032, R1\u2033, R2\u2032, R2\u2033, or R5 can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; wherein the combination of R5 with one or more of R1\u2032, R1\u2033, R2\u2032, or R2\u2033 can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; and wherein the combination of R4 and one or more of R1\u2032, R1\u2033, R2\u2032, R2\u2033, R3, or R5 can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl;
each of m\u2032 and m\u2033 is, independently, an integer from 0 to 3;
each of Y1, Y2, and Y3, is, independently, O, S, Se, \u2014NRN1\u2014, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, or absent;
each Y4 is, independently, H, hydroxy, thiol, boranyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, optionally substituted alkoxyalkoxy, or optionally substituted amino;
each Y5 is, independently, O, S, Se, optionally substituted alkylene, or optionally substituted heteroalkylene;
n is an integer from 1 to 100,000; and
B is a nucleobase.
36. The isolated polynucleotide of claim 35, wherein B is not pseudouridine (\u03c8) or 5-methyl-cytidine (m5C).
37. The isolated polynucleotide of claim 35 or 36, wherein:
U is O or C(RU)nu, wherein nu is an integer from 1 to 2 and each RU is, independently, H, halo, or optionally substituted alkyl;
each of R1, R1\u2032, R1\u2033, R2, R2\u2032, and R2\u2033, if present, is independently, H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl;
each of R3 and R4 is, independently, H, halo, hydroxy, optionally substituted alkyl, or optionally substituted alkoxyalkoxy;
each of Y1, Y2, and Y3, is, independently, O, S, Se, \u2014NRN1\u2014, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1 is H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl;
each Y4 is, independently, H, hydroxy, thiol, boranyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino;
each Y5 is, independently, O or optionally substituted alkylene; and
n is an integer from 10 to 10,000.
38. The isolated polynucleotide of claim 37, wherein each of R1, R1\u2033, and R1\u2033, if present, is H.
39. The isolated polynucleotide of claim 38, wherein each of R2, R2\u2032, and R2\u2033, if present, is, independently, H, halo, hydroxy, optionally substituted alkoxy, or optionally substituted alkoxyalkoxy.
40. The isolated polynucleotide of claim 39, wherein each of R2, R2\u2032, and R2\u2033, if present, is H.
41. The isolated polynucleotide of claim 40, wherein each of R1, R1\u2032, and R1\u2033, if present, is, independently, H, halo, hydroxy, optionally substituted alkoxy, or optionally substituted alkoxyalkoxy.
42. The isolated polynucleotide of claim 35, wherein the n number of linked nucleotides comprise the Formula (IIa):
or a pharmaceutically acceptable salt or stereoisomer thereof.
43. The isolated polynucleotide of claim 42, wherein the n number of linked nucleosides comprise the Formula (IIb) or (IIc):
or a pharmaceutically acceptable salt thereof.
44. The isolated polynucleotide of claim 43, wherein the n number of linked nucleosides comprise the Formula (IIb-1), (IIb-2), or (IIc-1)-(IIc-4).
45. The isolated polynucleotide of claim 35, wherein the n number of linked nucleosides comprise the Formula (IId):
or a pharmaceutically acceptable salt or stereoisomer thereof.
46. The isolated polynucleotide of claim 44, wherein the n number of linked nucleosides comprise the Formula (IIe) or (IIf):
or a pharmaceutically acceptable salt thereof.
47. The isolated polynucleotide of claim 35, wherein each of said linked nucleotides independently have one of Formulas (IIg)-(IIj):
or a pharmaceutically acceptable salt or stereoisomer thereof.
48. The isolated polynucleotide of claim 35, wherein the n number of linked nucleosides comprise the Formula (IIk):
or a pharmaceutically acceptable salt or stereoisomer thereof.
49. The isolated polynucleotide of claim 48, wherein the n number of linked nucleosides comprise the Formula (IIl):
or a pharmaceutically acceptable salt or stereoisomer thereof.
50. The isolated polynucleotide of claim 48, wherein the n number of linked nucleosides comprise the Formula (IIm):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
each of R1\u2032, R1\u2033, R2\u2032, and R2\u2033 is, independently, H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, or absent; and wherein the combination of R2\u2032 and R3 or the combination of R2\u2033 and R3 can be taken together to form optionally substituted alkylene or optionally substituted heteroalkylene.
51. The isolated polynucleotide of any one of claims 38-50, wherein
U is O or C(RU)nu, wherein nu is an integer from 1 to 2 and each RU is, independently, H, halo, or optionally substituted alkyl;
each of R1 and R2 is, independently, H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl;
each of R3 and R4 is, independently, H or optionally substituted alkyl;
each of Y1, Y2, and Y3, is, independently, O, S, Se, \u2014NRN1\u2014, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1 is H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl;
each Y4 is, independently, H, hydroxy, thiol, boranyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino;
each Y5 is, independently, O or optionally substituted alkylene; and
n is an integer from 10 to 10,000.
52. The isolated polynucleotide of claim 35, wherein the n number of linked nucleosides comprise the Formula (IIn):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
U is O or C(RU)nu, wherein nu is an integer from 1 to 2 and each RU is, independently, H, halo, or optionally substituted alkyl;
each of R1 and R4 is, independently, H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl;
R3\u2032 is O, S, or \u2014NRN1\u2014, wherein RN1 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, or optionally substituted aryl;
R3\u2033 is optionally substituted alkylene or optionally substituted heteroalkylene;
each of Y1, Y2, and Y3, is, independently, O, S, Se, \u2014NRN1\u2014, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1 is H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl;
each Y4 is, independently, H, hydroxy, thiol, boranyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino;
each Y5 is, independently, O, S, optionally substituted alkylene (e.g., methylene), or optionally substituted heteroalkylene; and
n is an integer from 10 to 10,000.
53. The isolated polynucleotide of claim 35, wherein the n number of linked nucleosides comprise Formula (IIn-1) or (II-n2).
54. The isolated polynucleotide of any one of claims 35-53, wherein each B independently has a formula selected from Formula (b1)-(b5):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
is a single or double bond;
each of T\u2033, T1\u2033, T2\u2032, and T2\u2033 is, independently, H, optionally substituted alkyl, optionally substituted alkoxy, or optionally substituted thioalkoxy, or the combination of T1\u2032 and T1\u2033 or the combination of T2\u2032 and T2\u2033 join together (e.g., as in T2) to form O (oxo), S (thio), or Se (seleno);
each of V1 and V2 is, independently, O, S, N(RVb)nv, or C(RVb)nv, wherein nv is an integer from 0 to 2 and each RVb is, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted acylaminoalkyl, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, or optionally substituted alkoxycarbonylalkoxy;
R10 is H, halo, optionally substituted amino acid, hydroxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aminoalkyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted alkoxy, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted carboxyalkoxy, optionally substituted carboxyalkyl, or optionally substituted carbamoylalkyl;
R11 is H or optionally substituted alkyl;
R12a is H, optionally substituted alkyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl, optionally substituted carboxyalkyl, optionally substituted carboxyalkoxy, optionally substituted carboxyaminoalkyl, or optionally substituted carbamoylalkyl; and
R12c is H, halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted thioalkoxy, optionally substituted amino, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl.
55. The isolated polynucleotide of any one of claims 35-54, wherein B comprises Formula (b6)-(b9):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
is a single or double bond;
each of T1\u2032, T1\u2033, T2\u2032, and T2\u2033 is, independently, H, optionally substituted alkyl, optionally substituted alkoxy, or optionally substituted thioalkoxy, or the combination of T1\u2032 and T1\u2033 join together or the combination of T2\u2032 and T2\u2033 join together to form O (oxo), S (thio), or Se (seleno), or each T1 and T2 is, independently, O (oxo), S (thio), or Se (seleno);
each of W1 and W2 is, independently, N(RWa)nw or C(RWa)nw, wherein nw is an integer from 0 to 2 and each RWa is, independently, H, optionally substituted alkyl, or optionally substituted alkoxy;
each V3 is, independently, O, S, N(RVa)nv, or C(RVa)nv, wherein nv is an integer from 0 to 2 and each RVa is, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, optionally substituted alkoxy, optionally substituted alkenyloxy, or optionally substituted alkynyloxy, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted acylaminoalkyl, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, optionally substituted alkoxycarbonylacyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted carboxyalkyl, optionally substituted carboxyalkoxy, optionally substituted carboxyaminoalkyl, or optionally substituted carbamoylalkyl, and wherein RVa and R12c taken together with the carbon atoms to which they are attached can form optionally substituted cycloalkyl, optionally substituted aryl, or optionally substituted heterocyclyl;
R12a is H, optionally substituted alkyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted carboxyalkyl, optionally substituted carboxyalkoxy, optionally substituted carboxyaminoalkyl, optionally substituted carbamoylalkyl, or absent;
R12b is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted alkaryl, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, optionally substituted amino acid, optionally substituted alkoxycarbonylacyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted carboxyalkyl, optionally substituted carboxyalkoxy, optionally substituted carboxyaminoalkyl, or optionally substituted carbamoylalkyl,
wherein the combination of R12b and T1\u2032 or the combination of R12b and R12c can join together to form optionally substituted heterocyclyl; and
R12c is H, halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted thioalkoxy, optionally substituted amino, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl.
56. The isolated polynucleotide of claim 55, wherein R12a, R12b, R12c, or RVa is substituted with \u2014(CH2)s2(OCH2CH2)s1(CH2)s3OR\u2032, wherein s1 is an integer from 1 to 10, each of s2 and s3, independently, is an integer from 0 to 10, and R\u2032 is H or C1-20 alkyl); or \u2014NRN1(CH2)s2(CH2CH2O)s1(CH2)s3NRN1, wherein s1 is an integer from 1 to 10, each of s2 and s3, independently, is an integer from 0 to 10, and each RN1 is, independently, hydrogen or optionally substituted C1-6 alkyl.
57. The isolated polynucleotide of claim 55, wherein B comprises Formula (b28)-(b31).
58. The isolated polynucleotide of any one of claims 35-57, wherein B comprises Formula (b10)-(b14):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
each of T3\u2032 and T3\u2033 is, independently, H, optionally substituted alkyl, optionally substituted alkoxy, or optionally substituted thioalkoxy, or the combination of T3\u2032 and T3\u2033 join together to form O (oxo), S (thio), or Se (seleno);
each V4 is, independently, O, S, N(RVc)nv, or C(RVc)nv, wherein nv is an integer from 0 to 2 and each RVc is, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, or optionally substituted alkynyloxy, wherein the combination of R13b and RVc can be taken together to form optionally substituted heterocyclyl;
each V5 is, independently, N(RVd)nv, or C(RVd)nv, wherein nv is an integer from 0 to 2 and each RVd is, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, or optionally substituted alkynyloxy;
each of R13a and R13b is, independently, H, optionally substituted acyl, optionally substituted acyloxyalkyl, optionally substituted alkyl, or optionally substituted alkoxy, wherein the combination of R13b and R14 can be taken together to form optionally substituted heterocyclyl;
each R14 is, independently, H, halo, hydroxy, thiol, optionally substituted acyl, optionally substituted amino acid, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted acyloxyalkyl, optionally substituted amino, azido, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, or optionally substituted aminoalkynyl; and
each of R15 and R16 is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl.
59. The isolated polynucleotide of claim 5858, wherein B comprises Formula (b32)-(b36).
60. The isolated polynucleotide of any one of claims 35-59, wherein B comprises Formula (b15)-(b17):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein
each of T4\u2032, T4\u2033, T5\u2032, T5\u2033, T6\u2032, and T6\u2033 is independently, H, optionally substituted alkyl, or optionally substituted alkoxy, and wherein the combination of T4\u2032 and T4\u2033 or the combination of T5\u2032 and T5\u2033 or the combination of T6\u2032 and T6\u2033 join together form O (oxo), S (thio), or Se (seleno);
each of V5 and V6 is, independently, O, S, N(RVd)nv, or C(RVd)nv, wherein nv is an integer from 0 to 2 and each RVd is, independently, H, halo, thiol, optionally substituted amino acid, cyano, amidine, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino; and
each of R17, R18, R19a, R19b, R21, R22, R23, and R24 is, independently, H, halo, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted thioalkoxy, optionally substituted amino, or optionally substituted amino acid.
61. The isolated polynucleotide of claim 60, wherein B comprises Formula (b37)-(b40).
62. The isolated polynucleotide of any one of claims 35-59, wherein B comprises Formula (b18)-(b20):
or a pharmaceutically acceptable salt or stereoisomer thereof
wherein
each V7 is, independently, O, S, N(RVe)nv, or C(RVe)nv, wherein nv is an integer from 0 to 2 and each RVe is, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, or optionally substituted alkynyloxy;
each R25 is, independently, H, halo, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted thioalkoxy, or optionally substituted amino;
each of R26a and R26b is, independently, H, optionally substituted acyl, optionally substituted amino acid, optionally substituted carbamoylalkyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted hydroxyalkynyl, optionally substituted alkoxy, or polyethylene glycol group, or an amino-polyethylene glycol group;
each R27 is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted thioalkoxy, or optionally substituted amino;
each R28 is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; and
each R29 is, independently, H, optionally substituted acyl, optionally substituted amino acid, optionally substituted carbamoylalkyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted alkoxy, or optionally substituted amino.
63. The isolated polynucleotide of claim 62, wherein B comprises Formula (b41)-(b43).
64. The isolated polynucleotide of claim 62, wherein R26a, R26b, or R29 is substituted with \u2014(CH2)s2(OCH2CH2)s1(CH2)s3OR\u2032, wherein s1 is an integer from 1 to 10, each of s2 and s3, independently, is an integer from 0 to 10, and R\u2032 is H or C1-20 alkyl); or \u2014NRN1(CH2)s2(CH2CH2O)s1(CH2)s3NRN1, wherein s1 is an integer from 1 to 10, each of s2 and s3, independently, is an integer from 0 to 10, and each RN1 is, independently, hydrogen or optionally substituted C1-6 alkyl.
65. The isolated polynucleotide of any one of claims 35-64, wherein B comprises Formula (b21):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein X12 is, independently, O, S, optionally substituted alkylene, or optionally substituted heteroalkylene; xa is an integer from 0 to 3; R12a is H, optionally substituted alkyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, or absent; and T2 is O, S, or Se.
66. The isolated polynucleotide of any one of claims 35-65, wherein B comprises Formula (b22):
or a pharmaceutically acceptable salt or stereoisomer thereof,
wherein R10\u2032 is independently, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, optionally substituted alkoxy, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted carboxyalkoxy, optionally substituted carboxyalkyl, or optionally substituted carbamoylalkyl; R11 is H or optionally substituted alkyl; R12a is H, optionally substituted alkyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, or absent; and each of T1 and T2 is, independently, O, S, or Se.
67. The isolated polynucleotide of any one of claims 35-66, wherein B comprises Formula (b23):
wherein R10 is optionally substituted heterocyclyl or optionally substituted aryl; R11 is H or optionally substituted alkyl; R12a is H, optionally substituted alkyl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynyl, or absent; and each of T1 and T2 is, independently, O, S, or Se.
68. The isolated polynucleotide of any one of claims 35-67, wherein B comprises Formula (b24) or (b25):
wherein
T3 is O, S, or Se;
each of R13a and R13b is, independently, H, optionally substituted acyl, optionally substituted alkyl, or optionally substituted alkoxy, wherein the combination of R13b and R14 can be taken together to form optionally substituted heterocyclyl;
R14\u2032 is independently, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkheterocyclyl, optionally substituted alkaryl, optionally substituted aminoalkyl, optionally substituted aminoalkenyl, optionally substituted aminoalkynynyl, optionally substituted alkoxy, optionally substituted alkoxycarbonylalkyl, optionally substituted alkoxycarbonylalkenyl, optionally substituted alkoxycarbonylalkynyl, optionally substituted alkoxycarbonylalkoxy, optionally substituted carboxyalkoxy, optionally substituted carboxyalkyl, or optionally substituted carbamoylalkyl; and
each R15 is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl.
69. The isolated polynucleotide of any one of claims 35-68, wherein B comprises Formula (b26) or (b27):
70. The polynucleotide of any one of claims 35-69, wherein B comprises Formula (b28)-b43).
71. The isolated polynucleotide of claim 35, wherein said isolated polynucleotide is prepared from one or more building blocks selected from BB-1 to BB-274, or a pharmaceutically acceptable salt or stereoisomer thereof.
72. The isolated polynucleotide of claim 35, wherein said isolated polynucleotide is prepared from one or more building blocks selected from compound 1-50, or a pharmaceutically acceptable salt or stereoisomer thereof.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A system comprising:
a primary inertial measurement unit configured to generate a primary position signal, the primary position signal indicative of an estimated position of a machine on a worksite;
a secondary inertial measurement unit configured to generate a secondary position signal, the secondary position signal indicative of the estimated position of the machine; and
an error detection module communicably coupled to the primary inertial measurement unit and the secondary inertial measurement unit, the error detection module configured to:
receive the primary position signal from the primary inertial measurement unit;
receive the secondary position signal from the secondary inertial measurement unit;
detect if an out-range error is present in at least one of the primary position signal and the secondary position signal, wherein the presence of the out-range error is based on the respective primary position signal or the secondary position signal exceeding a first predetermined threshold;
detect if an in-range error is present in at least one of the primary position signal and the secondary position signal based, at least in part, on the presence of the out-range error in the respective primary position signal or the secondary position signal, wherein the presence of the in-range error is based on the respective primary position signal or the secondary position signal exceeding a second predetermined threshold; and
determine an action to be performed based, at least in part, on the presence of at least one of the out-range error and the in-range error in the respective primary position signal or the secondary position signal.
2. The system of claim 1, wherein the first predetermined threshold is at least one of:
a threshold gyroscopic value; and
a threshold acceleration value.
3. The system of claim 1, wherein the second predetermined threshold is based on a correlation between the primary position signal and the secondary position signal.
4. The system of claim 1, wherein the error detection module is communicably coupled to a transmission module associated with the machine.
5. The system of claim 4, wherein if the presence of the out-range error is detected in the primary position signal, the action includes any one of:
stopping the machine within at least one of a first predetermined distance and a first predetermined time duration; and
stopping the machine within at least one of a second predetermined distance and a second predetermined time duration, such that the first predetermined distance is greater than the second predetermined distance and the first predetermined time duration is greater than the second predetermined time duration.
6. The system of claim 5, wherein the machine is stopped within at least one of the first predetermined distance and the first predetermined time duration if the presence of the out-range error is not detected in the secondary position signal.
7. The system of claim 5, wherein the machine is stopped within at least one of the second predetermined distance and the second predetermined time duration if the presence of the out-range error is detected in the secondary position signal.
8. The system of claim 1, wherein the error detection module is communicably coupled to a position detection system associated with the machine, the position detection system configured to generate a signal indicative of a current position of the machine on the worksite.
9. The system of claim 8, wherein if the presence of the out-range error is not detected in the primary position signal and if the presence of any of the out-range error and the in-range error is detected in the secondary position signal, the action includes:
adding a predetermined noise to the primary position signal; and
forwarding the primary position signal with the added predetermined noise to the position detection system.
10. The system of claim 8, wherein if the presence of the in-range error is not detected in the primary position signal and the secondary position signal, the action includes:
forwarding the primary position signal to the position detection system.
11. A method comprising:
receiving a primary position signal from a primary inertial measurement unit, wherein the primary position signal is indicative of an estimated position of a machine on a worksite;
receiving a secondary position signal from a secondary inertial measurement unit, wherein the secondary position signal is indicative of the estimated position of the machine;
detecting if an out-range error is present in at least one of the primary position signal and the secondary position signal, wherein the presence of the out-range error is based on the respective primary position signal or the secondary position signal exceeding a first predetermined threshold;
detecting if an in-range error is present in at least one of the primary position signal and the secondary position signal based, at least in part, on the presence of the out-range error in the respective primary position signal or the secondary position signal, wherein the presence of the in-range error is based on the respective primary position signal or the secondary position signal exceeding a second predetermined threshold; and
determining an action to be performed based, at least in part, on the presence of at least one of the out-range error and the in-range error in the respective primary position signal or the secondary position signal.
12. The method of claim 11, wherein the first predetermined threshold is at least one of:
a threshold gyroscopic value; and
a threshold acceleration value.
13. The method of claim 11, wherein the second predetermined threshold is based on a correlation between the primary position signal and the secondary position signal.
14. The method of claim 11, wherein if the presence of the out-range error is detected in the primary position signal, the action includes any one of:
stopping the machine within at least one of a first predetermined distance and a first predetermined time duration; and
stopping the machine within at least one of a second predetermined distance and a second predetermined time duration, such that the first predetermined distance is greater than the second predetermined distance and the first predetermined time duration is greater than the second predetermined time duration.
15. The method of claim 14, wherein the machine is stopped within at least one of the first predetermined distance and the first predetermined time duration if the presence of the out-range error is not detected in the secondary position signal.
16. The method of claim 14, wherein the machine is stopped within at least one of the second predetermined distance and the second predetermined time duration if the presence of the out-range error is detected in the secondary position signal.
17. The method of claim 11, wherein if the presence of the out-range error is not detected in the primary position signal and if the presence of any of the out-range error and the in-range error is detected in the secondary position signal, the action includes:
adding a predetermined noise to the primary position signal; and
forwarding the primary position signal with the added predetermined noise to a position detection system.
18. The method of claim 11, wherein if the presence of the in-range error is not detected in the primary position signal and the secondary position signal, the action includes:
forwarding the primary position signal to a position detection system.
19. A machine comprising:
an engine;
a transmission module operatively coupled to the engine;
a position detection system, the position detection system configured to generate a signal indicative of a current position of the machine on a worksite;
a primary inertial measurement unit configured to generate a primary position signal, the primary position signal indicative of an estimated position of the machine on the worksite;
a secondary inertial measurement unit configured to generate a secondary position signal, the secondary position signal indicative of the estimated position of the machine; and
an error detection module communicably coupled to the primary inertial measurement unit and the secondary inertial measurement unit, the error detection module configured to:
receive the primary position signal from the primary inertial measurement unit;
receive the secondary position signal from the secondary inertial measurement unit;
detect if an out-range error is present in at least one of the primary position signal and the secondary position signal, wherein the presence of the out-range error is based on the respective primary position signal or the secondary position signal exceeding a first predetermined threshold;
detect if an in-range error is present in at least one of the primary position signal and the secondary position signal based, at least in part, on the presence of the out-range error in the respective primary position signal or the secondary position signal, wherein the presence of the in-range error is based on the respective primary position signal or the secondary position signal exceeding a second predetermined threshold; and
determine an action to be performed based, at least in part, on the presence of at least one of the out-range error and the in-range error in the respective primary position signal or the secondary position signal.
20. The machine of claim 19, wherein the error detection module is further communicably coupled to the position detection system and the transmission module.