U.S. patent application number 16/692966 was filed with the patent office on 2020-06-25 for methods for the preparation of ribosides.
The applicant listed for this patent is Gilead Sciences, Inc.. Invention is credited to Steven Donald Axt, Pavel Robertovich Badalov, Katrien Brak, Silvio Campagna, Andrei Chtchemelinine, Edward Doerffler, Morin Mae Frick, Detian Gao, Lars V. Heumann, Brittanie Hoang, Willard Lew, Robert Ronald Milburn, Sean Timothy Neville, Bruce Ross, Erik Rueden, Robert William Scott, Dustin Siegel, Andrew C. Stevens, Clarissa Tadeus, Tiago Vieira, Andrew W. Waltman, Xianghong Wang, Mark Charles Whitcomb, Lydia Wolfe, Chia-Yun Yu.
Application Number | 20200197422 16/692966 |
Document ID | / |
Family ID | 54557474 |
Filed Date | 2020-06-25 |
View All Diagrams
United States Patent
Application |
20200197422 |
Kind Code |
A1 |
Axt; Steven Donald ; et
al. |
June 25, 2020 |
METHODS FOR THE PREPARATION OF RIBOSIDES
Abstract
Provided are methods of preparing compounds and pharmaceutical
compositions for treating Filoviridae virus infections The
compounds, compositions, and methods provided are particularly
useful for the treatment of Marburg virus, Ebola virus and Cueva
virus infections.
Inventors: |
Axt; Steven Donald; (San
Mateo, CA) ; Badalov; Pavel Robertovich; (Edmonton,
CA) ; Brak; Katrien; (Belmont, CA) ; Campagna;
Silvio; (Dover, DE) ; Chtchemelinine; Andrei;
(San Mateo, CA) ; Doerffler; Edward; (Union City,
CA) ; Frick; Morin Mae; (San Mateo, CA) ; Gao;
Detian; (Edmonton, CA) ; Heumann; Lars V.;
(Redwood City, CA) ; Hoang; Brittanie; (Berkeley,
CA) ; Lew; Willard; (San Mateo, CA) ; Milburn;
Robert Ronald; (San Mateo, CA) ; Neville; Sean
Timothy; (San Mateo, CA) ; Ross; Bruce; (El
Granada, CA) ; Rueden; Erik; (San Mateo, CA) ;
Scott; Robert William; (San Mateo, CA) ; Siegel;
Dustin; (Half Moon Bay, CA) ; Stevens; Andrew C.;
(Edmonton, CA) ; Tadeus; Clarissa; (San Francisco,
CA) ; Vieira; Tiago; (Edmonton, CA) ; Waltman;
Andrew W.; (Kalamazoo, MI) ; Wang; Xianghong;
(Dublin, CA) ; Whitcomb; Mark Charles; (Woodside,
CA) ; Wolfe; Lydia; (San Mateo, CA) ; Yu;
Chia-Yun; (Burlingame, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Sciences, Inc. |
Foster City |
CA |
US |
|
|
Family ID: |
54557474 |
Appl. No.: |
16/692966 |
Filed: |
November 22, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14926063 |
Oct 29, 2015 |
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16692966 |
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62105619 |
Jan 20, 2015 |
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62072331 |
Oct 29, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/7056 20130101;
A61K 31/665 20130101; C07H 15/18 20130101; C07D 519/00 20130101;
A61K 31/683 20130101; A61P 31/12 20180101; A61K 45/06 20130101;
C07F 9/2429 20130101; C07H 1/00 20130101; A61K 31/706 20130101;
A61K 31/685 20130101; A61P 31/14 20180101; C07F 9/65616 20130101;
A61K 31/53 20130101; A61K 31/675 20130101; C07D 487/04 20130101;
C07H 11/00 20130101; A61K 31/6615 20130101; A61K 31/00 20130101;
A61P 31/00 20180101; C07H 1/02 20130101; A61P 43/00 20180101; A61K
31/53 20130101; A61K 2300/00 20130101; A61K 31/675 20130101; A61K
2300/00 20130101; A61K 31/685 20130101; A61K 2300/00 20130101; A61K
31/706 20130101; A61K 2300/00 20130101; A61K 31/7056 20130101; A61K
2300/00 20130101 |
International
Class: |
A61K 31/685 20060101
A61K031/685; A61K 31/706 20060101 A61K031/706; A61K 31/7056
20060101 A61K031/7056; A61K 31/53 20060101 A61K031/53; C07H 1/00
20060101 C07H001/00; C07H 1/02 20060101 C07H001/02; C07H 11/00
20060101 C07H011/00; C07H 15/18 20060101 C07H015/18; A61K 31/6615
20060101 A61K031/6615; A61K 31/665 20060101 A61K031/665; A61K
31/683 20060101 A61K031/683; C07D 487/04 20060101 C07D487/04; C07D
519/00 20060101 C07D519/00; C07F 9/24 20060101 C07F009/24; C07F
9/6561 20060101 C07F009/6561; A61K 31/675 20060101 A61K031/675;
A61K 45/06 20060101 A61K045/06; A61K 31/00 20060101 A61K031/00 |
Claims
1.-42. (canceled)
43. A method of preparing a compound of Formula (VIII) having
structure: ##STR00210## the method comprising: (a) (i) forming a
first reaction mixture having BCl.sub.3, dichloromethane, and a
compound of Formula (XI-a): ##STR00211## and (ii) combining
K.sub.2CO.sub.3 with the first reaction mixture to prepare the
compound of Formula (XI-b): ##STR00212## (b) forming a second
reaction mixture having acetone, 2,2-dimethoxypropane, sulfuric
acid, and the compound of Formula (XI-b) to prepare a compound of
Formula (XI-c): ##STR00213## and (c) forming a third reaction
mixture having: (i) MgCl.sub.2, (ii) diisopropylethylamine, (iii)
the compound of Formula (XI-c), and (iv) a compound of Formula (X)
having structure: ##STR00214## to prepare the compound of Formula
(VIII).
44. The method of claim 43, further comprising preparing the
compound of Formula (XI-a) by a method comprising: forming a
reaction mixture with a cyanating agent, a Lewis acid, a Bronsted
acid, a solvent, and a compound of Formula (V-a) or Formula (V-b)
having structure: ##STR00215## to prepare the compound of Formula
(XI-a).
45. The method of claim 44, wherein the cyanating agent is TMSCN,
TBSCN, TESCN, HCN, KCN, NaCN, 4-toluenesulfonyl cyanide, CuCN,
CuCN*LiCl, LiCN, Zn(CN).sub.2, K.sub.4 [Fe(CN).sub.6],
tetrabutylammonium cyanide, tetramethylammonium cyanide,
tetraethylammonium cyanide, tetrabutylammonium cyanide,
tetraalkylammonium cyanide with alkyl independently being Me, Et,
Pr, iPr, Bu, iBu, tertBu, Pent, Hex, tributyltin cyanide,
trimethyltin cyanide, triethyltin cyanide, tripropyltin cyanide,
trialkyltin cyanide with alkyl independently being Me, Et, Pr, iPr,
Bu, iBu, tertBu, Pent, Hex, 2-hydroxy-2-methylpropanenitrile; or
combinations thereof.
46. The method of claim 44, wherein the Lewis acid is TMSOTf,
TMSOTf, TBSOTf, TESOTf, BF.sub.3, BF.sub.3-OEt.sub.2, BCl.sub.3,
BF.sub.3-THF, MgCl.sub.2, MgI.sub.2, MgBr.sub.2,
MgBr.sub.2-OEt.sub.2, ZnCl.sub.2, ZnBr.sub.2, ZnI.sub.2, LiCl,
LiBr, LiI, AlCl.sub.3, AlBr.sub.3, AlI.sub.3,
Me.sub.2Si(OTf).sub.2, Et.sub.2Si(OTf).sub.2,
Pr.sub.2Si(OTf).sub.2, iPr.sub.2Si(OTf).sub.2,
(tBu).sub.2Si(OTf).sub.2, (C.sub.6F.sub.5).sub.3B, MeSiCl.sub.3,
Me.sub.2SiCl.sub.2, SiCl.sub.4, TMSCl, TMSI, TMSVr, TBSCl, TBSBr,
TBSI, TESCl, TESBr, TESI, SmCl.sub.3, SmBr.sub.3, SmI.sub.2,
SmI.sub.3, ScI.sub.3, ScBr.sub.3, ScI.sub.3, Sm(OTf).sub.3,
Sc(OTf).sub.3, TiCl.sub.4, Ti(OiPr).sub.4, Ti(OiPr).sub.3Cl,
Ti(OiPr).sub.2Cl.sub.2, Ti(OiPr)Cl.sub.3, Zn(BF.sub.4).sub.2,
LiBF.sub.4, Mg(BF.sub.4).sub.2, ZrCl.sub.4, FeCl.sub.2, FeCl.sub.3,
FeBr.sub.2, FeBr.sub.3, FeI.sub.2, FeI.sub.3, Cu(OTf),
Cu(OTf).sub.2, 4-toluenesulfonylchloride, benzenesulfonylchloride,
4-toluenesulfonyl triflate, benzenesulfonyl triflate, methyl
sulfonyl chloride, methylsulfonic anhydrate, InCl.sub.3,
InBr.sub.3, InI.sub.3, In(OTf).sub.3, Mg(SO.sub.4).sub.2,
NaSO.sub.4; or combinations thereof.
47. The method of claim 44, wherein the Bronsted acid is TFA,
benzenesulfonic acid, HCl, 4-toluenesulfonic acid, triflic acid,
trifluoroacetic acid, 4-nitrobenzoic acid, methylsoulfonic acid,
sulfuric acid, phosphoric acid, HBr, acetic acid, formic acid, HI,
trifluoromethylsulfonic acid, 4-fluorobenzoic acid, pivalic acid,
HBF.sub.4, nitric acid, 4-chloro-benzoic acid, pentafluorophenol,
HPF.sub.6, Camphorsulfonic acid; or combinations thereof.
48. The method of claim 44, wherein the solvent is DCM, THF, MeTHF,
Et.sub.2O, MeCN, EtCN, toluene, benzene, chlorobenzene,
nitrobenzene, fluorobenzene, methanol, ethanol, 2-propanol,
propanol, butanol, MTBE, EtOAc, iPrOAc, Me.sub.2O, (TMS).sub.2O,
acetone, 2-butanone, chloroform, 1,2-dichloroethane, diglyme,
dioxane, acetic acid, formic acid, trifluoroacetic acid,
methylisobutylketone, DMAc, DMF, NMP, DMSO; or combinations
thereof.
49. The method of claim 44, further comprising preparing the
compound of Formula (V-a) or Formula (V-b) by a method comprising:
forming the reaction mixture with TMSCl, PhMgCl, iPrMgCl, an
additive, a compound of Formula (VI-a) having structure:
##STR00216## and a compound of Formula (VII): ##STR00217## to
prepare the compound of Formula (V-a) or Formula (V-b), wherein the
additive is BF.sub.3-OEt.sub.2, Sm(OTf).sub.3, Sc(OTf).sub.3,
FeCl.sub.3, LiCl, LiBr, TiCl(OiPr).sub.3, ScCl.sub.3,
Bu.sub.4NBr+LaCl.sub.3-2LiCl, nLaCl.sub.3+mLiCl, wherein m is 0.5
to 50, n is 1 to 100, LaCl.sub.3+2LiCl, Sm(OTf).sub.3+LiCl,
SmCl.sub.3, Bis[2-(N,N-dimethylamino)ethyl] ether, TMEDA,
NdCl.sub.3, NdCl.sub.3+CsCl, nNdCl.sub.3+mLiCl, wherein m is 0.5 to
50, n is 1 to 100, NdCl.sub.3+2LiCl, NdCl.sub.3+LiBr,
NdCl.sub.3+LiI, NdBr.sub.3, NdBr.sub.3+CsCl, nNdBr.sub.3+mLiCl,
wherein m is 0.5 to 50, n is 1 to 100, NdBr.sub.3+2LiCl,
NdBr.sub.3+LiBr, NdBr.sub.3+LiI, Nd(OTf).sub.3, CeCl.sub.3,
CeCl.sub.3+CsCl, nCeCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1
to 100, CeCl.sub.3+2LiCl, CeCl.sub.3+LiBr, CeCl.sub.3+LiI,
CeBr.sub.3, Ce(OTf).sub.3, YCl.sub.3, YCl.sub.3+CsCl,
nYCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to 100,
YCl.sub.3+2LiCl, YCl.sub.3+LiBr, YCl.sub.3+LiI, YBr.sub.3,
YBr.sub.3+CsCl, nYBr.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to
100, YBr.sub.3+2LiCl, YBr.sub.3+LiBr, YBr.sub.3+LiI, Y(OTf).sub.3,
LaCl.sub.3, La(OTf).sub.3, MgCl.sub.2, TiCl.sub.4, SnCl.sub.4,
AlCl.sub.3, Bu.sub.4NCl, Diethyleneglycol diethylether (DGDE),
DGDE+Bu.sub.4NCl, DGDE+Bu.sub.4NBr, DGDE+Bu.sub.4NI, CaCl.sub.2,
CaBr.sub.2, CaI.sub.2, Ca(OTf).sub.2, YCl.sub.3, YCl.sub.3-2LiCl,
YCl.sub.3--LiCl or a combination thereof.
50. The method of claim 49, wherein the additive is
LaCl.sub.3-2LiCl, YCl.sub.3, CeCl.sub.3, NdCl.sub.3, or
LaCl.sub.3.
51. A method of preparing a compound of Formula V-a or V-b:
##STR00218## the method comprising: forming a reaction mixture
comprising a deprotonating agent, a silylating agent, a coupling
agent, an additive, a compound of Formula VI-a: ##STR00219## and a
compound of Formula VII: ##STR00220## to prepare the compound of
Formula V-a or V-b, wherein each R.sup.b is independently benzyl
(Bn) or tert-butyldimethylsilyl (TBS); alternatively, two R.sup.b
groups on adjacent carbons can be combined to form a
--C(Me).sub.2-- group; and R.sup.10 is H or a silyl group; wherein
the additive is BF.sub.3-OEt.sub.2, Sm(OTf).sub.3, Sc(OTf).sub.3,
FeCl.sub.3, LiCl, LiBr, TiCl(OiPr).sub.3, ScCl.sub.3,
Bu.sub.4NBr+LaCl.sub.3-2LiCl, nLaCl.sub.3+mLiCl, wherein m is 0.5
to 50, n is 1 to 100, LaCl.sub.3+2LiCl, Sm(OTf).sub.3+LiCl,
SmCl.sub.3, Bis[2-(N,N-dimethylamino)ethyl] ether, TMEDA,
NdCl.sub.3, NdCl.sub.3+CsCl, nNdCl.sub.3+mLiCl, wherein m is 0.5 to
50, n is 1 to 100, NdCl.sub.3+2LiCl, NdCl.sub.3+LiBr,
NdCl.sub.3+LiI, NdBr.sub.3, NdBr.sub.3+CsCl, nNdBr.sub.3+mLiCl,
wherein m is 0.5 to 50, n is 1 to 100, NdBr.sub.3+2LiCl,
NdBr.sub.3+LiBr, NdBr.sub.3+LiI, Nd(OTf).sub.3, CeCl.sub.3,
CeCl.sub.3+CsCl, nCeCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1
to 100, CeCl.sub.3+2LiCl, CeCl.sub.3+LiBr, CeCl.sub.3+LiI,
CeBr.sub.3, Ce(OTf).sub.3, YCl.sub.3, YCl.sub.3+CsCl,
nYCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to 100,
YCl.sub.3+2LiCl, YCl.sub.3+LiBr, YCl.sub.3+LiI, YBr.sub.3,
YBr.sub.3+CsCl, nYBr.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to
100, YBr.sub.3+2LiCl, YBr.sub.3+LiBr, YBr.sub.3+LiI, Y(OTf).sub.3,
LaCl.sub.3, La(OTf).sub.3, MgCl.sub.2, TiCl.sub.4, SnCl.sub.4,
AlCl.sub.3, Bu.sub.4NCl, Diethyleneglycol diethylether (DGDE),
DGDE+Bu.sub.4NCl, DGDE+Bu.sub.4NBr, DGDE+Bu.sub.4NI, CaCl.sub.2,
CaBr.sub.2, CaI.sub.2, Ca(OTf).sub.2, YCl.sub.3, YCl.sub.3-2LiCl,
YCl.sub.3--LiCl or a combination thereof.
52. The method of claim 51, wherein the deprotonating agent is
sodium hydride (NaH), isopropylmagnesium chloride (iPrMgCl),
tert-butylmagnesium chloride (tBuMgCl), phenylmagnesium chloride
(PhMgCl), phenylmagnesium bromide (PhMgBr), butyllithium (BuLi),
methyllithium (MeLi), methylmagnesium chloride (MeMgCl),
methylmagnesium bromide (MeMgBr), tert-butyllithium (tBuLi),
isopropyllithium (iPrLi), phenyllithium (PhLi), lithium hydride
(LiH), potassium hydride (KH), ethyllithium (EtLi), ethylmagnesium
bromide (EtMgBr), ethylmagnesium chloride (EtMgCl), propyllithium
(PrLi), propylmagnesium bromide (PrMgBr), propylmagnesium chloride
(PrMgCl), cyclohexanelithium (cyHexLi), cyclohexanemagnesium
bromide (cyHexMgBr), cyclohexanemagnesium chloride (cyHexMgCl), or
combinations thereof.
53. The method of claim 51, wherein the coupling agent is
n-butyllithium (nBuLi), magnesium chloride (MgCl.sub.2),
isopropylmagnesium chloride (iPrMgCl), isopropylmagnesium
chloride-lithium chloride (iPrMgCl--LiCl), tert-butylmagnesium
chloride (tBuMgCl), phenylmagnesium chloride (PhMgCl),
methyllithium (MeLi), methylmagnesium chloride (MeMgCl),
methylmagnesium bromide (MeMgBr), tert-butyllithium (tBuLi),
isopropyllithium (iPrLi), phenyllithium (PhLi), lithium hydride
(LiH), potassium hydride (KH), sodium hydride (NaH), ethyllithium
(EtLi), ethylmagnesium bromide (EtMgBr), ethylmagnesium chloride
(EtMgCl), propyllithium (PrLi), propylmagnesium bromide (PrMgBr),
propylmagnesium chloride (PrMgCl), cyclohexanelithium (cyHexLi),
cyclohexanemagnesium bromide (cyHexMgBr), cyclohexanemagnesium
chloride (cyHexMgCl), or combinations thereof.
54. The method of claim 51, wherein the silylating agent is a
tri-substituted silyl chloride, a tri-substituted silyl bromide, a
tri-substituted silyl iodide, or a tri-substituted silyl
fluoride.
55. The method of claim 51, wherein the deprotonating agent is
PhMgCl; and the coupling agent is iPrMgCl or iPrMgCl--LiCl.
56. The method of claim 51, wherein the deprotonating agent is
PhMgCl; the silylating agent is TMSCl; the coupling agent is
iPrMgCl; and R.sup.b is benzyl.
57. The method of claim 51, wherein the compound of Formula (V-a)
or Formula (V-b) has the structure: ##STR00221## the method
comprising: forming the reaction mixture comprising TMSCl, PhMgCl,
iPrMgCl, the additive, the compound of Formula (VI-a) having
structure: ##STR00222## and the compound of Formula (VII):
##STR00223## to prepare the compound of Formula (V-a) or Formula
(V-b).
58. The method of claim 51, wherein the compound of Formula (V-a)
has the structure: ##STR00224## the method comprising: forming the
reaction mixture comprising TMSCl, PhMgCl, iPrMgCl--LiCl, the
additive, the compound of Formula (VI-a) having structure:
##STR00225## and the compound of Formula (VII): ##STR00226## to
prepare the compound of Formula (V-a).
59. The method of claim 51, wherein the additive is
LaCl.sub.3-2LiCl, LaCl.sub.3, CeCl.sub.3, NdCl.sub.3, or
YCl.sub.3.
60. A compound of formula: ##STR00227## or a pharmaceutically
acceptable salt thereof.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This patent application is a Continuation of U.S. patent
application Ser. No. 14/926,063, filed Oct. 29, 2015, which claims
the benefit under 35 U.S.C. .sctn. 119(e) of U.S. Provisional
Patent Application No. 62/072,331, filed Oct. 29, 2014, and U.S.
Provisional Patent Application No. 62/105,619, filed Jan. 20, 2015.
The foregoing patent applications are incorporated herein by
reference in their entireties.
FIELD OF THE INVENTION
[0002] The invention relates generally to methods and compounds for
treating Filoviridae virus infections, particularly methods and
nucleosides for treating Ebola virus, Marburg virus and Cueva
virus.
BACKGROUND OF THE INVENTION
[0003] Filoviruses (e.g., Ebola virus (EBOV) and Marburg virus
(MARV)) are among the most lethal and destructive viruses. They
cause severe, often fatal viral hemorrhagic fevers in humans and
nonhuman primates (e.g., monkeys, gorillas, and chimpanzees).
Filoviruses are of particular concern as possible biological
weapons since they have the potential for aerosol dissemination and
weaponization.
[0004] The incubation period for Filovirus infection ranges from 2
to 21 days. The onset of illness is abrupt and is characterized by
high fever, headaches, joint and muscle aches, sore throat,
fatigue, diarrhea, vomiting, and stomach pain. A rash, red eyes,
hiccups and internal and external bleeding may be seen in some
patients. Within one week of becoming infected with the virus, most
patients experience chest pains and multiple organ failure, go into
shock, and die. Some patients also experience blindness and
extensive bleeding before dying.
[0005] Filoviridae are a family of RNA viruses. Two members of the
Filoviridae family have been identified: EBOV and MARV. Two key
pathogenic types of the Filoviridae family have been identified:
Ebolavirus and MARV. There is one identified variant of MARV and
five identified species of ebolavirus: Zaire (i.e. Ebola virus,
EBOV), Sudan, Tai Forest, Bundibugyo, and Reston. The exact origin,
locations, and natural habitat of Filoviridae are unknown. However,
on the basis of available evidence and the nature of similar
viruses, it is postulated that Filoviridae are zoonotic (i.e.,
animal-borne) and are normally maintained in an animal host that is
native to the African continent.
[0006] For more than 30 years, ebolaviruses have been associated
with periodic episodes of hemorrhagic fever in Central Africa that
produce severe disease in infected patients. Mortality rates in
outbreaks have ranged from 50% for the Sudan species of ebolavirus
(SEBOV) to up to 90% for the Zaire species of ebolavirus (EBOV,
ZEBOV) (Sanchez et al., Filoviridae: Marburg and Ebola Viruses, in
Fields Virology (eds. Knipe, D. M. & Howley, P. M.) 1409-1448
(Lippincott Williams & Wilkins, Philadelphia)). An outbreak
late in 2007 caused by an apparently new species of ebolavirus in
Uganda resulted in a fatality rate of about 25% (Towner et al.,
PLoS Pathog., 4:e1000212 (2008)). ZEBOV has also decimated
populations of wild apes in this same region of Africa (Walsh et
al., Nature, 422:611-614 (2003)).
[0007] Prevention and treatment of Filovirus infections, including
ebolaviruses (i.e. EBOV) presents many challenges. In fact, there
are no vaccines or post exposure treatment modalities available for
preventing or managing EBOV infections. Patients instead receive
supportive therapy, i.e., electrolyte and fluid balancing, oxygen,
blood pressure maintenance, and treatment for any secondary
infections.
[0008] In view of the importance of novel therapeutics for treating
Filoviridae infections, new efficient methods of producing
ribosides, riboside phosphates and prodrugs are needed.
SUMMARY OF THE INVENTION
[0009] In some embodiments, the present invention provides a method
of preparing a compound of Formula V:
##STR00001##
The method of making the compound of Formula V includes forming a
reaction mixture having a coupling agent, a halo-silane, a compound
of Formula VI:
##STR00002##
and a compound of Formula VII:
##STR00003##
under conditions suitable to prepare the compound of Formula V,
wherein each PG is independently a hydroxy protecting group,
alternatively, two PG groups on adjacent carbons can be combined to
form a --C(R.sup.19).sub.2-- group, R.sup.10 is H or a silyl group,
and R.sup.19 is H, C.sub.1-C.sub.8 alkyl, phenyl or substituted
phenyl.
[0010] In some embodiments, the present invention provides a method
of preparing a compound of Formula V-a or V-b:
##STR00004##
The method of making the compound of Formula V-a or Formula V-b
comprises forming a reaction mixture having a deprotonating agent,
a silylating agent, a coupling agent, an additive, a compound of
Formula VI-a:
##STR00005##
and a compound of Formula VII:
##STR00006##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b, wherein each R.sup.b is independently a hydroxy
protecting group, alternatively, two R.sup.b groups on adjacent
carbons can be combined to form a --C(R.sup.19).sub.2-- group,
R.sup.10 is H or a silyl group, and R.sup.19 is H, C.sub.1-C.sub.8
alkyl, phenyl or substituted phenyl.
[0011] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI:
##STR00007##
wherein R.sup.c is H or a hydroxy protecting group, or two R.sup.c
on adjacent carbons can be combined to form a --C(R.sup.19).sub.2--
group, and R.sup.19 is H or C.sub.1-C.sub.8 alkyl.
[0012] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-a:
##STR00008##
wherein the method comprises forming a reaction mixture having a
cyanating agent, a Lewis Acid, a Broenstedt acid, a solvent, and
the compound of Formula V or V-b:
##STR00009##
under conditions suitable to prepare the compound of Formula XI,
wherein R.sup.b is independently a hydroxy protecting group,
alternatively, two R.sup.b groups on adjacent carbons can be
combined to form a --C(R.sup.19).sub.2 group, R.sup.19 is H or a
silyl group, and R.sup.19 is H, C.sub.1-C.sub.8 alkyl, phenyl or
substituted phenyl.
[0013] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-b:
##STR00010##
wherein the method comprises forming a reaction mixture having a
Lewis Acid, a base, a solvent, a filtering agent, and the compound
of Formula XI-a
##STR00011##
under conditions suitable to prepare the compound of Formula
XI-b.
[0014] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-c:
##STR00012##
wherein the method comprises forming a reaction mixture having a
solvent, a reagent, and the compound of Formula XI-b
##STR00013##
under conditions suitable to prepare the compound of Formula
XI-c.
[0015] In some embodiments, the present invention provides a method
of preparing a compound of Formula VIII:
##STR00014##
wherein the method includes forming a reaction mixture including a
coupling agent, a non-nucleophilic base, a compound of Formula
IX:
##STR00015##
and a compound of Formula X:
##STR00016##
under conditions suitable to form the compound of Formula VIII,
wherein each R.sup.a is H or PG, each PG group is a hydroxy
protecting group, or both PG groups are combined to form
--C(R.sup.19).sub.2--, R.sup.e1 and R.sup.e2 are each independently
H, C.sub.1-C.sub.6 alkyl or benzyl, R.sup.f is H, C.sub.1-C.sub.8
alkyl, benzyl, C.sub.3-C.sub.6 cycloalkyl, or
--CH.sub.2--C.sub.3-C.sub.6 cycloalkyl, R.sup.19 is H,
C.sub.1-C.sub.8 alkyl, phenyl or substituted phenyl, and LG is a
leaving group.
[0016] In some embodiments, the present invention provides a method
of preparing a compound of Formula VIII:
##STR00017##
wherein the method comprises forming a reaction mixture including a
coupling agent, a non-nucleophilic base, a compound of Formula
IX-a:
##STR00018##
and a compound of Formula X:
##STR00019##
under conditions suitable to form the compound of Formula VIII,
wherein R.sup.a is independently H or a hydroxy protecting group,
or two R.sup.a on adjacent carbons can be combined to form a
--C(R.sup.19).sub.2-- group, R.sup.35 is independently H or a
hydroxy protecting group, or two R.sup.35 on adjacent carbons can
be combined to form a --C(R.sup.19).sub.2-- group, R.sup.19 is H or
C.sub.1-C.sub.8 alkyl, R.sup.e1 and R.sup.e2 are each independently
H, C.sub.1-C.sub.6 alkyl or benzyl, R.sup.f is H, C.sub.1-C.sub.8
alkyl, benzyl, C.sub.3-C.sub.6 cycloalkyl, or
--CH.sub.2--C.sub.3-C.sub.6 cycloalkyl, R.sup.19 is H,
C.sub.1-C.sub.8 alkyl, phenyl or substituted phenyl, and LG is a
leaving group.
[0017] In one embodiment, there is provided a method for the
crystallization-induced dynamic resolution of (2S)-2-ethylbutyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate (Formula
X-a):
##STR00020##
to provide (Formula X-b).
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0018] Unless stated otherwise, the following terms and phrases as
used herein are intended to have the following meanings:
[0019] When trade names are used herein, applicants intend to
independently include the trade name product and the active
pharmaceutical ingredient(s) of the trade name product.
[0020] As used herein, "a compound of the invention" or "a compound
of Formula V" means a compound of Formula V or a pharmaceutically
acceptable salt or cocrystal, thereof. In some embodiments, "a
compound of the invention" or "a compound of Formula V" means a
compound of Formula V or a pharmaceutically acceptable salt,
thereof. Similarly, with respect to isolatable intermediates, the
phrase "a compound of Formula (number)" means a compound of that
formula and pharmaceutically acceptable salts or cocrystals,
thereof. In some embodiments, with respect to isolatable
intermediates, the phrase "a compound of Formula (number)" means a
compound of that formula and pharmaceutically acceptable salts,
thereof.
[0021] "Alkyl" is hydrocarbon containing normal, secondary,
tertiary or cyclic carbon atoms. For example, an alkyl group can
have 1 to 20 carbon atoms (i.e, C.sub.1-C.sub.20 alkyl), 1 to 8
carbon atoms (i.e., C.sub.1-C.sub.8 alkyl), or 1 to 6 carbon atoms
(i.e., C.sub.1-C.sub.6 alkyl). Examples of suitable alkyl groups
include, but are not limited to, methyl (Me, --CH.sub.3), ethyl
(Et, --CH.sub.2CH.sub.3), 1-propyl (n-Pr, n-propyl,
--CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl,
--CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu,
--C(CH.sub.3).sub.3), 1-pentyl (n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3, and octyl
(--(CH.sub.2).sub.7CH.sub.3).
[0022] "Alkoxy" means a group having the formula --O-alkyl, in
which an alkyl group, as defined above, is attached to the parent
molecule via an oxygen atom. The alkyl portion of an alkoxy group
can have 1 to 20 carbon atoms (i.e., C.sub.1-C.sub.20 alkoxy), 1 to
12 carbon atoms (i.e., C.sub.1-C.sub.12 alkoxy), or 1 to 6 carbon
atoms (i.e., C.sub.1-C.sub.6 alkoxy). Examples of suitable alkoxy
groups include, but are not limited to, methoxy (--O--CH.sub.3 or
--OMe), ethoxy (--OCH.sub.2CH.sub.3 or --OEt), t-butoxy
(--O--C(CH.sub.3).sub.3 or --OtBu) and the like.
[0023] "Haloalkyl" is an alkyl group, as defined above, in which
one or more hydrogen atoms of the alkyl group is replaced with a
halogen atom. The alkyl portion of a haloalkyl group can have 1 to
20 carbon atoms (i.e., C.sub.1-C.sub.20 haloalkyl), 1 to 12 carbon
atoms (i.e., C.sub.1-C.sub.12 haloalkyl), or 1 to 6 carbon atoms
(i.e., C.sub.1-C.sub.6 alkyl). Examples of suitable haloalkyl
groups include, but are not limited to, --CF.sub.3, --CHF.sub.2,
--CFH.sub.2, --CH.sub.2CF.sub.3, and the like.
[0024] "Alkenyl" is a hydrocarbon containing normal, secondary,
tertiary or cyclic carbon atoms with at least one site of
unsaturation, i.e. a carbon-carbon, sp.sup.2 double bond. For
example, an alkenyl group can have 2 to 20 carbon atoms (i.e.,
C.sub.2-C.sub.20 alkenyl), 2 to 8 carbon atoms (i.e.,
C.sub.2-C.sub.8 alkenyl), or 2 to 6 carbon atoms (i.e.,
C.sub.2-C.sub.6 alkenyl). Examples of suitable alkenyl groups
include, but are not limited to, ethylene or vinyl
(--CH.dbd.CH.sub.2), allyl (--CH.sub.2CH.dbd.CH.sub.2),
cyclopentenyl (--C.sub.5H.sub.7), and 5-hexenyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2).
[0025] "Alkynyl" is a hydrocarbon containing normal, secondary,
tertiary or cyclic carbon atoms with at least one site of
unsaturation, i.e. a carbon-carbon, sp triple bond. For example, an
alkynyl group can have 2 to 20 carbon atoms (i.e., C.sub.2-C.sub.20
alkynyl), 2 to 8 carbon atoms (i.e., C.sub.2-C.sub.8 alkyne,), or 2
to 6 carbon atoms (i.e., C.sub.2-C.sub.6 alkynyl). Examples of
suitable alkynyl groups include, but are not limited to, acetylenic
(--C.ident.CH), propargyl (--CH.sub.2C.ident.CH), and the like.
[0026] "Alkylene" refers to a saturated, branched or straight chain
or cyclic hydrocarbon radical having two monovalent radical centers
derived by the removal of two hydrogen atoms from the same or two
different carbon atoms of a parent alkane. For example, an alkylene
group can have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to
6 carbon atoms. Typical alkylene radicals include, but are not
limited to, methylene (--CH.sub.2--), 1,1-ethyl (--CH(CH.sub.3)--),
1,2-ethyl (--CH.sub.2CH.sub.2--), 1,1-propyl
(--CH(CH.sub.2CH.sub.3)--), 1,2-propyl (--CH.sub.2CH(CH.sub.3)--),
1,3-propyl (--CH.sub.2CH.sub.2CH.sub.2--), 1,4-butyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), and the like.
[0027] "Alkenylene" refers to an unsaturated, branched or straight
chain or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkene. For example, and
alkenylene group can have 1 to 20 carbon atoms, 1 to 10 carbon
atoms, or 1 to 6 carbon atoms. Typical alkenylene radicals include,
but are not limited to, 1,2-ethylene (--CH.dbd.CH--).
[0028] "Alkynylene" refers to an unsaturated, branched or straight
chain or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkyne. For example, an
alkynylene group can have 1 to 20 carbon atoms, 1 to 10 carbon
atoms, or 1 to 6 carbon atoms. Typical alkynylene radicals include,
but are not limited to, acetylene (--C.ident.C--), propargyl
(--CH.sub.2C.ident.C--), and 4-pentynyl
(--CH.sub.2CH.sub.2CH.sub.2C.ident.C--).
[0029] "Amino" refers generally to a nitrogen radical which can be
considered a derivative of ammonia, having the formula
--N(X).sub.2, where each "X" is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted carbocyclyl,
substituted or unsubstituted heterocyclyl, etc. The hybridization
of the nitrogen is approximately sp.sup.3. Nonlimiting types of
amino include --NH.sub.2, --N(alkyl).sub.2, --NH(alkyl),
--N(carbocyclyl).sub.2, --NH(carbocyclyl), --N(heterocyclyl).sub.2,
--NH(heterocyclyl), --N(aryl).sub.2, --NH(aryl), --N(alkyl)(aryl),
--N(alkyl)(heterocyclyl), --N(carbocyclyl)(heterocyclyl),
--N(aryl)(heteroaryl), --N(alkyl)(heteroaryl), etc. The term
"alkylamino" refers to an amino group substituted with at least one
alkyl group. Nonlimiting examples of amino groups include
--NH.sub.2, --NH(CH.sub.3), --N(CH.sub.3).sub.2,
--NH(CH.sub.2CH.sub.3), --N(CH.sub.2CH.sub.3).sub.2, --NH(phenyl),
--N(phenyl).sub.2, --NH(benzyl), --N(benzyl).sub.2, etc.
Substituted alkylamino refers generally to alkylamino groups, as
defined above, in which at least one substituted alkyl, as defined
herein, is attached to the amino nitrogen atom. Non-limiting
examples of substituted alkylamino includes
--NH(alkylene-C(O)--OH), --NH(alkylene-C(O)--O-alkyl),
--N(alkylene-C(O)--OH).sub.2, --N(alkylene-C(O)--O-alkyl).sub.2,
etc.
[0030] "Aryl" means an aromatic hydrocarbon radical derived by the
removal of one hydrogen atom from a single carbon atom of a parent
aromatic ring system. For example, an aryl group can have 6 to 20
carbon atoms, 6 to 14 carbon atoms, or 6 to 10 carbon atoms.
Typical aryl groups include, but are not limited to, radicals
derived from benzene (e.g., phenyl), substituted benzene,
naphthalene, anthracene, biphenyl, and the like. Further typical
aryl groups include, but are not limited to, phenyl.
[0031] "Arylalkyl" refers to an acyclic alkyl radical in which one
of the hydrogen atoms bonded to a carbon atom, typically a terminal
or sp.sup.3 carbon atom, is replaced with an aryl radical. Typical
arylalkyl groups include, but are not limited to, benzyl,
2-phenylethan-1-yl, naphthylmethyl, 2-naphthylethan-1-yl,
naphthobenzyl, 2-naphthophenylethan-1-yl and the like. The
arylalkyl group can comprise 7 to 20 carbon atoms, e.g., the alkyl
moiety is 1 to 6 carbon atoms and the aryl moiety is 6 to 14 carbon
atoms.
[0032] "Arylalkenyl" refers to an acyclic alkenyl radical in which
one of the hydrogen atoms bonded to a carbon atom, typically a
terminal or sp.sup.3 carbon atom, but also an sp.sup.2 carbon atom,
is replaced with an aryl radical. The aryl portion of the
arylalkenyl can include, for example, any of the aryl groups
disclosed herein, and the alkenyl portion of the arylalkenyl can
include, for example, any of the alkenyl groups disclosed herein.
The arylalkenyl group can comprise 8 to 20 carbon atoms, e.g., the
alkenyl moiety is 2 to 6 carbon atoms and the aryl moiety is 6 to
14 carbon atoms.
[0033] "Arylalkynyl" refers to an acyclic alkynyl radical in which
one of the hydrogen atoms bonded to a carbon atom, typically a
terminal or sp.sup.3 carbon atom, but also an sp carbon atom, is
replaced with an aryl radical. The aryl portion of the arylalkynyl
can include, for example, any of the aryl groups disclosed herein,
and the alkynyl portion of the arylalkynyl can include, for
example, any of the alkynyl groups disclosed herein. The
arylalkynyl group can comprise 8 to 20 carbon atoms, e.g., the
alkynyl moiety is 2 to 6 carbon atoms and the aryl moiety is 6 to
14 carbon atoms.
[0034] The term "substituted" in reference to alkyl, alkylene,
aryl, arylalkyl, alkoxy, heterocyclyl, heteroaryl, carbocyclyl,
etc., for example, "substituted alkyl", "substituted alkylene",
"substituted aryl", "substituted arylalkyl", "substituted
heterocyclyl", and "substituted carbocyclyl" means alkyl, alkylene,
aryl, arylalkyl, heterocyclyl, carbocyclyl respectively, in which
one or more hydrogen atoms are each independently replaced with a
non-hydrogen substituent. The term "substituted phenyl" means
phenyl, in which one or more hydrogen atoms are each independently
replaced with a non-hydrogen substituent. Typical substituents
include, but are not limited to, --X, --R.sup.b, --O.sup.-, .dbd.O,
--OR.sup.b, --SR.sup.b, --S.sup.-, --NR.sup.b.sub.2,
--N.sup.+R.sup.b.sub.3, =NR.sup.b, --CX.sub.3, --CN, --OCN, --SCN,
--N.dbd.C.dbd.O, --NCS, --NO, --NO.sub.2, .dbd.N.sub.2, --N.sub.3,
--NHC(.dbd.O)R.sup.b, --OC(.dbd.O)R.sup.b,
--NHC(.dbd.O)NR.sup.b.sub.2, --S(.dbd.O).sub.2--,
--S(.dbd.O).sub.2OH, --S(.dbd.O).sub.2R.sup.b,
--OS(.dbd.O).sub.2OR.sup.b, --S(.dbd.O).sub.2NR.sup.b.sub.2,
--S(.dbd.O)R.sup.b, --OP(.dbd.O)(OR.sup.b).sub.2,
--P(.dbd.O)(OR.sup.b).sub.2, --P(.dbd.O)(O.sup.-).sub.2,
--P(.dbd.O)(OH).sub.2, --P(O)(OR.sup.b)(O), --C(.dbd.O)R.sup.b,
--C(.dbd.O)X, --C(S)R.sup.b, --C(O)OR.sup.b, --C(O)O.sup.-,
--C(S)OR.sup.b, --C(O)SR.sup.b, --C(S)SR.sup.b,
--C(O)NR.sup.b.sub.2, --C(S)NR.sup.b.sub.2,
--C(.dbd.NR.sup.b)NR.sup.b.sub.2, where each X is independently a
halogen: F, Cl, Br, or I; and each R.sup.b is independently H,
alkyl, aryl, arylalkyl, a heterocycle, or a protecting group or
prodrug moiety. Alkylene, alkenylene, and alkynylene groups may
also be similarly substituted. Unless otherwise indicated, when the
term "substituted" is used in conjunction with groups such as
arylalkyl, which have two or more moieties capable of substitution,
the substituents can be attached to the aryl moiety, the alkyl
moiety, or both.
[0035] The term "prodrug" as used herein refers to any compound
that when administered to a biological system generates the drug
substance, i.e., active ingredient, as a result of spontaneous
chemical reaction(s), enzyme catalyzed chemical reaction(s),
photolysis, and/or metabolic chemical reaction(s). A prodrug is
thus a covalently modified analog or latent form of a
therapeutically active compound.
[0036] One skilled in the art will recognize that substituents and
other moieties of the compounds of Formula I-IV should be selected
in order to provide a compound which is sufficiently stable to
provide a pharmaceutically useful compound which can be formulated
into an acceptably stable pharmaceutical composition. Compounds of
Formula I-IV which have such stability are contemplated as falling
within the scope of the present invention.
[0037] "Heteroalkyl" refers to an alkyl group where one or more
carbon atoms have been replaced with a heteroatom, such as, O, N,
or S. For example, if the carbon atom of the alkyl group which is
attached to the parent molecule is replaced with a heteroatom
(e.g., O, N, or S) the resulting heteroalkyl groups are,
respectively, an alkoxy group (e.g., --OCH.sub.3, etc.), an amine
(e.g., --NHCH.sub.3, --N(CH.sub.3).sub.2, etc.), or a thioalkyl
group (e.g., --SCH.sub.3). If a non-terminal carbon atom of the
alkyl group which is not attached to the parent molecule is
replaced with a heteroatom (e.g., O, N, or S) the resulting
heteroalkyl groups are, respectively, an alkyl ether (e.g.,
--CH.sub.2CH.sub.2--O--CH.sub.3, etc.), an alkyl amine (e.g.,
--CH.sub.2NHCH.sub.3, --CH.sub.2N(CH.sub.3).sub.2, etc.), or a
thioalkyl ether (e.g., --CH.sub.2--S--CH.sub.3). If a terminal
carbon atom of the alkyl group is replaced with a heteroatom (e.g.,
O, N, or S), the resulting heteroalkyl groups are, respectively, a
hydroxyalkyl group (e.g., --CH.sub.2CH.sub.2--OH), an aminoalkyl
group (e.g., --CH.sub.2NH.sub.2), or an alkyl thiol group (e.g.,
--CH.sub.2CH.sub.2--SH). A heteroalkyl group can have, for example,
1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms.
A C.sub.1-C.sub.6 heteroalkyl group means a heteroalkyl group
having 1 to 6 carbon atoms.
[0038] "Heterocycle" or "heterocyclyl" as used herein includes by
way of example and not limitation those heterocycles described in
Paquette, Leo A.; Principles of Modern Heterocyclic Chemistry (W.
A. Benjamin, New York, 1968), particularly Chapters 1, 3, 4, 6, 7,
and 9; The Chemistry of Heterocyclic Compounds, A Series of
Monographs" (John Wiley & Sons, New York, 1950 to present), in
particular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc.
(1960) 82:5566. In one specific embodiment of the invention
"heterocycle" includes a "carbocycle" as defined herein, wherein
one or more (e.g. 1, 2, 3, or 4) carbon atoms have been replaced
with a heteroatom (e.g. O, N, or S). The terms "heterocycle" or
"heterocyclyl" includes saturated rings, partially unsaturated
rings, and aromatic rings (i.e., heteroaromatic rings). Substituted
heterocyclyls include, for example, heterocyclic rings substituted
with any of the substituents disclosed herein including carbonyl
groups. A non-limiting example of a carbonyl substituted
heterocyclyl is:
##STR00021##
[0039] Examples of heterocycles include by way of example and not
limitation pyridyl, dihydroypyridyl, tetrahydropyridyl (piperidyl),
thiazolyl, tetrahydrothiophenyl, sulfur oxidized
tetrahydrothiophenyl, pyrimidinyl, furanyl, thienyl, pyrrolyl,
pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl,
indolyl, indolenyl, quinolinyl, isoquinolinyl, benzimidazolyl,
piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl,
pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, decahydroquinolinyl,
octahydroisoquinolinyl, azocinyl, triazinyl, 6H-1,2,5-thiadiazinyl,
2H,6H-1,5,2-dithiazinyl, thienyl, thianthrenyl, pyranyl,
isobenzofuranyl, chromenyl, xanthenyl, phenoxathinyl, 2H-pyrrolyl,
isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl,
isoindolyl, 3H-indolyl, 1H-indazoly, purinyl, 4H-quinolizinyl,
phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl,
cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl,
.beta.-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl,
phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl,
phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl,
imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl,
isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl,
benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl,
isatinoyl, and bis-tetrahydrofuranyl:
##STR00022##
[0040] By way of example and not limitation, carbon bonded
heterocycles are bonded at position 2, 3, 4, 5, or 6 of a pyridine,
position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a
pyrimidine, position 2, 3, 5, or 6 of a pyrazine, position 2, 3, 4,
or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or
tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or
thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or
isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4
of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or
position 1, 3, 4, 5, 6, 7, or 8 of an isoquinoline. Still more
typically, carbon bonded heterocycles include 2-pyridyl, 3-pyridyl,
4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl, 4-pyridazinyl,
5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl, 4-pyrimidinyl,
5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl,
5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or
5-thiazolyl.
[0041] By way of example and not limitation, nitrogen bonded
heterocycles are bonded at position 1 of an aziridine, azetidine,
pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole,
imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline,
2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole,
indoline, 1H-indazole, position 2 of a isoindole, or isoindoline,
position 4 of a morpholine, and position 9 of a carbazole, or
.beta.-carboline. Still more typically, nitrogen bonded
heterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl,
1-imidazolyl, 1-pyrazolyl, and 1-piperidinyl.
[0042] "Heterocyclylalkyl" refers to an acyclic alkyl radical in
which one of the hydrogen atoms bonded to a carbon atom, typically
a terminal or sp.sup.3 carbon atom, is replaced with a heterocyclyl
radical (i.e., a heterocyclyl-alkylene- moiety). Typical
heterocyclyl alkyl groups include, but are not limited to
heterocyclyl-CH.sub.2--, 2-(heterocyclyl)ethan-1-yl, and the like,
wherein the "heterocyclyl" portion includes any of the heterocyclyl
groups described above, including those described in Principles of
Modern Heterocyclic Chemistry. One skilled in the art will also
understand that the heterocyclyl group can be attached to the alkyl
portion of the heterocyclyl alkyl by means of a carbon-carbon bond
or a carbon-heteroatom bond, with the proviso that the resulting
group is chemically stable. The heterocyclyl alkyl group comprises
3 to 20 carbon atoms, e.g., the alkyl portion of the arylalkyl
group is 1 to 6 carbon atoms and the heterocyclyl moiety is 2 to 14
carbon atoms. Examples of heterocyclylalkyls include by way of
example and not limitation 5-membered sulfur, oxygen, and/or
nitrogen containing heterocycles such as thiazolylmethyl,
2-thiazolylethan-1-yl, imidazolylmethyl, oxazolylmethyl,
thiadiazolylmethyl, etc., 6-membered sulfur, oxygen, and/or
nitrogen containing heterocycles such as piperidinylmethyl,
piperazinylmethyl, morpholinylmethyl, pyridinylmethyl,
pyridizylmethyl, pyrimidylmethyl, pyrazinylmethyl, etc.
[0043] "Heterocyclylalkenyl" refers to an acyclic alkenyl radical
in which one of the hydrogen atoms bonded to a carbon atom,
typically a terminal or sp.sup.3 carbon atom, but also a sp.sup.2
carbon atom, is replaced with a heterocyclyl radical (i.e., a
heterocyclyl-alkenylene- moiety). The heterocyclyl portion of the
heterocyclyl alkenyl group includes any of the heterocyclyl groups
described herein, including those described in Principles of Modern
Heterocyclic Chemistry, and the alkenyl portion of the heterocyclyl
alkenyl group includes any of the alkenyl groups disclosed herein.
One skilled in the art will also understand that the heterocyclyl
group can be attached to the alkenyl portion of the heterocyclyl
alkenyl by means of a carbon-carbon bond or a carbon-heteroatom
bond, with the proviso that the resulting group is chemically
stable. The heterocyclyl alkenyl group comprises 4 to 20 carbon
atoms, e.g., the alkenyl portion of the heterocyclyl alkenyl group
is 2 to 6 carbon atoms and the heterocyclyl moiety is 2 to 14
carbon atoms.
[0044] "Heterocyclylalkynyl" refers to an acyclic alkynyl radical
in which one of the hydrogen atoms bonded to a carbon atom,
typically a terminal or sp.sup.3 carbon atom, but also an sp carbon
atom, is replaced with a heterocyclyl radical (i.e., a
heterocyclyl-alkynylene- moiety). The heterocyclyl portion of the
heterocyclyl alkynyl group includes any of the heterocyclyl groups
described herein, including those described in Principles of Modern
Heterocyclic Chemistry, and the alkynyl portion of the heterocyclyl
alkynyl group includes any of the alkynyl groups disclosed herein.
One skilled in the art will also understand that the heterocyclyl
group can be attached to the alkynyl portion of the heterocyclyl
alkynyl by means of a carbon-carbon bond or a carbon-heteroatom
bond, with the proviso that the resulting group is chemically
stable. The heterocyclyl alkynyl group comprises 4 to 20 carbon
atoms, e.g., the alkynyl portion of the heterocyclyl alkynyl group
is 2 to 6 carbon atoms and the heterocyclyl moiety is 2 to 14
carbon atoms.
[0045] "Heteroaryl" refers to an aromatic heterocyclyl having at
least one heteroatom in the ring. Non-limiting examples of suitable
heteroatoms which can be included in the aromatic ring include
oxygen, sulfur, and nitrogen. Non-limiting examples of heteroaryl
rings include all of those aromatic rings listed in the definition
of "heterocyclyl", including pyridinyl, pyrrolyl, oxazolyl,
indolyl, isoindolyl, purinyl, furanyl, thienyl, benzofuranyl,
benzothiophenyl, carbazolyl, imidazolyl, thiazolyl, isoxazolyl,
pyrazolyl, isothiazolyl, quinolyl, isoquinolyl, pyridazyl,
pyrimidyl, pyrazyl, etc.
[0046] "Carbocycle" or "carbocyclyl" refers to a saturated (i.e.,
cycloalkyl), partially unsaturated (e.g., cycloakenyl,
cycloalkadienyl, etc.) or aromatic ring having 3 to 7 carbon atoms
as a monocycle, 7 to 12 carbon atoms as a bicycle, and up to about
20 carbon atoms as a polycycle. Monocyclic carbocycles have 3 to 7
ring atoms, still more typically 5 or 6 ring atoms. Bicyclic
carbocycles have 7 to 12 ring atoms, e.g., arranged as a bicyclo
[4,5], [5,5], [5,6] or [6,6] system, or 9 or 10 ring atoms arranged
as a bicyclo [5,6] or [6,6] system, or spiro-fused rings.
Non-limiting examples of monocyclic carbocycles include
cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl,
1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,
1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, and
phenyl. Non-limiting examples of bicyclo carbocycles includes
naphthyl, tetrahydronapthalene, and decaline.
[0047] "Carbocyclylalkyl" refers to an acyclic akyl radical in
which one of the hydrogen atoms bonded to a carbon atom is replaced
with a carbocyclyl radical as described herein. Typical, but
non-limiting, examples of carbocyclylalkyl groups include
cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl,
cyclopentylmethyl and cyclohexylmethyl.
[0048] "Arylheteroalkyl" refers to a heteroalkyl as defined herein,
in which a hydrogen atom (which may be attached either to a carbon
atom or a heteroatom) has been replaced with an aryl group as
defined herein. The aryl groups may be bonded to a carbon atom of
the heteroalkyl group, or to a heteroatom of the heteroalkyl group,
provided that the resulting arylheteroalkyl group provides a
chemically stable moiety. For example, an arylheteroalkyl group can
have the general formulae -alkylene-O-aryl,
-alkylene-O-alkylene-aryl, -alkylene-NH-aryl,
-alkylene-NH-alkylene-aryl, -alkylene-S-aryl,
-alkylene-S-alkylene-aryl, etc. In addition, any of the alkylene
moieties in the general formulae above can be further substituted
with any of the substituents defined or exemplified herein.
[0049] "Heteroarylalkyl" refers to an alkyl group, as defined
herein, in which a hydrogen atom has been replaced with a
heteroaryl group as defined herein. Non-limiting examples of
heteroaryl alkyl include --CH.sub.2-pyridinyl, --CH.sub.2-pyrrolyl,
--CH.sub.2-oxazolyl, --CH.sub.2-indolyl, --CH.sub.2-isoindolyl,
--CH.sub.2-purinyl, --CH.sub.2-furanyl, --CH.sub.2-thienyl,
--CH.sub.2-benzofuranyl, --CH.sub.2-benzothiophenyl,
--CH.sub.2-carbazolyl, --CH.sub.2-imidazolyl, --CH.sub.2-thiazolyl,
--CH.sub.2-isoxazolyl, --CH.sub.2-pyrazolyl,
--CH.sub.2-isothiazolyl, --CH.sub.2-quinolyl,
--CH.sub.2-isoquinolyl, --CH.sub.2-pyridazyl, --CH.sub.2-pyrimidyl,
--CH.sub.2-pyrazyl, --CH(CH.sub.3)-pyridinyl,
--CH(CH.sub.3)-pyrrolyl, --CH(CH.sub.3)-oxazolyl,
--CH(CH.sub.3)-indolyl, --CH(CH.sub.3)-isoindolyl,
--CH(CH.sub.3)-purinyl, --CH(CH.sub.3)-furanyl,
--CH(CH.sub.3)-thienyl, --CH(CH.sub.3)-benzofuranyl,
--CH(CH.sub.3)-benzothiophenyl, --CH(CH.sub.3)-carbazolyl,
--CH(CH.sub.3)-imidazolyl, --CH(CH.sub.3)-thiazolyl,
--CH(CH.sub.3)-isoxazolyl, --CH(CH.sub.3)-pyrazolyl,
--CH(CH.sub.3)-isothiazolyl, --CH(CH.sub.3)-quinolyl,
--CH(CH.sub.3)-isoquinolyl, --CH(CH.sub.3)-pyridazyl,
--CH(CH.sub.3)-pyrimidyl, --CH(CH.sub.3)-pyrazyl, etc.
[0050] The term "optionally substituted" in reference to a
particular moiety of the compound of Formula I-IV (e.g., an
optionally substituted aryl group) refers to a moiety wherein all
substituents are hydrogen or wherein one or more of the hydrogens
of the moiety may be replaced by substituents such as those listed
under the definition of "substituted".
[0051] The term "optionally replaced" in reference to a particular
moiety of the compound of Formula I-IV (e.g., the carbon atoms of
said (C.sub.1-C.sub.8)alkyl may be optionally replaced by --O--,
--S--, or --NR.sup.a--) means that one or more of the methylene
groups of the (C.sub.1-C.sub.8)alkyl may be replaced by 0, 1, 2, or
more of the groups specified (e.g., --O--, --S--, or
--NR.sup.a--).
[0052] The term "non-terminal carbon atom(s)" in reference to an
alkyl, alkenyl, alkynyl, alkylene, alkenylene, or alkynylene moiety
refers to the carbon atoms in the moiety that intervene between the
first carbon atom of the moiety and the last carbon atom in the
moiety. Therefore, by way of example and not limitation, in the
alkyl moiety --CH.sub.2(C*)H.sub.2(C*)H.sub.2CH.sub.3 or alkylene
moiety --CH.sub.2(C*)H.sub.2(C*)H.sub.2CH.sub.2-- the C* atoms
would be considered to be the non-terminal carbon atoms.
[0053] Certain Q and Q.sup.1 alternatives are nitrogen oxides such
as .sup.+N(O)(R) or .sup.+N(O)(OR). These nitrogen oxides, as shown
here attached to a carbon atom, can also be represented by charge
separated groups such as
##STR00023##
respectively, and are intended to be equivalent to the
aforementioned representations for the purposes of describing this
invention.
[0054] "Linker" or "link" means a chemical moiety comprising a
covalent bond or a chain of atoms. Linkers include repeating units
of alkyloxy (e.g. polyethyleneoxy, PEG, polymethyleneoxy) and
alkylamino (e.g. polyethyleneamino, Jeffamine.TM.); and diacid
ester and amides including succinate, succinamide, diglycolate,
malonate, and caproamide.
[0055] The terms such as "oxygen-linked", "nitrogen-linked",
"carbon-linked", "sulfur-linked", or "phosphorous-linked" mean that
if a bond between two moieties can be formed by using more than one
type of atom in a moiety, then the bond formed between the moieties
is through the atom specified. For example, a nitrogen-linked amino
acid would be bonded through a nitrogen atom of the amino acid
rather than through an oxygen or carbon atom of the amino acid.
[0056] In some embodiments of the compounds of Formula I-IV, one or
more of Z.sup.1 or Z.sup.2 are independently a radical of a
nitrogen-linked naturally occurring .alpha.-amino acid ester.
Examples of naturally occurring amino acids include isoleucine,
leucine, lysine, methionine, phenylalanine, threonine, tryptophan,
valine, alanine, asparagine, aspartic acid, cysteine, glutamic
acid, glutamine, glycine, proline, selenocysteine, serine,
tyrosine, arginine, histidine, ornithine and taurine. The esters of
these amino acids comprise any of those described for the
substituent R, particularly those in which R is optionally
substituted (C.sub.1-C.sub.8)alkyl.
[0057] The term "purine" or "pyrimidine" base comprises, but is not
limited to, adenine, N.sup.6-alkylpurines, N.sup.6-acylpurines
(wherein acyl is C(O)(alkyl, aryl, alkylaryl, or arylalkyl),
N.sup.6-benzylpurine, N.sup.6-halopurine, N.sup.6-vinylpurine,
N.sup.6-acetylenic purine, N.sup.6-acyl purine,
N.sup.6-hydroxyalkyl purine, N.sup.6-allylaminopurine,
N.sup.6-thioallyl purine, N.sup.2-alkylpurines,
N.sup.2-alkyl-6-thiopurines, thymine, cytosine, 5-fluorocytosine,
5-methylcytosine, 6-azapyrimidine, including 6-azacytosine, 2-
and/or 4-mercaptopyrmidine, uracil, 5-halouracil, including
5-fluorouracil, C.sup.5-alkylpyrimidines,
C.sup.5-benzylpyrimidines, C.sup.5-halopyrimidines,
C.sup.5-vinylpyrimidine, C.sup.5-acetylenic pyrimidine,
C.sup.5-acyl pyrimidine, C.sup.5-hydroxyalkyl purine,
C.sup.5-amidopyrimidine, C.sup.5-cyanopyrimidine,
C.sup.5-5-iodopyrimidine, C.sup.6-iodo-pyrimidine,
C.sup.5--Br-vinyl pyrimidine, C.sup.6--Br-- vinyl pyriniidine,
C.sup.5-nitropyrimidine, C.sup.5-amino-pyrimidine,
N.sup.2-alkylpurines, N.sup.2-alkyl-6-thiopurines, 5-azacytidinyl,
5-azauracilyl, triazolopyridinyl, imidazolopyridinyl,
pyrrolopyrimidinyl, and pyrazolopyrimidinyl. Purine bases include,
but are not limited to, guanine, adenine, hypoxanthine,
2,6-diaminopurine, and 6-chloropurine. The purine and pyrimidine
bases are linked to the ribose sugar, or analog thereof, through a
nitrogen atom of the base. Functional oxygen and nitrogen groups on
the base can be protected as necessary or desired. Suitable
protecting groups are well known to those skilled in the art, and
include trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl,
and t-butyldiphenylsilyl, trityl, alkyl groups, and acyl groups
such as acetyl and propionyl, methanesulfonyl, and
p-toluenesulfonyl.
[0058] Unless otherwise specified, the carbon atoms of the
compounds of Formula I-IV are intended to have a valence of four.
In some chemical structure representations where carbon atoms do
not have a sufficient number of variables attached to produce a
valence of four, the remaining carbon substituents needed to
provide a valence of four should be assumed to be hydrogen. For
example,
##STR00024##
has the same meaning as
##STR00025##
[0059] "Protecting group" refers to a moiety of a compound that
masks or alters the properties of a functional group or the
properties of the compound as a whole. The chemical substructure of
a protecting group varies widely. One function of a protecting
group is to serve as an intermediate in the synthesis of the
parental drug substance. Chemical protecting groups and strategies
for protection/deprotection are well known in the art. See:
"Protective Groups in Organic Chemistry", Theodora W. Greene (John
Wiley & Sons, Inc., New York, 1991. See also Protective Groups
in Organic Chemistry, Peter G. M. Wuts and Theodora W. Greene, 4th
Ed., 2006. Protecting groups are often utilized to mask the
reactivity of certain functional groups, to assist in the
efficiency of desired chemical reactions, e.g. making and breaking
chemical bonds in an ordered and planned fashion. Protection of
functional groups of a compound alters other physical properties
besides the reactivity of the protected functional group, such as
the polarity, lipophilicity (hydrophobicity), and other properties
which can be measured by common analytical tools. Chemically
protected intermediates may themselves be biologically active or
inactive. "Hydroxy protecting groups" refers to those protecting
groups useful for protecting hydroxy groups (--OH).
[0060] Protected compounds may also exhibit altered, and in some
cases, optimized properties in vitro and in vivo, such as passage
through cellular membranes and resistance to enzymatic degradation
or sequestration. In this role, protected compounds with intended
therapeutic effects may be referred to as prodrugs. Another
function of a protecting group is to convert the parental drug into
a prodrug, whereby the parental drug is released upon conversion of
the prodrug in vivo. Because active prodrugs may be absorbed more
effectively than the parental drug, prodrugs may possess greater
potency in vivo than the parental drug. Protecting groups are
removed either in vitro, in the instance of chemical intermediates,
or in vivo, in the case of prodrugs. With chemical intermediates,
it is not particularly important that the resulting products after
deprotection, e.g. alcohols, be physiologically acceptable,
although in general it is more desirable if the products are
pharmacologically innocuous.
[0061] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0062] The term "stereoisomers" refers to compounds which have
identical chemical constitution, but differ with regard to the
arrangement of the atoms or groups in space.
[0063] "Diastereomer" refers to a stereoisomer with two or more
centers of chirality and whose molecules are not mirror images of
one another. Diastereomers have different physical properties, e.g.
melting points, boiling points, spectral properties, reactivities
and biological properties. For example, the compounds of Formula
I-IV may have a chiral phosphorus atom when R.sup.7 is
##STR00026##
and Z.sup.1 and Z.sup.2 are different. When at least one of either
Z.sup.1 or Z.sup.2 also has a chiral center, for example with
Z.sup.1 or Z.sup.2 is a nitrogen-linked, chiral, naturally
occurring .alpha.-amino acid ester, then the compound of Formula
I-IV will exists as diastereomers because there are two centers of
chirality in the molecule. All such diastereomers and their uses
described herein are encompassed by the instant invention. Mixtures
of diastereomers may be separate under high resolution analytical
procedures such as electrophoresis, crystallization and/or
chromatography. Diastereomers may have different physical
attributes such as, but not limited to, solubility, chemical
stabilities and crystallinity and may also have different
biological properties such as, but not limited to, enzymatic
stability, absorption and metabolic stability.
[0064] "Enantiomers" refer to two stereoisomers of a compound which
are non-superimposable mirror images of one another.
[0065] The modifier "about" used in connection with a quantity is
inclusive of the stated value and has the meaning dictated by the
context (e.g., includes the degree of error associated with
measurement of the particular quantity).
[0066] The term "treating", as used herein, unless otherwise
indicated, means reversing, alleviating, inhibiting the progress
of, or preventing the disorder or condition to which such term
applies, or one or more symptoms of such disorder or condition. The
term "treatment", as used herein, refers to the act of treating, as
"treating" is defined immediately above.
[0067] The term "therapeutically effective amount", as used herein,
is the amount of compound of Formula I-IV present in a composition
described herein that is needed to provide a desired level of drug
in the secretions and tissues of the airways and lungs, or
alternatively, in the bloodstream of a subject to be treated to
give an anticipated physiological response or desired biological
effect when such a composition is administered by the chosen route
of administration. The precise amount will depend upon numerous
factors, for example the particular compound of Formula I-IV, the
specific activity of the composition, the delivery device employed,
the physical characteristics of the composition, its intended use,
as well as patient considerations such as severity of the disease
state, patient cooperation, etc., and can readily be determined by
one skilled in the art based upon the information provided
herein.
[0068] The term "normal saline" means a water solution containing
0.9% (w/v) NaCl.
[0069] The term "hypertonic saline" means a water solution
containing greater than 0.9% (w/v) NaCl. For example, 3% hypertonic
saline would contain 3% (w/v) NaCl.
[0070] "Forming a reaction mixture" refers to the process of
bringing into contact at least two distinct species such that they
mix together and can react. It should be appreciated, however, the
resulting reaction product can be produced directly from a reaction
between the added reagents or from an intermediate from one or more
of the added reagents which can be produced in the reaction
mixture.
[0071] "Coupling agent" refers to an agent capable of coupling two
disparate compounds. Coupling agents can be catalytic or
stoichiometric. For example, the coupling agents can be a lithium
based coupling agent or a magnesium based coupling agent such as a
Grignard reagent. Exemplary coupling agents include, but are not
limited to, n-BuLi, MgCl.sub.2, iPrMgCl, tBuMgCl, PhMgCl or
combinations thereof.
[0072] "Silane" refers to a silicon containing group having the
formula SiR.sub.4, where each R group can be alkyl, alkenyl,
cycloalkyl, phenyl, or other silicon containing groups. When the
silane is linked to another compound, the silane is referred to as
a "silyl" and has the formula --SiR.sub.3.
[0073] "Halo-silane" refers to a silane having at least one halogen
group linked to the silicon atom. Representative halo-silanes have
the formula Halo-SiR.sub.3, where each R group can be alkyl,
alkenyl, cycloalkyl, phenyl, or other silicon containing groups.
Specific halo-silanes include Cl--Si(CH.sub.3).sub.3, and
Cl--Si(CH.sub.3).sub.2CH.sub.2CH.sub.2Si(CH.sub.3).sub.2--Cl.
[0074] "Non-nucleophilic base" refers to an electron donor, a Lewis
base, such as nitrogen bases including triethylamine,
diisopropylethyl amine, N,N-diethylaniline, pyridine, 2,6-lutidine,
2,4,6-collidine, 4-dimethylaminopyridine, and quinuclidine.
[0075] "Leaving group" refers to groups that maintain the bonding
electron pair during heterolytic bond cleavage. For example, a
leaving group is readily displaced during a nucleophilic
displacement reaction. Suitable leaving groups include, but are not
limited to, chloride, bromide, mesylate, tosylate, triflate,
4-nitrobenzenesulfonate, 4-chlorobenzenesulfonate, 4-nitrophenoxy,
pentafluorophenoxy, etc. One of skill in the art will recognize
other leaving groups useful in the present invention.
[0076] "Deprotection agent" refers to any agent capable of removing
a protecting group. The deprotection agent will depend on the type
of protecting group used. Representative deprotection agents are
known in the art and can be found in Protective Groups in Organic
Chemistry, Peter G. M. Wuts and Theodora W. Greene, 4th Ed.,
2006.
II. Preparation of Compounds
[0077] The compounds of the present invention can be prepared by a
variety of means. For example, protected nucleosides of Formula V
can be prepared by reaction of a protected lactone with an
iodo-substituted base under suitable coupling conditions. The
nucleosides can then be modified with a prodrug moiety by reaction
of a partially protected nucleoside with a suitable prodrug moiety,
following be removal of the protecting groups, to afford the
compounds of the present invention.
[0078] A. Preparation of Nucleosides Via Iodo-Base
[0079] In one embodiment, the present invention provides a method
of preparing a compound of Formula V:
##STR00027##
The method of making the compound of Formula V includes forming a
reaction mixture having a coupling agent, a halo-silane, a compound
of Formula VI:
##STR00028##
and a compound of Formula VII:
##STR00029##
under conditions suitable to prepare the compound of Formula V,
wherein each PG is independently a hydroxy protecting group,
alternatively, two PG groups on adjacent carbons can be combined to
form a --C(R.sup.19).sub.2-- group, R.sup.10 is H or a silyl group,
and R.sup.19 is H, C.sub.1-C.sub.8 alkyl, phenyl or substituted
phenyl.
[0080] Any suitable coupling agent can be used in the method of
making the compound of Formula V. The coupling agent can be a
lithium coupling agent, a sodium coupling agent, a magnesium
coupling agent, or others. For example, the coupling agent can be a
deprotonating agent such as n-butyl lithium (n-BuLi), sodium
hydride (NaH), lithium aluminum hydride (LAH or LiAlH.sub.4), and
others. The coupling agent can also be a magnesium based coupling
agent such as, but not limited to, MgCl.sub.2, iPrMgCl, tBuMgCl,
PhMgCl, or combinations thereof. In some embodiments, the coupling
agent can be a lithium coupling agent or a magnesium coupling
agent. In some embodiments, the coupling agent can be n-BuLi,
MgCl.sub.2, iPrMgCl, tBuMgCl, PhMgCl, or combinations thereof. In
some embodiments, the coupling agent can be n-BuLi. In some
embodiments, the coupling agent can be PhMgCl and iPrMgCl.
[0081] The coupling agent can be present in any suitable amount.
For example, the coupling agent can be present in an amount of at
least 1.0 eq. (mol/mol) to the compound of Formula V, such as about
1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
coupling agent can also be present in an amount of from about 1.0
to about 10.0 eq. (mol/mol) to the compound of Formula V, such as
of from about 1.0 to about 5.0 eq. (mol/mol), or of from about 1.0
to about 2.0 eq. (mol/mol). In some embodiments, the coupling agent
can be present in an amount of from about 1.0 to about 5.0 eq.
(mol/mol) to the compound of Formula V. In some embodiments, the
coupling agent can be present in an amount of from about 1.0 to
about 2.0 eq. (mol/mol) to the compound of Formula V.
[0082] Any suitable halo-silane can be used in the method of making
the compound of Formula V. For example, the halo-silane can be a
fluoro-silane, a chloro-silane, a bromo-silane or an iodo-silane.
The silane portion can have any suitable substituents, such as
alkyl, alkenyl, alkynyl, cycloalkyl, or phenyl. Exemplary
halo-silanes include, but are not limited to,
Cl--Si(CH.sub.3).sub.3, or
Cl--Si(CH.sub.3).sub.2CH.sub.2CH.sub.2Si(CH.sub.3).sub.2--Cl. In
some embodiments, the halo-silane can be a chloro-silane. In some
embodiments, the halo-silane can be Cl--Si(CH.sub.3).sub.3, or
Cl--Si(CH.sub.3).sub.2CH.sub.2CH.sub.2Si(CH.sub.3).sub.2--Cl. In
some embodiments, the halo-silane can be TMSCl.
[0083] The silyl group of R.sup.10 can be any suitable group, but
can depend on the choice of the halo-silane. For example, when the
halo-silane is TMSCl, the silyl group can be trimethylsilyl.
[0084] The halo-silane can be present in any suitable amount. For
example, the halo-silane can be present in an amount of at least
1.0 eq. (mol/mol) to the compound of Formula V, such as about 1.0,
2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
halo-silane can also be present in an amount of from about 1.0 to
about 10.0 eq. (mol/mol) to the compound of Formula V, such as of
from about 1.0 to about 5.0 eq. (mol/mol), or of from about 1.0 to
about 2.0 eq. (mol/mol). In some embodiments, the halo-silane can
be present in an amount of from about 1.0 to about 5.0 eq.
(mol/mol) to the compound of Formula V. In some embodiments, the
halo-silane can be present in an amount of from about 1.0 to about
2.0 eq. (mol/mol) to the compound of Formula V.
[0085] The hydroxy protecting group can be any protecting group
suitable for a hydroxy functional group. Representative hydroxy
protecting groups include, but are not limited to, silanes such as
trimethyl silane (TMS), t-butyl dimethyl silane (TBDMS), or t-butyl
diphenyl silane (TBDPS), ethers such as methyl-methoxy (MOM),
tetrahydropyran (THP), t-butyl, allyl, or benzyl, and esters such
as acetyl, pivaloyl, or benzoyl. In some embodiments, the hydroxy
protecting group can be trimethyl silane (TMS), t-butyl dimethyl
silane (TBDMS), t-butyl diphenyl silane (TBDPS), methyl-methoxy
(MOM), tetrahydropyran (THP), t-butyl, allyl, benzyl, acetyl,
pivaloyl, or benzoyl. In some embodiments, the hydroxy protecting
group can be benzyl.
[0086] Hydroxy groups on adjacent carbons, referred to as
1,2-hydroxy groups, can form a cyclic protecting group called an
acetonide by reaction with a ketone of di-ether. Exemplary
acetonides include, but are not limited to acetonide and
benzylidene acetal. In some embodiments, the hydroxy protecting
groups of hydroxy groups on adjacent carbons can be combined to
form acetonide.
[0087] When the R.sup.19 group is C.sub.1-C.sub.8 alkyl, R.sup.19
can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, the R.sup.19 group
can be methyl.
[0088] Any suitable solvent can be used in the method of the
present invention. Representative solvents include, but are not
limited to, pentane, pentanes, hexane, hexanes, heptane, heptanes,
petroleum ether, cyclopentanes, cyclohexanes, benzene, toluene,
xylene, trifluoromethylbenzene, halobenzenes such as chlorobenzene,
fluorobenzene, dichlorobenzene and difluorobenzene, methylene
chloride, chloroform, acetone, ethyl acetate, diethyl ether,
tetrahydrofuran, or combinations thereof. In some embodiments, the
solvent can be tetrahydrofuran.
[0089] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -78.degree. C. to about 100.degree. C., or of
from about -50.degree. C. to about 100.degree. C., or of from about
-25.degree. C. to about 50.degree. C., or of from about -10.degree.
C. to about 25.degree. C., or of from about 0.degree. C. to about
20.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about 0.degree. C. to about 20.degree.
C.
[0090] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0091] The method of the present invention can provide the compound
of Formula V in any suitable yield. For example, the compound of
Formula V can be prepared in a yield of at least about 50%, 55, 60,
65, 70, 75, 80, 85, 90 or at least about 95%.
[0092] The method of the present invention can provide the compound
of Formula V in any suitable purity. For example, the compound of
Formula V can be prepared in a purity of at least about 90, 95, 96,
97, 98 or at least about 99%. In some embodiments, the compound of
Formula V can be prepared in at least 95% purity. In some
embodiments, the compound of Formula V can be prepared in at least
98% purity. In some embodiments, the compound of Formula V can be
prepared in at least 99% purity.
[0093] In some embodiments, the method including preparing the
compound of Formula V:
##STR00030##
wherein the method includes forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl, the compound of Formula VI:
##STR00031##
and the compound of Formula VII:
##STR00032##
under conditions suitable to prepare the compound of Formula V.
[0094] In some embodiments, the present invention provides the
compound:
##STR00033##
[0095] In some embodiments, the present invention provides a method
of preparing a compound of Formula V-a or V-b:
##STR00034##
The method of making the compound of Formula V-a or Formula V-b
comprises forming a reaction mixture having a deprotonating agent,
a silylating agent, a coupling agent, an additive, a compound of
Formula VI-a:
##STR00035##
and a compound of Formula VII:
##STR00036##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b, wherein each R.sup.b is independently a hydroxy
protecting group, alternatively, two R.sup.b groups on adjacent
carbons can be combined to form a --C(R.sup.19).sub.2-- group,
R.sup.10 is H or a silyl group, and R.sup.19 is H, C.sub.1-C.sub.8
alkyl, phenyl or substituted phenyl.
[0096] Any suitable deprotonating agent can be used in the method
of making the compound of Formula V-a or Formula V-b. The
deprotonating agent can be a sodium deprotonating agent, a
magnesium based deprotonating agent, lithium based deprotonating
agent, potassium based deprotonating agent, or others. For example,
the deprotonating agent can be sodium hydride (NaH),
isopropylmagnesium chloride (iPrMgCl), tert-butylmagnesium chloride
(tBuMgCl), phenylmagnesium chloride (PhMgCl), phenylmagnesium
bromide (PhMgBr), butyllithium (BuLi), methyllithium (MeLi),
methylmagnesium chloride (MeMgCl), methylmagnesium bromide
(MeMgBr), tert-butyllithium (tBuLi), isopropyllithium (iPrLi),
phenyllithium (PhLi), lithium hydride (LiH), potassium hydride
(KH), ethyllithium (EtLi), ethylmagnesium bromide (EtMgBr),
ethylmagnesium chloride (EtMgCl), propyllithium (PrLi),
propylmagnesium bromide (PrMgBr), propylmagnesium chloride
(PrMgCl), cyclohexanelithium (cyHexLi), cyclohexanemagnesium
bromide (cyHexMgBr), cyclohexanemagnesium chloride (cyHexMgCl), or
combinations thereof. In some embodiments, the deprotonating agent
can be PhMgCl.
[0097] The deprotonating agent can be present in any suitable
amount. For example, the deprotonating agent can be present in an
amount of at least 0.1 eq. (mol/mol) to the compound of Formula
VII, such as about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about
10.0 eq. (mol/mol). The deprotonating agent can also be present in
an amount of from about 0.1 to about 10.0 eq. (mol/mol) to the
compound of Formula VII, such as of from about 0.1 to about 3.0 eq.
(mol/mol), or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the deprotonating agent can be present in an amount
from about 0.1 to 1.0 eq. (mol/mol) to the compound of Formula VII.
In some embodiments, the deprotonating agent can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula VII.
[0098] Any suitable silylating agent can be used in the method of
making the compound of Formula V-a or Formula V-b. For example, the
silylating agent can be a fluoro-silane, a chloro-silane, a
bromo-silane or an iodo-silane. For example, the silylating agent
can be a tri-substituted silyl chloride, a tri-substituted silyl
bromide, a tri-substituted silyl iodide, or a tri-substituted silyl
fluoride. The silyl portion can have any suitable substituents,
such as alkyl, alkenyl, alkynyl, cycloalkyl, or phenyl. Exemplary
silylating agents include, but are not limited to,
Cl--Si(CH.sub.3).sub.3,
Cl--Si(CH.sub.3).sub.2CH.sub.2CH.sub.2Si(CH.sub.3).sub.2--Cl, or
tert-butyldiphenylsilyl (TBDPS). In some embodiments, the
silylating agent can be a chloro-silane. In some embodiments, the
silylating agent can be Cl--Si(CH.sub.3).sub.3, or
Cl--Si(CH.sub.3).sub.2CH.sub.2CH.sub.2Si(CH.sub.3).sub.2--Cl. In
some embodiments, the silylating agent can be TMSCl.
[0099] The silyl group of R.sup.10 can be any suitable group, but
can depend on the choice of the silylating agent. For example, when
the silylating agent is TMSCl, the silyl group can be
trimethylsilyl.
[0100] The silylating agent can be present in any suitable amount.
For example, the silylating agent can be present in an amount of at
least 0.0 eq. (mol/mol) to the compound of Formula VII, such as
about 0.0, 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The silylating agent can also be present in an amount of
from about 0.0 to about 10.0 eq. (mol/mol) to the compound of
Formula VII, such as of from about 0.0 to about 3.0 eq. (mol/mol),
or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the silylating agent can be present in an amount from
about 0.0 to 1.0 eq. (mol/mol) to the compound of Formula VII. In
some embodiments, the silylating agent can be present in an amount
of from about 1.0 to about 2.0 eq. (mol/mol) to the compound of
Formula VII.
[0101] Any suitable coupling agent can be used in the method of
making the compound of Formula V-a or Formula V-b. The coupling
agent can be a lithium coupling agent, a magnesium based
deprotonating agent, or others. For example, the coupling agent can
be n-butyllithium (nBuLi), magnesium chloride (MgCl.sub.2),
isopropylmagnesium chloride (iPrMgCl), isopropylmagnesium
chloride-lithium chloride (iPrMgCl--LiCl), tert-butylmagnesium
chloride (tBuMgCl), phenylmagnesium chloride (PhMgCl),
methyllithium (MeLi), methylmagnesium chloride (MeMgCl),
methylmagnesium bromide (MeMgBr), tert-butyllithium (tBuLi),
isopropyllithium (iPrLi), phenyllithium (PhLi), lithium hydride
(LiH), potassium hydride (KH), sodium hydride (NaH), ethyllithium
(EtLi), ethylmagnesium bromide (EtMgBr), ethylmagnesium chloride
(EtMgCl), propyllithium (PrLi), propylmagnesium bromide (PrMgBr),
propylmagnesium chloride (PrMgCl), cyclohexanelithium (cyHexLi),
cyclohexanemagnesium bromide (cyHexMgBr), cyclohexanemagnesium
chloride (cyHexMgCl), or combinations thereof. In some embodiments,
the coupling agent can be iPrMgCl.
[0102] The coupling agent can be present in any suitable amount.
For example, the coupling agent can be present in an amount of at
least 0.1 eq. (mol/mol) to the compound of Formula VII, such as
about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The coupling agent can also be present in an amount of
from about 0.1 to about 10.0 eq. (mol/mol) to the compound of
Formula VII, such as of from about 0.1 to about 3.0 eq. (mol/mol),
or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the coupling agent can be present in an amount from
about 0.1 to 1.0 eq. (mol/mol) to the compound of Formula VII. In
some embodiments, the coupling agent can be present in an amount of
from about 1.0 to about 2.0 eq. (mol/mol) to the compound of
Formula VII.
[0103] Any suitable additive can be used in the method of making
the compound of Formula V-a of Formula V-b. In some embodiments,
the additive is a Lewis Acid. In some embodiments, the additive can
be BF.sub.3-OEt.sub.2, SmOTf).sub.3, Sc(OTf).sub.3, FeCl.sub.3,
LiCl, LiBr, TiCl(OiPr).sub.3, ScCl.sub.3,
Bu.sub.4NBr+LaCl.sub.3-2LiCl, nLaCl.sub.3+mLiCl, wherein m is 0.5
to 50, n is 1 to 100, LaCl.sub.3+2LiCl, Sm(OTf).sub.3+LiCl,
SmCl.sub.3, Bis[2-(N,N-dimethylamino)ethyl]ether, TMEDA,
NdCl.sub.3, NdCl.sub.3+CsCl, nNdCl.sub.3+mLiCl, wherein m is 0.5 to
50, n is 1 to 100, NdCl.sub.3+2LiCl, NdCl.sub.3+LiBr,
NdCl.sub.3+LiI, NdBr.sub.3, NdBr.sub.3+CsCl, nNdBr.sub.3+mLiCl,
wherein m is 0.5 to 50, n is 1 to 100, NdBr.sub.3+2LiCl,
NdBr.sub.3+LiBr, NdBr.sub.3+LiI, Nd(OTf).sub.3, CeCl.sub.3,
CeCl.sub.3+CsCl, nCeCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1
to 100, CeCl.sub.3+2LiCl, CeCl.sub.3+LiBr, CeCl.sub.3+LiI,
CeBr.sub.3, Ce(OTf).sub.3, YCl.sub.3, YCl.sub.3+CsCl,
nYCl.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to 100,
YCl.sub.3+2LiCl, YCl.sub.3+LiBr, YCl.sub.3+LiI, YBr.sub.3,
YBr.sub.3+CsCl, nYBr.sub.3+mLiCl, wherein m is 0.5 to 50, n is 1 to
100, YBr.sub.3+2LiCl, YBr.sub.3+LiBr, YBr.sub.3+LiI, Y(OTf).sub.3,
LaCl.sub.3, La(OTf).sub.3, MgCl.sub.2, TiCl.sub.4, SnCl.sub.4,
AlCl.sub.3, Bu.sub.4NCl, Diethyleneglycol diethylether (DGDE),
DGDE+Bu.sub.4NCl, DGDE+Bu.sub.4NBr, DGDE+Bu.sub.4NI, CaCl.sub.2,
CaBr.sub.2, CaI.sub.2, Ca(OTf).sub.2, YCl.sub.3, YCl.sub.3-2LiCl,
YCl.sub.3--LiCl or a combination thereof. In some embodiments, the
additive can be LiCl, Ca(OTf).sub.2, CaCl.sub.2 and MgCl.sub.2,
CeCl.sub.3, LaCl.sub.3, or a combination thereof. In some
embodiments, the additive can be YCl.sub.3, CeCl.sub.3, NdCl.sub.3,
LaCl.sub.3, or a combination thereof.
[0104] The additive can be present in any suitable amount. For
example, the additive can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula VII, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
additive can also be present in an amount of from about 0.1 to
about 10.0 eq. (mol/mol) to the compound of Formula VII, such as of
from about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0 to
about 2.0 eq. (mol/mol). In some embodiments, the additive can be
present in an amount from about 0.1 to 1.0 eq. (mol/mol) to the
compound of Formula VII. In some embodiments, the additive can be
present in an amount of from about 1.0 to about 2.0 eq. (mol/mol)
to the compound of Formula VII.
[0105] In some embodiments, the additive is LaCl.sub.3-2LiCl and is
present in an amount of at least 0.1 eq. (mol/mol) to the compound
of Formula VII, such as about 0, 0.1, 0.3, 0.5, 1.0, 2, or about
2.0 eq. (mol/mol). In some embodiments, the additive is
LaCl.sub.3-2LiCl and is present in an amount of from about 0 to
about 2.0 eq. (mol/mol) to the compound of Formula VII, such as of
from about 0 to about 0.3 eq. (mol/mol), or of from about 0 to
about 0.5 eq. (mol/mol). In some embodiments, the additive is
LaCl.sub.3-2LiCl and is present in an amount from about 0 to 0.5
eq. (mol/mol) to the compound of Formula VII. In some embodiments,
the the additive is LaCl.sub.3-2LiCl and is present in an amount of
about 0.5 eq. (mol/mol) to the compound of Formula VII.
[0106] In some embodiments, the additive is CeCl.sub.3 and is
present in an amount of at least 0.1 eq. (mol/mol) to the compound
of Formula VII, such as about 0, 0.1, 0.3, 0.5, 1.0, 2, or about
2.0 eq. (mol/mol). In some embodiments, the additive is CeCl.sub.3
and is present in an amount of from about 0 to about 2.0 eq.
(mol/mol) to the compound of Formula VII, such as of from about 0
to about 0.3 eq. (mol/mol), or of from about 0 to about 0.5 eq.
(mol/mol). In some embodiments, the additive is CeCl.sub.3 and is
present in an amount from about 0 to 0.5 eq. (mol/mol) to the
compound of Formula VII. In some embodiments, the the additive is
CeCl.sub.3 and is present in an amount of about 0.5 eq. (mol/mol)
to the compound of Formula VII.
[0107] In some embodiments, the additive is NdCl.sub.3 and is
present in an amount of at least 0.1 eq. (mol/mol) to the compound
of Formula VII, such as about 0, 0.1, 0.3, 0.5, 1.0, 2, or about
2.0 eq. (mol/mol). In some embodiments, the additive is NdCl.sub.3
and is present in an amount of from about 0 to about 2.0 eq.
(mol/mol) to the compound of Formula VII, such as of from about 0
to about 0.3 eq. (mol/mol), or of from about 0 to about 0.5 eq.
(mol/mol). In some embodiments, the additive is NdCl.sub.3 and is
present in an amount from about 0 to 0.5 eq. (mol/mol) to the
compound of Formula VII. In some embodiments, the the additive is
NdCl.sub.3 and is present in an amount of about 0.5 eq. (mol/mol)
to the compound of Formula VII.
[0108] In some embodiments, the additive is YCl.sub.3 and is
present in an amount of at least 0.1 eq. (mol/mol) to the compound
of Formula VII, such as about 0, 0.1, 0.3, 0.5, 1.0, 2, or about
2.0 eq. (mol/mol). In some embodiments, the additive is YCl.sub.3
and is present in an amount of from about 0 to about 2.0 eq.
(mol/mol) to the compound of Formula VII, such as of from about 0
to about 0.3 eq. (mol/mol), or of from about 0 to about 0.5 eq.
(mol/mol). In some embodiments, the additive is YCl.sub.3 and is
present in an amount from about 0 to 0.5 eq. (mol/mol) to the
compound of Formula VII. In some embodiments, the the additive is
YCl.sub.3 and is present in an amount of about 0.5 eq. (mol/mol) to
the compound of Formula VII.
[0109] When the R.sup.19 group is C.sub.1-C.sub.8 alkyl, R.sup.19
can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, the R.sup.19 group
can be methyl.
[0110] When the R.sup.b group is a hydroxy protecting group,
R.sup.b can be any example protecting group described in Protective
Groups in Organic Chemistry, Peter G. M. Wuts and Theodora W.
Greene, 4th Ed., 2006. In some embodiments, the R.sup.b group can
be benzyl. In some embodiments, the R.sup.b group can be TBS.
[0111] The hydroxy protecting group can be any protecting group
suitable for a hydroxy functional group. Representative hydroxy
protecting groups include, but are not limited to, silanes, ethers,
esters, or others. Representative hydroxy protecting groups
include, but are not limited to trimethyl silane (TMS), t-butyl
dimethyl silane (TBDMS), t-butyl diphenyl silane (TBDPS),
methyl-methoxy (MOM), tetrahydropyran (THP), t-butyl, allyl,
benzyl, acetyl, pivaloyl, or benzoyl. In some embodiments, the
hydroxy protecting group can be trimethyl silane (TMS), t-butyl
dimethyl silane (TBDMS), t-butyl diphenyl silane (TBDPS),
methyl-methoxy (MOM), tetrahydropyran (THP), t-butyl, allyl,
benzyl, acetyl, pivaloyl, or benzoyl. In some embodiments, the
hydroxy protecting group can be benzyl. In some embodiments, the
hydroxy protecting group can be TBS.
[0112] Hydroxy groups on adjacent carbons, referred to as
1,2-hydroxy groups, can form a cyclic protecting group called an
acetal or a ketal by reaction with an aldehyde, an acetale,
aketoneor a ketal. Exemplary acetals and ketals include, but are
not limited to a, benzylidene acetal and an acetonide. In some
embodiments, the hydroxy protecting groups of hydroxy groups on
adjacent carbons can be combined to form acetonide.
[0113] Any suitable solvent can be used in the method of the
present invention. Representative solvents include, but are not
limited to, pentane, pentanes, hexane, hexanes, heptane, heptanes,
petroleum ether, cyclopentanes, cyclohexanes, benzene, toluene,
xylene, dichloromethane, trifluoromethylbenzene, halobenzenes such
as chlorobenzene, fluorobenzene, dichlorobenzene and
difluorobenzene, methylene chloride, chloroform, acetone, ethyl
acetate, diethyl ether, tetrahydrofuran (THF),
2-methyltetrahydrofuran, dibutyl ether, diisopropyl ether, methyl
tert-butyl ether, dimethoxyethane, dioxanes (1.4 dioxane), N-methyl
pyrrolidinone (NMP), diisopropyl ether, or combinations thereof. In
certain embodiments, the solvent can be THF, MeTHF, toluene,
THF+dioxane, THF+pyridine, or THF+DCM, or combinations thereof. In
some embodiments, the solvent can be THF.
[0114] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be from about -78.degree. C. to about 100.degree. C., or from
about -50.degree. C. to about 100.degree. C., or from about
-25.degree. C. to about 50.degree. C., or from about -10.degree. C.
to about 25.degree. C., or from about 0.degree. C. to about
20.degree. C. In some embodiments, the temperature of the reaction
mixture can be from about 0.degree. C. to about 20.degree. C. In
some embodiments, the temperature of the reaction mixture can be
from about -30.degree. C. to about -10.degree. C.
[0115] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0116] The method of the present invention can provide the compound
of Formula V-a or Formula V-b in any suitable yield. For example,
the compound of Formula V-a or Formula V-b can be prepared in a
yield of at least about 50%, 55, 60, 65, 70, 75, 80, 85, 90 or at
least about 95%.
[0117] The method of the present invention can provide the compound
of Formula V-a or Formula V-b in any suitable purity. For example,
the compound of Formula V-a or Formula V-b can be prepared in a
purity of at least about 90, 95, 96, 97, 98 or at least about 99%.
In some embodiments, the compound of Formula V-a or Formula V-b can
be prepared in at least 95% purity. In some embodiments, the
compound of Formula V-a or Formula V-b can be prepared in at least
98% purity. In some embodiments, the compound of Formula V-a or
Formula V-b can be prepared in at least 99% purity.
[0118] In some embodiments, the method comprises preparing the
compound of Formula V-a or Formula V-b:
##STR00037##
wherein the method comprises forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl, LaCl.sub.3-2LiCl the compound of Formula
VI:
##STR00038##
and the compound of Formula VII:
##STR00039##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b.
[0119] In some embodiments, the method comprises preparing the
compound of Formula V-a or Formula V-b:
##STR00040##
wherein the method comprises forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl, CeCl.sub.3 the compound of Formula VI:
##STR00041##
and the compound of Formula VII:
##STR00042##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b.
[0120] In some embodiments, the method comprises preparing the
compound of Formula V-a or Formula V-b:
##STR00043##
wherein the method comprises forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl, NdCl.sub.3 the compound of Formula VI:
##STR00044##
and the compound of Formula VII:
##STR00045##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b.
[0121] In some embodiments, the method comprises preparing the
compound of Formula V-a or Formula V-b:
##STR00046##
wherein the method comprises forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl, YCl.sub.3 the compound of Formula VI:
##STR00047##
and the compound of Formula VII:
##STR00048##
under conditions suitable to prepare the compound of Formula V-a or
Formula V-b.
[0122] In some embodiments, the method comprises preparing the
compound of Formula V-a:
##STR00049##
wherein the method comprises forming the reaction mixture having
TMSCl, PhMgCl, iPrMgCl--LiCl, LaCl.sub.3-2LiCl the compound of
Formula VI:
##STR00050##
and the compound of Formula VII:
##STR00051##
under conditions suitable to prepare the compound of Formula
V-a.
[0123] B. Preparation of Cyano Nucleosides
[0124] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI:
##STR00052##
wherein R.sup.c is H or a hydroxy protecting group, or two R.sup.c
on adjacent carbons can be combined to form a --C(R.sup.19).sub.2--
group, and R.sup.19 is H or C.sub.1-C.sub.8 alkyl.
[0125] When the R.sup.19 group is C.sub.1-C.sub.8 alkyl, R.sup.19
can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, the R.sup.19 group
can be methyl.
[0126] When the R.sup.c group is a hydroxy protecting group, the
hydroxy protecting group can be any protecting group suitable for a
hydroxy functional group. Representative hydroxy protecting groups
include, but are not limited to, silanes, ethers, esters, or
others. Representative hydroxy protecting groups include, but are
not limited to trimethyl silane (TMS), t-butyl dimethyl silane
(TBDMS), t-butyl diphenyl silane (TBDPS), methyl-methoxy (MOM),
tetrahydropyran (THP), t-butyl, allyl, benzyl, acetyl, pivaloyl, or
benzoyl. In some embodiments, the hydroxy protecting group can be
trimethyl silane (TMS), t-butyl dimethyl silane (TBDMS), t-butyl
diphenyl silane (TBDPS), methyl-methoxy (MOM), tetrahydropyran
(THP), t-butyl, allyl, benzyl, acetyl, pivaloyl, or benzoyl. In
some embodiments, the hydroxy protecting group can be benzyl. In
some embodiments, the hydroxy protecting group can be TBS
[0127] Hydroxy groups on adjacent carbons, referred to as
1,2-hydroxy groups, can form a cyclic protecting group called an
acetal or a ketal by reaction with an aldehyde, an acetale,
aketoneor a ketal. Exemplary acetals and ketals include, but are
not limited to a, benzylidene acetal and an acetonide. In some
embodiments, the hydroxy protecting groups of hydroxy groups on
adjacent carbons can be combined to form acetonide.
[0128] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-a:
##STR00053##
wherein the method comprises forming a reaction mixture having a
cyanating agent, a Lewis Acid, a Broenstedt acid, a solvent, and
the compound of Formula V-a or V-b:
##STR00054##
under conditions suitable to prepare the compound of Formula XI-a,
wherein R.sup.b is independently a hydroxy protecting group,
alternatively, two R.sup.b groups on adjacent carbons can be
combined to form a --C(R.sup.19).sub.2 group, R.sup.19 is H or a
silyl group, and R.sup.19 is H, C.sub.1-C.sub.8 alkyl, phenyl or
substituted phenyl.
[0129] Any suitable cyanating agent can be used in the method of
making the compound of Formula XI-a. For example, the cyanating
agent can be TMSCN, TBSCN, TESCN, HCN, KCN, NaCN, 4-toluenesulfonyl
cyanide, CuCN, CuCn*LiCl, LiCN, Zn(CN).sub.2,
K.sub.4[Fe(CN).sub.6], tetrabutylammonium cyanide,
tetrmethylammonium cyanide, tetraethylammonium cyanide,
tetrabutylammonium cyanide, (including tetraalkylammonium cyanide
with alkyl independently being Me, Et, Pr, iPr, Bu, iBu, tertBu,
Pent, Hex), tributyltn cyanide, trimethyltin cyanide, triethyltin
cyanide, tripropyltin cyanide, (including trialkyltin cyanide
cyanide with alkyl independently being Me, Et, Pr, iPr, Bu, iBu,
tertBu, Pent, Hex), 2-hydroxy-2-methylpropanenitrile; or
combinations thereof. In some embodiments, the cyanating agent can
be TMSCN.
[0130] The cyanating agent can be present in any suitable amount.
For example, the cyanating agent can be present in an amount of at
least 0.1 eq. (mol/mol) to the compound of Formula V-a or Formula
V-b, such as about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about
10.0 eq. (mol/mol). The cyanating agent can also be present in an
amount of from about 0.1 to about 10.0 eq. (mol/mol) to the
compound of Formula V-a or Formula V-b, such as of from about 0.1
to about 3.0 eq. (mol/mol), or of from about 1.0 to about 2.0 eq.
(mol/mol). In some embodiments, the cyanating agent can be present
in an amount from about 0.1 to 1.0 eq. (mol/mol) to the compound of
Formula V-a or Formula V-b. In some embodiments, the cyanating
agent can be present in an amount of from about 1.0 to about 2.0
eq. (mol/mol) to the compound of Formula V-a or Formula V-b.
[0131] Any suitable Lewis Acid can be used in the method of making
the compound of Formula XI-a. For example, the Lewis Acid can be
TMSOTf, TMSOTf, TBSOTf, TESOTf, BF.sub.3, BF.sub.3-OEt.sub.2,
BCl.sub.3, BF.sub.3-THF, MgCl.sub.2, MgI.sub.2, MgBr.sub.2,
MgBr.sub.2-OEt.sub.2, ZnCl.sub.2, ZnBr.sub.2, ZnI.sub.2, LiCl,
LiBr, LiI, AlCl.sub.3, AlBr.sub.3, AlI.sub.3,
Me.sub.2Si(OTf).sub.2, Et.sub.2Si(OTf).sub.2,
Pr.sub.2Si(OTf).sub.2, iPr.sub.2Si(OTf).sub.2,
(tBu).sub.2Si(OTf).sub.2, (C.sub.6F.sub.5).sub.3B, MeSiCl.sub.3,
Me.sub.2SiCl.sub.2, SiCl.sub.4, TMSCl, TMSI, TMSVr, TBSCl, TBSBr,
TBSI, TESCl, TESBr, TESI, SmCl.sub.3, SmBr.sub.3, SmI.sub.2,
SmI.sub.3, ScI.sub.3, ScBr.sub.3, ScI.sub.3, Sm(OTf).sub.3,
Sc(OTf).sub.3, TiCl.sub.4, Ti(OiPr).sub.4, Ti(OiPr).sub.3Cl,
Ti(OiPr).sub.2Cl.sub.2, Ti(OiPr)Cl.sub.3, Zn(BF.sub.4).sub.2,
LiBF.sub.4, Mg(BF.sub.4).sub.2, ZrCl.sub.4, FeCl.sub.2, FeCl.sub.3,
FeBr.sub.2, FeBr.sub.3, FeI.sub.2, FeI.sub.3, Cu(OTf),
Cu(OTf).sub.2, 4-toluenesulfonylchoride, benzenesulfonylchlopride,
4-toluenesulfonyl triflate, benzenesulfonyl triflate,
methylsulfonyl chloride, methylsulfonic anhydrate, InCl.sub.3,
InBr.sub.3, InI.sub.3, In(OTf).sub.3, Mg(SO.sub.4).sub.2,
NaSO.sub.4; or combinations thereof. In some embodiments, the Lewis
Acid can be TMSOTf. In some embodiments, the following may be used
in the method of making the compound of Formula XI-a instead of a
Lewis Acid: dicyclohexylcarbodiimide,
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, benzenesulfonic
acid, HCl, 4-toluenesulfonic acid, triflic acid, trifluoroacetic
acid, 4-nitrobenzolic acid, methylsoulfonic acid, sulfuric acid,
phosphoric acid, HBr, acetic acid, formic acid, HI; or combinations
thereof.
[0132] The Lewis Acid can be present in any suitable amount. For
example, the Lewis Acid can be present in an amount of at least 0.0
eq. (mol/mol) to the compound of Formula V-a or Formula V-b, such
as about 0.0, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The Lewis Acid can also be present in an amount of from
about 0.0 to about 10.0 eq. (mol/mol) to the compound of Formula
V-a or Formula V-b, such as of from about 0.0 to about 3.0 eq.
(mol/mol), or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the Lewis Acid can be present in an amount from about
0.0 to 1.0 eq. (mol/mol) to the compound of Formula V-a or Formula
V-b. In some embodiments, the Lewis Acid can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula V-a or Formula V-b.
[0133] Any suitable Broenstedt acid can be used in the method of
making the compound of Formula XI-a. For example, the Broenstedt
acid can be TFA, benzenesulfonic acid, HCl, 4-toluenesulfonic acid,
triflic acid, trifluoroacetic acid, 4-nitrobenzoic acid,
methylsoulfonic acid, sulfuric acid, phosphoric acid, HBr, acetic
acid, formic acid, HI, trifluoromethylsulfonic acid,
4-fluorobenzoic acid, pivalic acid, HBF.sub.4, nitric acid,
4-chloro-benzoic acid, pentafluorophenol, HPF.sub.6,
Camphorsulfonic acid; or combinations thereof. In some embodiments,
the Broenstedt acid can be TFA.
[0134] The Broenstedt acid can be present in any suitable amount.
For example, the Broenstedt acid can be present in an amount of at
least about 0.0 eq. (mol/mol) to the compound of Formula V-a or
Formula V-b, such as about 0.0, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9,
or about 10.0 eq. (mol/mol). The Broenstedt acid can also be
present in an amount of from about 0.0 to about 10.0 eq. (mol/mol)
to the compound of Formula V-a or Formula V-b, such as of from
about 0.0 to about 3.0 eq. (mol/mol), or of from about 1.0 to about
2.0 eq. (mol/mol). In some embodiments, the Broenstedt acid can be
present in an amount from about 0.0 to about 1.0 eq. (mol/mol) to
the compound of Formula V-a or Formula V-b. In some embodiments,
the Broenstedt acid can be present in an amount of from about 1.0
to about 2.0 eq. (mol/mol) to the compound of Formula V-a or
Formula V-b.
[0135] Any suitable solvent can be used in the method of making the
compound of Formula XI-a. For example, the solvent can be DCM, THF,
MeTHF, Et.sub.2O, MeCN, EtCN, toluene, benzene, chlorobenzene,
nitrobenzene, flurorbenzene, methanol, ethanol, 2-propanol,
propanol, butanol, MTBE, EtOAc, iPrOAc, Me2O, (TMS)2O, acetone,
2-butanone, chloroform, 1,2-dichloroethane, diglyme, dioxane,
acetic acid, formic acid, trifluoroacetic acid,
methylisobutylketone, DMAc, DMF, NMP, DMSO; or combinations
thereof. In some embodiments, the solvent can be DCM.
[0136] The solvent can be present in any suitable amount. For
example, the solvent can be present in an amount of at least 0.0
eq. (mol/mol) to the compound of Formula V-a or Formula V-b, such
as about 0.0, 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0
eq. (mol/mol). The solvent can also be present in an amount of from
about 0.0 to about 10.0 eq. (mol/mol) to the compound of Formula
V-a or Formula V-b, such as of from about 0.0 to about 3.0 eq.
(mol/mol), or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the solvent can be present in an amount from about 0.1
to about 1.0 eq. (mol/mol) to the compound of Formula V-a or
Formula V-b. In some embodiments, the solvent can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula V-a or Formula V-b.
[0137] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -150.degree. C. to about 0.degree. C., or of
from about -120.degree. C. to about 0.degree. C., or of from about
-100.degree. C. to about 0.degree. C., or of from about
-100.degree. C. to about -50.degree. C., or of from about
-100.degree. C. to about -70.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-120.degree. C. to about -70.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-120.degree. C. to about -100.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-80.degree. C. to about -30.degree. C.
[0138] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0139] The method of the present invention can provide the compound
of Formula XI-a in any suitable yield. For example, the compound of
Formula XI-a can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0140] The method of the present invention can provide the compound
of Formula XI-a in any suitable purity. For example, the compound
of Formula XI-a can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula XI-a can be prepared in at least about 95%
purity. In some embodiments, the compound of Formula XI-a can be
prepared in at least about 98% purity. In some embodiments, the
compound of Formula XI-a can be prepared in at least about 99%
purity.
[0141] In some embodiments, the method of the present invention can
be performed as a batch mode process. In some embodiments, the
method of the present invention can be performed as a flow
process.
[0142] In some embodiments, the method comprises preparing the
compound of Formula XI-a:
##STR00055##
wherein the method comprises forming the reaction mixture having
TFA, TMSCN, TMSOTf and the compound of Formula Va or Formula
V-b:
##STR00056##
under conditions suitable to prepare the compound of Formula XI-a.
In certain embodiments, the method of preparing Formula XI-a is
performed between about -120.degree. C. and about 20.degree. C. In
another embodiment, the method of preparing Formula XI-a is
performed between about -120.degree. C. and about 0.degree. C. In
another embodiment, the method of preparing Formula XI-a is
performed between about -40.degree. C. and about -20.degree. C.
[0143] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-a.sup.2:
##STR00057##
wherein the method comprises forming a reaction mixture having a
cyanating agent, a Lewis Acid, a Broenstedt acid, solvent, and the
compound of Formula V-a:
##STR00058##
under conditions suitable to prepare the compound of Formula
XI-a.sup.2, wherein R.sup.b is independently a hydroxy protecting
group, alternatively, two R.sup.b groups on adjacent carbons can be
combined to form a --C(R.sup.19).sub.2 group, R.sup.10 is H or a
silyl group, and R.sup.19 is H, C.sub.1-C.sub.8 alkyl, phenyl or
substituted phenyl.
[0144] Any suitable cyanating agent can be used in the method of
making the compound of Formula XI-a.sup.2. For example, the
cyanating agent can be TMSCN, TBSCN, TESCN, HCN, KCN, NaCN,
4-toluenesulfonyl cyanide, CuCN, CuCn*LiCl, LiCN, Zn(CN).sub.2,
K.sub.4[Fe(CN).sub.6], tetrabutylammonium cyanide,
tetrmethylammonium cyanide, tetraethylammonium cyanide,
tetrabutylammonium cyanide, (including tetraalkylammonium cyanide
with alkyl independently being Me, Et, Pr, iPr, Bu, iBu, tertBu,
Pent, Hex), tributyltn cyanide, trimethyltin cyanide, triethyltin
cyanide, tripropyltin cyanide, (including trialkyltin cyanide
cyanide with alkyl independently being Me, Et, Pr, iPr, Bu, iBu,
tertBu, Pent, Hex), 2-hydroxy-2-methylpropanenitrile; or
combinations thereof. In some embodiments, the cyanating agent can
be TMSCN.
[0145] The cyanating agent can be present in any suitable amount.
For example, the cyanating agent can be present in an amount of at
least 0.1 eq. (mol/mol) to the compound of Formula V-a, such as
about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The cyanating agent can also be present in an amount of
from about 0.1 to about 10.0 eq. (mol/mol) to the compound of
Formula V-a, such as of from about 0.1 to about 3.0 eq. (mol/mol),
or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the cyanating agent can be present in an amount from
about 0.1 to 1.0 eq. (mol/mol) to the compound of Formula V-a. In
some embodiments, the cyanating agent can be present in an amount
of from about 1.0 to about 2.0 eq. (mol/mol) to the compound of
Formula V-a.
[0146] Any suitable Lewis Acid can be used in the method of making
the compound of Formula XI-a.sup.2. For example, the Lewis Acid can
be TMSOTf, TMSOTf, TBSOTf, TESOTf, BF.sub.3, BF.sub.3-OEt.sub.2,
BCl.sub.3, BF.sub.3-THF, MgCl.sub.2, MgI.sub.2, MgBr.sub.2,
MgBr.sub.2-OEt.sub.2, ZnCl.sub.2, ZnBr.sub.2, ZnI.sub.2, LiCl,
LiBr, LiI, AlCl.sub.3, AlBr.sub.3, AlI.sub.3,
Me.sub.2Si(OTf).sub.2, Et.sub.2Si(OTf).sub.2,
Pr.sub.2Si(OTf).sub.2, iPr.sub.2Si(OTf).sub.2,
(tBu).sub.2Si(OTf).sub.2, (C.sub.6F.sub.5).sub.3B, MeSiCl.sub.3,
Me.sub.2SiCl.sub.2, SiCl.sub.4, TMSCl, TMSI, TMSVr, TBSCl, TBSBr,
TBSI, TESCl, TESBr, TESI, SmCl.sub.3, SmBr.sub.3, SmI.sub.2,
SmI.sub.3, ScI.sub.3, ScBr.sub.3, ScI.sub.3, Sm(OTf).sub.3,
Sc(OTf).sub.3, TiCl.sub.4, Ti(OiPr).sub.4, Ti(OiPr).sub.3Cl,
Ti(OiPr).sub.2Cl.sub.2, Ti(OiPr)Cl.sub.3, Zn(BF.sub.4).sub.2,
LiBF.sub.4, Mg(BF.sub.4).sub.2, ZrCl.sub.4, FeCl.sub.2, FeCl.sub.3,
FeBr.sub.2, FeBr.sub.3, FeI.sub.2, FeI.sub.3, Cu(OTf),
Cu(OTf).sub.2, 4-toluenesulfonylchoride, benzenesulfonylchlopride,
4-toluenesulfonyl triflate, benzenesulfonyl triflate,
methylsulfonyl chloride, methylsulfonic anhydrate, InCl.sub.3,
InBr.sub.3, InI.sub.3, In(OTf).sub.3, Mg(SO.sub.4).sub.2,
NaSO.sub.4; or combinations thereof. In some embodiments, the Lewis
Acid can be TMSOTf. In some embodiments, the following may be used
in the method of making the compound of Formula XI-a.sup.2 instead
of a Lewis Acid: dicyclohexylcarbodiimide,
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, benzenesulfonic
acid, HCl, 4-toluenesulfonic acid, triflic acid, trifluoroacetic
acid, 4-nitrobenzolic acid, methylsoulfonic acid, sulfuric acid,
phosphoric acid, HBr, acetic acid, formic acid, HI; or combinations
thereof.
[0147] The Lewis Acid can be present in any suitable amount. For
example, the Lewis Acid can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula V-a, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
Lewis Acid can also be present in an amount of from about 0.1 to
about 10.0 eq. (mol/mol) to the compound of Formula V-a, such as of
from about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0 to
about 2.0 eq. (mol/mol). In some embodiments, the Lewis Acid can be
present in an amount from about 0.1 to 1.0 eq. (mol/mol) to the
compound of Formula V-a. In some embodiments, the Lewis Acid can be
present in an amount of from about 1.0 to about 2.0 eq. (mol/mol)
to the compound of Formula V-a.
[0148] Any suitable Broenstedt acid can be used in the method of
making the compound of Formula XI-a.sup.2. For example, the
Broenstedt acid can be TFA, benzenesulfonic acid, HCl,
4-toluenesulfonic acid, triflic acid, trifluoroacetic acid,
4-nitrobenzoic acid, methylsoulfonic acid, sulfuric acid,
phosphoric acid, HBr, acetic acid, formic acid, HI,
trifluoromethylsulfonic acid, 4-fluorobenzoic acid, pivalic acid,
HBF.sub.4, nitric acid, 4-chloro-benzoic acid, pentafluorophenol,
HPF.sub.6, Camphorsulfonic acid; or combinations thereof. In some
embodiments, the Broenstedt acid can be TFA.
[0149] The Broenstedt acid can be present in any suitable amount.
For example, the Broenstedt acid can be present in an amount of at
least about 0.1 eq. (mol/mol) to the compound of Formula V-a, such
as about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The Broenstedt acid can also be present in an amount of
from about 0.1 to about 10.0 eq. (mol/mol) to the compound of
Formula V-a, such as of from about 0.1 to about 3.0 eq. (mol/mol),
or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the Broenstedt acid can be present in an amount from
about 0.1 to about 1.0 eq. (mol/mol) to the compound of Formula
V-a. In some embodiments, the Broenstedt acid can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula V-a.
[0150] Any suitable solvent can be used in the method of making the
compound of Formula XI or XI-a.sup.2. For example, the solvent can
be DCM, THF, MeTHF, Et.sub.2O, MeCN, EtCN, toluene, benzene,
chlorobenzene, nitrobenzene, flurorbenzene, methanol, ethanol,
2-propanol, propanol, butanol, MTBE, EtOAc, iPrOAc, Me2O, (TMS)2O,
acetone, 2-butanone, chloroform, 1,2-dichloroethane, diglyme,
dioxane, acetic acid, formic acid, trifluoroacetic acid,
methylisobutylketone, DMAc, DMF, NMP, DMSO; or combinations
thereof. In some embodiments, the solvent can be DCM.
[0151] The solvent can be present in any suitable amount. For
example, the solvent can be present in an amount of at least 0.0
eq. (mol/mol) to the compound of Formula V-a, such as about 0.0,
0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol).
The solvent can also be present in an amount of from about 0.0 to
about 10.0 eq. (mol/mol) to the compound of Formula V-a, such as of
from about 0.0 to about 3.0 eq. (mol/mol), or of from about 1.0 to
about 2.0 eq. (mol/mol). In some embodiments, the solvent can be
present in an amount from about 0.1 to about 1.0 eq. (mol/mol) to
the compound of Formula V-a. In some embodiments, the solvent can
be present in an amount of from about 1.0 to about 2.0 eq.
(mol/mol) to the compound of Formula V-a.
[0152] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -150.degree. C. to about 0.degree. C., or of
from about -120.degree. C. to about 0.degree. C., or of from about
-100.degree. C. to about 0.degree. C., or of from about
-100.degree. C. to about -50.degree. C., or of from about
-100.degree. C. to about -70.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-120.degree. C. to about -70.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-120.degree. C. to about -100.degree. C. In some embodiments, the
temperature of the reaction mixture can be of from about
-80.degree. C. to about -30.degree. C.
[0153] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0154] The method of the present invention can provide the compound
of Formula XI-a.sup.2 in any suitable yield. For example, the
compound of Formula XI-a.sup.2 can be prepared in a yield of at
least about 50%, 55, 60, 65, 70, 75, 80, 85, 90 or at least about
95%.
[0155] The method of the present invention can provide the compound
of Formula XI-a.sup.2 in any suitable purity. For example, the
compound of Formula XI-a.sup.2 can be prepared in a purity of at
least about 90, 95, 96, 97, 98 or at least about 99%. In some
embodiments, the compound of Formula XI-a.sup.2 can be prepared in
at least about 95% purity. In some embodiments, the compound of
Formula XI-a.sup.2 can be prepared in at least about 98% purity. In
some embodiments, the compound of Formula XI-a.sup.2 can be
prepared in at least about 99% purity.
[0156] In some embodiments, the method of the present invention can
be performed as a batch mode process. In some embodiments, the
method of the present invention can be performed as a flow
process.
[0157] In some embodiments, the method comprises preparing the
compound of Formula XI-a.sup.2:
##STR00059##
wherein the method comprises forming the reaction mixture having
TFA, TMSCN, TMSOTf and the compound of Formula Va:
##STR00060##
under conditions suitable to prepare the compound of Formula
XI-a.sup.2. In certain embodiments, the method of preparing Formula
XI-a.sup.2 is performed between about -120.degree. C. and about
20.degree. C. In another embodiment, the method of preparing
Formula XI-a.sup.2 is performed between about -120.degree. C. and
about 0.degree. C. In another embodiment, the method of preparing
Formula XI-a.sup.2 is performed between about -40.degree. C. and
about -20.degree. C.
[0158] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-b:
##STR00061##
wherein the method comprises forming a reaction mixture having a
Lewis Acid, a base, a solvent, a filtering agent, and the compound
of Formula XI-a
##STR00062##
under conditions suitable to prepare the compound of Formula
XI-b.
[0159] Any suitable Lewis Acid can be used in the method of making
the compound of Formula XI-b. For example, the Lewis Acid can be
TMSOTf, TMSOTf, TBSOTf, TESOTf, BF.sub.3, BF.sub.3-OEt.sub.2,
BCl.sub.3, BF.sub.3-THF, MgCl.sub.2, MgI.sub.2, MgBr.sub.2,
MgBr.sub.2-OEt.sub.2, ZnCl.sub.2, ZnBr.sub.2, ZnI.sub.2, LiCl,
LiBr, LiI, AlCl.sub.3, AlBr.sub.3, AlI.sub.3,
Me.sub.2Si(OTf).sub.2, Et.sub.2Si(OTf).sub.2,
Pr.sub.2Si(OTf).sub.2, iPr.sub.2Si(OTf).sub.2,
(tBu).sub.2Si(OTf).sub.2, (C.sub.6F.sub.5).sub.3B, MeSiCl.sub.3,
Me.sub.2SiCl.sub.2, SiCl.sub.4, TMSCl, TMSI, TMSVr, TBSCl, TBSBr,
TBSI, TESCl, TESBr, TESI, SmCl.sub.3, SmBr.sub.3, SmI.sub.2,
SmI.sub.3, ScI.sub.3, ScBr.sub.3, ScI.sub.3, Sm(OTf).sub.3,
Sc(OTf).sub.3, TiCl.sub.4, Ti(OiPr).sub.4, Ti(OiPr).sub.3Cl,
Ti(OiPr).sub.2Cl.sub.2, Ti(OiPr)Cl.sub.3, Zn(BF.sub.4).sub.2,
LiBF.sub.4, Mg(BF.sub.4).sub.2, ZrCl.sub.4, FeCl.sub.2, FeCl.sub.3,
FeBr.sub.2, FeBr.sub.3, FeI.sub.2, FeI.sub.3, Cu(OTf),
Cu(OTf).sub.2, 4-toluenesulfonylchoride, benzenesulfonylchlopride,
4-toluenesulfonyl triflate, benzenesulfonyl triflate,
methylsulfonyl chloride, methylsulfonic anhydrate, InCl.sub.3,
InBr.sub.3, InI.sub.3, In(OTf).sub.3, Mg(SO.sub.4).sub.2,
NaSO.sub.4; or combinations thereof. In some embodiments, the Lewis
Acid can be BCL.sub.3. In some embodiments, the following may be
used in the method of making the compound of Formula XI-b instead
of a Lewis Acid: dicyclohexylcarbodiimide,
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, benzenesulfonic
acid, HCl, 4-toluenesulfonic acid, triflic acid, trifluoroacetic
acid, 4-nitrobenzolic acid, methylsoulfonic acid, sulfuric acid,
phosphoric acid, HBr, acetic acid, formic acid, HI; or combinations
thereof.
[0160] The Lewis Acid can be present in any suitable amount. For
example, the Lewis Acid can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula XI-a, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
Lewis Acid can also be present in an amount of from about 0.1 to
about 10.0 eq. (mol/mol) to the compound of Formula XI-a, such as
of from about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0
to about 2.0 eq. (mol/mol). In some embodiments, the Lewis Acid can
be present in an amount from about 0.1 to 1.0 eq. (mol/mol) to the
compound of Formula XI-a. In some embodiments, the Lewis Acid can
be present in an amount of from about 1.0 to about 2.0 eq.
(mol/mol) to the compound of Formula XI-a.
[0161] Any suitable base can be used in the method of making the
compound of Formula XI-b. For example, the base can be
(C.sub.1-8Alkyl).sub.3N. In some embodiments, the base can be
Et.sub.3N.
[0162] The base can be present in any suitable amount. For example,
the base can be present in an amount of at least 0.1 eq. (mol/mol)
to the compound of Formula XI-a, such as about 0.1, 0.5, 1.0, 2, 3,
4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The base can also be
present in an amount of from about 0.1 to about 10.0 eq. (mol/mol)
to the compound of Formula XI-a, such as of from about 0.1 to about
3.0 eq. (mol/mol), or of from about 1.0 to about 2.0 eq. (mol/mol).
In some embodiments, the base can be present in an amount from
about 0.1 to 1.0 eq. (mol/mol) to the compound of Formula XI-a. In
some embodiments, the base can be present in an amount of from
about 1.0 to about 2.0 eq. (mol/mol) to the compound of Formula
XI-a.
[0163] Any suitable solvent can be used in the method of making the
compound of Formula XI-b. For example, the solvent can be MeOH,
DCM, THF, MeTHF, Et.sub.2O, MeCN, EtCN, toluene, benzene,
chlorobenzene, nitrobenzene, flurorbenzene, methanol, ethanol,
2-propanol, propanol, butanol, MTBE, EtOAc, iPrOAc, Me2O, (TMS)2O,
acetone, 2-butanone, chloroform, 1,2-dichloroethane, diglyme,
dioxane, acetic acid, formic acid, trifluoroacetic acid,
methylisobutylketone, DMAc, DMF, NMP, DMSO; or combinations
thereof. In some embodiments, the solvent can be MeOH.
[0164] The solvent can be present in any suitable amount. For
example, the solvent can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula XI-a, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
solvent can also be present in an amount of from about 0.1 to about
10.0 eq. (mol/mol) to the compound of Formula XI-a, such as of from
about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0 to about
2.0 eq. (mol/mol). In some embodiments, the solvent can be present
in an amount from about 0.1 to about 1.0 eq. (mol/mol) to the
compound of Formula XI-a. In some embodiments, the solvent can be
present in an amount of from about 1.0 to about 2.0 eq. (mol/mol)
to the compound of Formula XI-a.
[0165] Any suitable filtering agent can be used in the method of
making the compound of Formula XI-b. For example, the filtering
agent can be silica gel, Celite.RTM. or combinations thereof. In
some embodiments, the filtering agent can be Celite.RTM..
[0166] The filtering agent can be present in any suitable amount.
For example, the filtering agent can be present in an amount of at
least 0.1 eq. (mol/mol) to the compound of Formula XI-a, such as
about 0.1, 0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq.
(mol/mol). The filtering agent can also be present in an amount of
from about 0.1 to about 10.0 eq. (mol/mol) to the compound of
Formula XI-a, such as of from about 0.1 to about 3.0 eq. (mol/mol),
or of from about 1.0 to about 2.0 eq. (mol/mol). In some
embodiments, the filtering agent can be present in an amount from
about 0.1 to about 1.0 eq. (mol/mol) to the compound of Formula
XI-a. In some embodiments, the filtering agent can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula XI-a.
[0167] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -50.degree. C. to about 0.degree. C., or of
from about -40.degree. C. to about 0.degree. C., or of from about
-30.degree. C. to about 0.degree. C., or of from about -20.degree.
C. to about 0.degree. C., or of from about -20.degree. C. to about
-10.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about -30.degree. C. to about 0.degree. C.
In some embodiments, the temperature of the reaction mixture can be
of from about -20.degree. C. to about -10.degree. C. In some
embodiments, the temperature of the reaction mixture can be of from
about -25.degree. C. to about -15.degree. C.
[0168] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0169] The method of the present invention can provide the compound
of Formula XI-b in any suitable yield. For example, the compound of
Formula XI-b can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0170] The method of the present invention can provide the compound
of Formula XI-b in any suitable purity. For example, the compound
of Formula XI-b can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula XI-b can be prepared in at least about 95%
purity. In some embodiments, the compound of Formula XI-b can be
prepared in at least about 98% purity. In some embodiments, the
compound of Formula XI-b can be prepared in at least about 99%
purity.
[0171] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-b:
##STR00063##
wherein the method comprises forming a reaction mixture having
BCL.sub.3, Et.sub.2N, MeOH, Celite.RTM., and the compound of
Formula XI-a
##STR00064##
under conditions suitable to prepare the compound of Formula XI-b.
In certain embodiments, the method of preparing Formula XI-b is
performed between about -30.degree. C. and about 0.degree. C. In
another embodiment, the method of preparing Formula XI is performed
between about -20.degree. C. and about 0.degree. C.
[0172] In some embodiments, the present invention provides a method
of preparing a compound of Formula XI-c:
##STR00065##
wherein the method comprises forming a reaction mixture having a
solvent, a reagent, an acid, and the compound of Formula XI-b
##STR00066##
under conditions suitable to prepare the compound of Formula
XI-c.
[0173] Any suitable solvent can be used in the method of making the
compound of Formula XI-c. For example, the solvent can be acetone,
MeOH, DCM, THF, MeTHF, Et.sub.2O, MeCN, EtCN, toluene, benzene,
chlorobenzene, nitrobenzene, flurorbenzene, methanol, ethanol,
2-propanol, propanol, butanol, MTBE, EtOAc, iPrOAc, Me2O, (TMS)2O,
acetone, 2-butanone, chloroform, 1,2-dichloroethane, diglyme,
dioxane, acetic acid, formic acid, trifluoroacetic acid,
methylisobutylketone, DMAc, DMF, NMP, DMSO; or combinations
thereof. In some embodiments, the solvent can be acetone.
[0174] The solvent can be present in any suitable amount. For
example, the solvent can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula XI-b, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
solvent can also be present in an amount of from about 0.1 to about
10.0 eq. (mol/mol) to the compound of Formula XI-b, such as of from
about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0 to about
2.0 eq. (mol/mol). In some embodiments, the solvent can be present
in an amount from about 0.1 to 1.0 eq. (mol/mol) to the compound of
Formula XI-b. In some embodiments, the solvent can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula XI-b.
[0175] Any suitable reagent can be used in the method of making the
compound of Formula XI-c. For example, the reagent can be
2,2-dimethoxypropane, acetone, 2-methoxypropene,
2,2-diethylpropane, 2-ethoxypropene, 2,2-dimethyl-1,3-dioxolane,
2,2-dimethyl-1,3-dioxane; or combinations thereof. In some
embodiments, the reagent can be 2,2-dimethoxypropane.
[0176] The reagent can be present in any suitable amount. For
example, the reagent can be present in an amount of at least 0.1
eq. (mol/mol) to the compound of Formula XI-b, such as about 0.1,
0.5, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The
reagent can also be present in an amount of from about 0.1 to about
10.0 eq. (mol/mol) to the compound of Formula XI-b, such as of from
about 0.1 to about 3.0 eq. (mol/mol), or of from about 1.0 to about
2.0 eq. (mol/mol). In some embodiments, the reagent can be present
in an amount from about 0.1 to 1.0 eq. (mol/mol) to the compound of
Formula XI-b. In some embodiments, the reagent can be present in an
amount of from about 1.0 to about 2.0 eq. (mol/mol) to the compound
of Formula XI-b.
[0177] Any suitable acid can be used in the method of making the
compound of Formula XI-c. For example, the acid can be TMSOTf,
TMSOTf, TBSOTf, TESOTf, BF.sub.3, BF.sub.3-OEt.sub.2, BCl.sub.3,
BF.sub.3-THF, MgCl.sub.2, MgI.sub.2, MgBr.sub.2,
MgBr.sub.2-OEt.sub.2, ZnCl.sub.2, ZnBr.sub.2, ZnI.sub.2, LiCl,
LiBr, LiI, AlCl.sub.3, AlBr.sub.3, AlI.sub.3,
Me.sub.2Si(OTf).sub.2, Et.sub.2Si(OTf).sub.2,
Pr.sub.2Si(OTf).sub.2, iPr.sub.2Si(OTf).sub.2,
(tBu).sub.2Si(OTf).sub.2, (C.sub.6F.sub.5).sub.3B, MeSiCl.sub.3,
Me.sub.2SiCl.sub.2, SiCl.sub.4, TMSCl, TMSI, TMSVr, TBSCl, TBSBr,
TBSI, TESCl, TESBr, TESI, SmCl.sub.3, SmBr.sub.3, SmI.sub.2,
SmI.sub.3, ScI.sub.3, ScBr.sub.3, ScI.sub.3, Sm(OTf).sub.3,
Sc(OTf).sub.3, TiCl.sub.4, Ti(OiPr).sub.4, Ti(OiPr).sub.3Cl,
Ti(OiPr).sub.2Cl.sub.2, Ti(OiPr)Cl.sub.3, Zn(BF.sub.4).sub.2,
LiBF.sub.4, Mg(BF.sub.4).sub.2, ZrCl.sub.4, FeCl.sub.2, FeCl.sub.3,
FeBr.sub.2, FeBr.sub.3, FeI.sub.2, FeI.sub.3, Cu(OTf),
Cu(OTf).sub.2, 4-toluenesulfonylchoride, benzenesulfonylchlopride,
4-toluenesulfonyl triflate, benzenesulfonyl triflate,
methylsulfonyl chloride, methylsulfonic anhydrate, InCl.sub.3,
InBr.sub.3, InI.sub.3, In(OTf).sub.3, Mg(SO.sub.4).sub.2,
NaSO.sub.4, dicyclohexylcarbodiimide,
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, benzenesulfonic
acid, HCl, 4-toluenesulfonic acid, triflic acid, trifluoroacetic
acid, 4-nitrobenzolic acid, methylsoulfonic acid, sulfuric acid,
phosphoric acid, HBr, acetic acid, formic acid, HI, TFA,
benzenesulfonic acid, HCl, 4-toluenesulfonic acid, triflic acid,
trifluoroacetic acid, 4-nitrobenzoic acid, methylsoulfonic acid,
sulfuric acid, phosphoric acid, HBr, acetic acid, formic acid, HI,
trifluoromethylsulfonic acid, 4-fluorobenzoic acid, pivalic acid,
HBF.sub.4, nitric acid, 4-chloro-benzoic acid, pentafluorophenol,
HPF.sub.6, Camphorsulfonic acid; or combinations thereof. In some
embodiments, the acid can be sulfuric acid.
[0178] The acid can be present in any suitable amount. For example,
the acid can be present in an amount of at least 0.0 eq. (mol/mol)
to the compound of Formula XI-b, such as about 0.0, 0.5, 1.0, 2, 3,
4, 5, 6, 7, 8, 9, or about 10.0 eq. (mol/mol). The acid can also be
present in an amount of from about 0.0 to about 10.0 eq. (mol/mol)
to the compound of Formula XI-b, such as of from about 0.0 to about
3.0 eq. (mol/mol), or of from about 1.0 to about 2.0 eq. (mol/mol).
In some embodiments, the acid can be present in an amount from
about 0.0 to 1.0 eq. (mol/mol) to the compound of Formula XI-b. In
some embodiments, the acid can be present in an amount of from
about 1.0 to about 2.0 eq. (mol/mol) to the compound of Formula
XI-b.
[0179] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -50.degree. C. to about 50.degree. C., or of
from about 0.degree. C. to about 50.degree. C., or of from about
0.degree. C. to about 40.degree. C., or of from about 0.degree. C.
to about 30.degree. C., or of from about 0.degree. C. to about
25.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about 0.degree. C. to about 23.degree. C. In
some embodiments, the temperature of the reaction mixture can be of
from about 0.degree. C. to about 25.degree. C. In some embodiments,
the temperature of the reaction mixture can be of from about
0.degree. C. to about 30.degree. C.
[0180] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0181] The method of the present invention can provide the compound
of Formula XI-c in any suitable yield. For example, the compound of
Formula XI-c can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0182] The method of the present invention can provide the compound
of Formula XI-c in any suitable purity. For example, the compound
of Formula XI-c can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula XI-b can be prepared in at least about 95%
purity. In some embodiments, the compound of Formula XI-b can be
prepared in at least about 98% purity. In some embodiments, the
compound of Formula XI-b can be prepared in at least about 99%
purity.
[0183] In some embodiments, the method comprises preparing the
compound of Formula XI-c:
##STR00067##
wherein the method comprises forming a reaction mixture having
acetone, 2,2-dimethoxypropane, sulfuric acid, and the compound of
Formula XI-b
##STR00068##
under conditions suitable to prepare the compound of Formula XI-c.
In certain embodiments, the method of preparing Formula XI-c is
performed between about 0.degree. C. and about 30.degree. C. In
another embodiment, the method of preparing Formula XI is performed
between about 10.degree. C. and about 30 C.
[0184] C. Addition of Prodrug Moiety
[0185] The present invention also provides a method of coupling a
prodrug moiety to a nucleoside to provide a compound of the present
invention. In some embodiments, the present invention provides a
method of preparing a compound of Formula VIII:
##STR00069##
wherein the method includes forming a reaction mixture including a
coupling agent, a non-nucleophilic base, a compound of Formula
IX:
##STR00070##
and a compound of Formula X:
##STR00071##
under conditions suitable to form the compound of Formula VIII,
wherein each R.sup.a is H or PG, each PG group is a hydroxy
protecting group, or both PG groups are combined to form
--C(R.sup.19).sub.2--, R.sup.e1 and R.sup.e2 are each independently
H, C.sub.1-C.sub.6 alkyl or benzyl, R.sup.f is H, C.sub.1-C.sub.8
alkyl, benzyl, C.sub.3-C.sub.6 cycloalkyl, or
--CH.sub.2--C.sub.3-C.sub.6 cycloalkyl, R.sup.19 is H,
C.sub.1-C.sub.8 alkyl, phenyl or substituted phenyl, and LG is a
leaving group.
[0186] Any suitable coupling agent can be used in the method of
making the compound of Formula VIII, as described above for the
method of making the compound of Formula V. In some embodiments,
the coupling agent can be a magnesium coupling agent. In some
embodiments, the coupling agent can be MgCl.sub.2, iPrMgCl,
tBuMgCl, PhMgCl, or combinations thereof. In some embodiments, the
coupling agent can be MgCl.sub.2.
[0187] Any suitable non-nucleophilic base can be used in the method
of making the compound of Formula VIII. Representative
non-nucleophilic bases include, but are not limited to,
triethylamine, diisopropylethyl amine, N,N-diethylaniline,
pyridine, 2,6-lutidine, 2,4,6-collidine, 4-dimethylaminopyridine,
and quinuclidine. In some embodiments, the non-nucleophilic base
can be di-isopropyl ethyl amine (DIPEA).
[0188] The protecting groups PG can be any suitable hydroxy
protecting groups, as described above for the method of making the
compound of Formula V. Exemplary protecting groups PG can be
benzyl, or the PG groups can be combined to form an acetonide.
Exemplary acetonides include, but are not limited to acetonide and
benzylidene acetal. In some embodiments, the hydroxy protecting
groups of hydroxy groups on adjacent carbons can be combined to
form acetonide. In some embodiments, the PG groups are combined to
form --C(R.sup.19).sub.2--. In some embodiments, each R.sup.a is
the protecting group PG where the PG groups are combined to form
--C(Me).sub.2-.
[0189] When the R.sup.e group is C.sub.1-C.sub.8 alkyl, each
R.sup.e can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, each R.sup.e group
can be methyl.
[0190] When the R.sup.f group is C.sub.1-C.sub.8 alkyl, R.sup.f can
be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-buty,
t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl, neohexyl,
septyl or octyl. In some embodiments, the R.sup.f group can be
methyl, ethyl, isopropyl, t-butyl, or iso-hexyl. When the R.sup.f
group is C.sub.3-C.sub.6 cycloalkyl, R.sup.f can be cyclopropyl,
cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R.sup.f
can be cyclobutyl, cyclopentyl or cyclohexyl.
[0191] When the R.sup.19 group is C.sub.1-C.sub.8 alkyl, R.sup.19
can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, the R.sup.19 group
can be methyl.
[0192] The leaving group can be any suitable leaving group.
Suitable leaving groups LG include, but are not limited to,
chloride, bromide, mesylate, tosylate, triflate,
4-nitrobenzenesulfonate, 4-chlorobenzenesulfonate, 4-nitrophenoxy,
pentafluorophenoxy, etc. In some embodiments, the leaving group LG
can be 4-nitrophenoxy or pentafluorophenoxy. In some embodiments,
the leaving group LG can be 4-nitrophenoxy.
[0193] In some embodiments, each R.sup.a is PG where the PG groups
are combined to form --C(R.sup.19).sub.2--, R.sup.f is
C.sub.1-C.sub.8 alkyl, R.sup.19 is C.sub.1-C.sub.8 alkyl, and the
leaving group LG is 4-nitrophenoxy or pentafluorophenoxy.
[0194] In some embodiments, the coupling agent is MgCl.sub.2, and
the non-nucleophilic base is di-isopropyl ethyl amine.
[0195] In some embodiments, the compound of Formula VIII can be
##STR00072##
In some embodiments, the compound of Formula VIII can be
##STR00073##
In some embodiments, the compound of Formula VIII can be
##STR00074##
[0196] In some embodiments, the method of making the compound
Formula VIII includes forming the reaction mixture including
MgCl.sub.2, DIPEA, the compound of Formula IX:
##STR00075##
and the compound of Formula X:
##STR00076##
under conditions suitable to form the compound of Formula VIII:
##STR00077##
[0197] When the R.sup.a groups of the compound of Formula VIII are
the hydroxy protecting groups PG, the method can include the
additional step of removing the protecting groups to form the
compound of Formula VIII where each R.sup.a is H. In some
embodiments, the method of preparing the compound of Formula VIII
includes forming a second reaction mixture including a deprotection
agent and the compound Formula VIII wherein each R.sup.a group is
the protecting group PG, under suitable conditions to form the
compound of Formula VIII where each R.sup.a is H. The deprotection
agent can be any suitable agent to remove the protecting groups PG
such as hydrogen and a hydrogenation catalyst, or acid. For
example, if the protecting group PG is benzyl, the deprotection
agent can be hydrogen and platinum on carbon. Alternatively, when
the protecting group PG is an acetonide, the deprotection agent can
be an acid. Representative acids include, but are not limited to,
acetic acid, glacial acetic acid, trifluoroacetic acid (TFA),
hydrochloric acid, concentrated hydrochloric acid, and others. In
some embodiments, the method of preparing the compound of Formula
VIII includes forming a second reaction mixture including an acid
and the compound Formula VIII wherein the R.sup.a groups are
combined to form --C(R.sup.19).sub.2--, under suitable conditions
to form the compound of Formula VIII where each R.sup.a is H. In
some embodiments, the acid can be hydrochloric acid.
[0198] Any suitable solvent can be used in the method of the
present invention. Representative solvents include, but are not
limited to, pentane, pentanes, hexane, hexanes, heptane, heptanes,
petroleum ether, cyclopentanes, cyclohexanes, benzene, toluene,
xylene, trifluoromethylbenzene, halobenzenes such as chlorobenzene,
fluorobenzene, dichlorobenzene and difluorobenzene, methylene
chloride, chloroform, acetone, ethyl acetate, diethyl ether,
tetrahydrofuran, acetonitrile, or combinations thereof. In some
embodiments, the solvent can be acetonitrile.
[0199] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -78.degree. C. to about 100.degree. C., or of
from about -50.degree. C. to about 100.degree. C., or of from about
-25.degree. C. to about 50.degree. C., or of from about -10.degree.
C. to about 25.degree. C., or of from about 0.degree. C. to about
20.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about 0.degree. C. to about 20.degree.
C.
[0200] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0201] The method of the present invention can provide the compound
of Formula VIII in any suitable yield. For example, the compound of
Formula VIII can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0202] The method of the present invention can provide the compound
of Formula VIII in any suitable purity. For example, the compound
of Formula VIII can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula VIII can be prepared in at least 95% purity. In
some embodiments, the compound of Formula VIII can be prepared in
at least 98% purity. In some embodiments, the compound of Formula
VIII can be prepared in at least 99% purity.
[0203] In some embodiments, the present invention provides the
compound
##STR00078##
[0204] In some embodiments, the present invention provides a method
of preparing a compound of Formula VIII:
##STR00079##
wherein the method comprises forming a reaction mixture including a
coupling agent, a non-nucleophilic base, a compound of Formula
IX-a:
##STR00080##
and a compound of Formula X:
##STR00081##
under conditions suitable to form the compound of Formula VIII,
wherein R.sup.a is independently H or a hydroxy protecting group,
or two R.sup.a on adjacent carbons can be combined to form a
--C(R.sup.19).sub.2-- group, R.sup.35 is independently H or a
hydroxy protecting group, or two R.sup.35 on adjacent carbons can
be combined to form a --C(R.sup.19).sub.2-- group, R.sup.19 is H or
C.sub.1-C.sub.8 alkyl, R.sup.e1 and R.sup.e2 are each independently
H, C.sub.1-C.sub.6 alkyl or benzyl, R.sup.f is H, C.sub.1-C.sub.8
alkyl, benzyl, C.sub.3-C.sub.6 cycloalkyl, or
--CH.sub.2--C.sub.3-C.sub.6 cycloalkyl, R.sup.19 is H,
C.sub.1-C.sub.8 alkyl, phenyl or substituted phenyl, and LG is a
leaving group.
[0205] Any suitable coupling agent can be used in the method of
making the compound of Formula VIII, as described above for the
method of making the compound of Formula V. In some embodiments,
the coupling agent can be a magnesium coupling agent. In some
embodiments, the coupling agent can be MgCl.sub.2, iPrMgCl,
tBuMgCl, PhMgCl, or combinations thereof. In some embodiments, the
coupling agent can be MgCl.sub.2.
[0206] Any suitable non-nucleophilic base can be used in the method
of making the compound of Formula VIII. Representative
non-nucleophilic bases include, but are not limited to,
triethylamine, diisopropylethyl amine, N,N-diethylaniline,
pyridine, 2,6-lutidine, 2,4,6-collidine, 4-dimethylaminopyridine,
and quinuclidine. In some embodiments, the non-nucleophilic base
can be di-isopropyl ethyl amine (DIPEA).
[0207] The hydroxy protecting groups, as described above for the
method of making the compound of Formula V. Exemplary hydroxy
protecting group can be benzyl, SiR.sub.3, wherein each R group can
be hydrogen, alkyl, alkenyl, cycloalkyl, phenyl, or other silicon
containing groups, or the PG groups can be combined to form an
acetonide. Exemplary silanes include, but are not limited to
tert-butyldimethylsilyl (TBS). Exemplary acetonides include, but
are not limited to acetonide and benzylidene acetal. In some
embodiments, the hydroxy protecting groups of hydroxy groups on
adjacent carbons can be combined to form acetonide. In some
embodiments, the PG groups are combined to form
--C(R.sup.19).sub.2--. In some embodiments, each R.sup.a is the
protecting group PG where the PG groups are combined to form
--C(Me).sub.2-. In other embodiments, PG is a SiR.sub.3. In other
embodiments, PG is tert-butyldimethylsilyl (TBS).
[0208] When the R.sup.e group is C.sub.1-C.sub.8 alkyl, each
R.sup.e can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, each R.sup.e group
can be methyl.
[0209] When the R.sup.f group is C.sub.1-C.sub.8 alkyl, R.sup.f can
be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-buty,
t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl, neohexyl,
septyl or octyl. In some embodiments, the R.sup.f group can be
methyl, ethyl, isopropyl, t-butyl, or iso-hexyl. When the R.sup.f
group is C.sub.3-C.sub.6 cycloalkyl, R.sup.f can be cyclopropyl,
cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R.sup.f
can be cyclobutyl, cyclopentyl or cyclohexyl.
[0210] When the R.sup.19 group is C.sub.1-C.sub.8 alkyl, R.sup.19
can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl,
sec-buty, t-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, isohexyl,
neohexyl, septyl or octyl. In some embodiments, the R.sup.19 group
can be methyl.
[0211] When the R.sup.35 group is a hydroxy protecting group,
R.sup.35 can be any example protecting group described in
Protective Groups in Organic Chemistry, Peter G. M. Wuts and
Theodora W. Greene, 4th Ed., 2006. In some embodiments, the
R.sup.35 group can be benzyl. In some embodiments, the R.sup.35
group can be TBS.
[0212] The leaving group can be any suitable leaving group.
Suitable leaving groups LG include, but are not limited to,
chloride, bromide, mesylate, tosylate, triflate,
4-nitrobenzenesulfonate, 4-chlorobenzenesulfonate, 4-nitrophenoxy,
pentafluorophenoxy, etc. In some embodiments, the leaving group LG
can be 4-nitrophenoxy or pentafluorophenoxy. In some embodiments,
the leaving group LG can be 4-nitrophenoxy.
[0213] In some embodiments, each R.sup.a is PG where the PG groups
are combined to form --C(R.sup.19).sub.2--, R.sup.f is
C.sub.1-C.sub.8 alkyl, R.sup.19 is C.sub.1-C.sub.8 alkyl, and the
leaving group LG is 4-nitrophenoxy or pentafluorophenoxy.
[0214] In some embodiments, the coupling agent is MgCl.sub.2, and
the non-nucleophilic base is di-isopropyl ethyl amine.
[0215] In some embodiments, the compound of Formula VIII can be
##STR00082##
In some embodiments, the compound of Formula VIII can be
##STR00083##
In some embodiments, the compound of Formula VIII can be
##STR00084##
[0216] In some embodiments, the method of making the compound
Formula VIII comprises forming the reaction mixture including
MgCl.sub.2, DIPEA, the compound of Formula IX:
##STR00085##
and the compound of Formula X:
##STR00086##
under conditions suitable to form the compound of Formula VIII:
##STR00087##
[0217] When the R.sup.a groups of the compound of Formula VIII are
the hydroxy protecting groups PG, the method can include the
additional step of removing the protecting groups to form the
compound of Formula VIII where each R.sup.a is H. In some
embodiments, the method of preparing the compound of Formula VIII
comprises forming a second reaction mixture including a
deprotection agent and the compound Formula VIII wherein each
R.sup.a group is the protecting group PG, under suitable conditions
to form the compound of Formula VIII where each R.sup.a is H. The
deprotection agent can be any suitable agent to remove the
protecting groups PG such as hydrogen and a hydrogenation catalyst,
or acid. For example, if the protecting group PG is benzyl, the
deprotection agent can be hydrogen and platinum on carbon.
Alternatively, when the protecting group PG is an acetonide, the
deprotection agent can be an acid. Representative acids include,
but are not limited to, acetic acid, glacial acetic acid,
trifluoroacetic acid (TFA), hydrochloric acid, concentrated
hydrochloric acid, formic acids, toluenesulfonic acid, sulfuric
acid, and others. Additional representative acids include, but are
not limited to those found in Greene, T. W.; Wuts, P. G. M.
Protective Groups In Organic Synthesis, 4th Ed., John Wiley &
Sons: New York, 2006. In some embodiments, the method of preparing
the compound of Formula VIII comprises forming a second reaction
mixture including an acid and the compound Formula VIII wherein the
R.sup.a groups are combined to form --C(R.sup.19).sub.2--, under
suitable conditions to form the compound of Formula VIII where each
R.sup.a is H. In some embodiments, the acid can be hydrochloric
acid. Alternatively, when the protecting group PG is SiR.sub.3, the
deprotection agent can be TBAF, pyridine HF, HCl, TsOH, camphor
sulfonic acid, AcCl in MeOH, BF.sup.3 OEt.sup.2, TFA, AGOG, Formic
Acid, HBr, F, HF, Et.sub.3N--HF, KF--H.sub.2O, KHF.sub.2, NaF, LiF,
LiCl, LiBr, LiI, and others.
[0218] Any suitable solvent can be used in the method of the
present invention.
[0219] Representative solvents include, but are not limited to,
pentane, pentanes, hexane, hexanes, heptane, heptanes, petroleum
ether, cyclopentanes, cyclohexanes, benzene, toluene, xylene,
trifluoromethylbenzene, halobenzenes such as chlorobenzene,
fluorobenzene, dichlorobenzene and difluorobenzene, methylene
chloride, chloroform, acetone, ethyl acetate, diethyl ether,
tetrahydrofuran, acetonitrile, or combinations thereof. In some
embodiments, the solvent can be acetonitrile. In some embodiments,
the solvent can be MeCN. In some embodiments, the solvent can be
tetrahydrofuran.
[0220] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -78.degree. C. to about 100.degree. C., or of
from about -50.degree. C. to about 100.degree. C., or of from about
-25.degree. C. to about 50.degree. C., or of from about -10.degree.
C. to about 25.degree. C., or of from about 0.degree. C. to about
20.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about 0.degree. C. to about 20.degree.
C.
[0221] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can also be exposed to any suitable
environment, such as atmospheric gasses, or inert gasses such as
nitrogen or argon.
[0222] The method of the present invention can provide the compound
of Formula VIII in any suitable yield. For example, the compound of
Formula VIII can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0223] The method of the present invention can provide the compound
of Formula VIII in any suitable purity. For example, the compound
of Formula VIII can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula VIII can be prepared in at least about 95%
purity. In some embodiments, the compound of Formula VIII can be
prepared in at least about 98% purity. In some embodiments, the
compound of Formula VIII can be prepared in at least about 99%
purity.
In some embodiments, the compound of Formula VIII can be
##STR00088##
[0224] In some embodiments, the method of making the compound
Formula VIII comprises forming the reaction mixture including
MgCl.sub.2, DIPEA, the compound of Formula IX-a.sup.2:
##STR00089##
and the compound of Formula X:
##STR00090##
under conditions suitable to form the compound of Formula VIII:
##STR00091##
[0225] The method can include the additional step of removing the
protecting groups to form the compound of Formula VIII where each
TBS is H.
[0226] The reaction mixture of the method can be at any suitable
temperature. For example, the temperature of the reaction mixture
can be of from about -78.degree. C. to about 100.degree. C., or of
from about -50.degree. C. to about 100.degree. C., or of from about
-25.degree. C. to about 50.degree. C., or of from about -10.degree.
C. to about 25.degree. C., or of from about 0.degree. C. to about
20.degree. C. In some embodiments, the temperature of the reaction
mixture can be of from about 0.degree. C. to about 20.degree.
C.
[0227] The reaction mixture of the method can be at any suitable
pressure. For example, the reaction mixture can be at atmospheric
pressure. The reaction mixture can be also be exposed to any
suitable environment, such as atmospheric gasses, or inert gasses
such as nitrogen or argon.
[0228] The method of the present invention can provide the compound
of Formula VIII in any suitable yield. For example, the compound of
Formula VIII can be prepared in a yield of at least about 50%, 55,
60, 65, 70, 75, 80, 85, 90 or at least about 95%.
[0229] The method of the present invention can provide the compound
of Formula VIII in any suitable purity. For example, the compound
of Formula VIII can be prepared in a purity of at least about 90,
95, 96, 97, 98 or at least about 99%. In some embodiments, the
compound of Formula VIII can be prepared in at least about 95%
purity. In some embodiments, the compound of Formula VIII can be
prepared in at least about 98% purity. In some embodiments, the
compound of Formula VIII can be prepared in at least about 99%
purity.
[0230] D. Preparation of Formula X-b by Crystallization-Induced
Dynamic Resolution
[0231] In one embodiment, there is provided a method for the
crystallization-induced dynamic resolution of (2S)-2-ethylbutyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate (Formula
X-a):
##STR00092##
to provide (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate
(Formula X-b). The method comprises subjecting a solution
comprising: a) a suitable solvent; b) a suitable base; c)
(2S)-2-ethylbutyl 2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)
propanoate; and, optionally, d) one or more seed crystals of
(S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate, to
conditions that provide for the epimerization of the phosphorus
center, under conditions that also provide selective
crystallization of (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino) propanoate.
[0232] The crystallization can be carried out in any suitable
solvent. For example, it can be carried out in an aprotic organic
solvent, or in a mixture thereof. For example, the aprotic organic
solvent may comprise ethyl acetate, methyl acetate, propyl acetate,
isopropyl acetate, diethyl ether, diisopropyl ether,
tetrahydrofuran, dichloromethane, acetone, methyl ethyl ketone,
methyl tert-butylether, toluene, or acetonitrile, or a mixture
thereof. In one embodiment, the solvent comprises acetonitrile.
[0233] The resolution can be carried out in the presence of any
suitable base. For example, the resolution can be carried out in
the presence of a base selected from
1,5-diazobicyclo[4.3.0]non-5-ene (DBN),
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU),
7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (MTBD), triethylamine
(Et.sub.3N), Hunig's Base (iPr.sub.2NEt), tetramethylguanidine, a
Verkade base (e.g.,
2,8,9-triisopropyl-2,5,8,9-tetraaza-1-phosphabicyclo[3.3.3]undecane,
and 2,8,9-triisobutyl-2,5,8,9-tetraaza-1-phosphabicyclo
[3.3.3]undecane), a metal carbonate (e.g., M.sub.xCO.sub.3), a
metal phenoxide (M.sup.+-OPh), and PhOTMS in combination with a
fluoride ion source (e.g., R.sub.4N.sup.+-F, TASF
(tris(dimethylamino) sulfonium difluorotrimethylsilicate), or TBAT
(tetrabutylammonium triphenyldifluorosilicate), and mixtures
thereof, wherein each M is a suitable metal such as an alkali metal
or an alkaline earth metal, and each R is, for example, a
(C.sub.1-C.sub.6) alkyl. In one specific embodiment, the base is
DBU.
[0234] The resolution can also be carried out at any suitable
temperature, for example, a temperature in the range of from about
0.degree. C. to about 50.degree. C. In one specific embodiment, the
resolution is carried out at a temperature of about 0.degree.
C.
[0235] In one specific embodiment, the resolution is carried out in
the presence of phenol.
[0236] The percentage of (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl) amino) propanoate in
the starting diastereomeric mixture can be anywhere in the range
from about 0% to about 99%. In one embodiment of the invention, the
percentage of (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate in the
starting diastereomeric mixture is in the range from about 0% to
about 20%. In one embodiment, the percentage of Compound
(S)-2-ethylbutyl 2-(((S)-(4-nitrophenoxy)(phenoxy)
phosphoryl)amino)propanoate in the starting diastereomeric mixture
is in the range from about 20% to about 99%. In one embodiment, the
percentage of Compound (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate in the
starting diastereomeric mixture is in the range from about 50% to
about 99%. In one embodiment, the final Compound (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate is at
least about 90%, about 95%, about 97%, or about 99%
diastereomerically pure. In one embodiment, the final Compound
(S)-2-ethylbutyl 2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino)
propanoate contains less than 1% of any diastereomeric impurities.
In one embodiment, the final Compound (S)-2-ethylbutyl
2-(((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)amino) propanoate is
free of any detectable diastereomeric impurities.
Examples
[0237] Certain abbreviations and acronyms are used in describing
the experimental details. Although most of these would be
understood by one skilled in the art, Table 1 contains a list of
many of these abbreviations and acronyms.
TABLE-US-00001 TABLE 1 List of abbreviations and acronyms.
Abbreviation Meaning Ac.sub.2O acetic anhydride AIBN 2,2'-azobis
(2-methylpropionitrile) Bn benzyl BnBr benzylbromide BSA bis
(trimethylsilyl) acetamide BzCl benzoyl chloride CDI carbonyl
diimidazole DABCO 1,4-diazabicyclo[2.2.2]octane DBN
1,5-diazabicyclo[4.3.0]non-5-ene DDQ
2,3-dichloro-5,6-dicyano-1,4-benzoquinone DBU
1,5-diazabicyclo[5.4.0]undec-5-ene DCA dichloroacetamide DCC
dicyclohexylcarbodiimide DCM dichloromethane DMAP
4-dimethylaminopyridine DME 1,2-dimethoxyethane DMTCl
dimethoxytrityl chloride DMSO dimethylsulfoxide DMTr 4,
4'-dimethoxytrityl DMF dimethylformamide EtOAc ethyl acetate ESI
electrospray ionization EtOAc ethyl acetate HMDS
hexamethyldisilazane HPLC High pressure liquid chromatography LDA
lithium diisopropylamide LRMS low resolution mass spectrum MCPBA
meta-chloroperbenzoic acid MeCN acetonitrile MeOH methanol MMTC
mono methoxytrityl chloride m/z or m/e mass to charge ratio
MH.sup.+ mass plus 1 MH.sup.- mass minus 1 MsOH methanesulfonic
acid MS or ms mass spectrum MTBE tert-butylmethyl ether NBS
N-bromosuccinimide Ph phenyl rt or r.t. room temperature TBAF
tetrabutylammonium fluoride THF tetrahydrofuran TMSCl
chlorotrimethylsilane TMSBr bromotrimethylsilane TMSI
iodotrimethylsilane TMSOTf (trimethylsily) trifluoromethylsulfonate
TEA triethylamine TBA tributylamine TBAP tributylammonium
pyrophosphate TBSCl t-butyldimethylsilyl chloride TEAB
triethylammonium bicarbonate TFA trifluoroacetic acid TLC or tlc
thin layer chromatography Tr triphenylmethyl Tol 4-methylbenzoyl
Turbo Grignard 1:1 mixture of isopropylmagnesium chloride and
lithium chloride .delta. parts per million down field from
tetramethylsilane
[0238] E. Preparation of Compounds
Example 1. (2S)-ethyl 2-(chloro(phenoxy)phosphorylamino)propanoate
(Chloridate A)
##STR00093##
[0240] Ethyl alanine ester hydrochloride salt (1.69 g, 11 mmol) was
dissolved in anhydrous CH.sub.2Cl.sub.2 (10 mL) and the mixture
stirred with cooling to 0.degree. C. under N.sub.2(g). Phenyl
dichlorophosphate (1.49 mL, 10 mmol) was added followed by dropwise
addition of Et.sub.3N over about 10 min. The reaction mixture was
then slowly warmed to RT and stirred for about 12 h. Anhydrous
Et.sub.2O (50 mL) was added and the mixture stirred for about 30
min. The solid that formed was removed by filtration, and the
filtrate concentrated under reduced pressure. The residue was
subjected to silica gel chromatography eluting with 0-50% EtOAc in
hexanes to provide intermediate A. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.39-7.27 (m, 5H), 4.27 (m, 3H), 1.52 (m, 3H),
1.32 (m, 3H). .sup.31P NMR (121.4 MHz, CDCl.sub.3) .delta. 8.2,
7.8.
Example 2. (2S)-2-ethylbutyl
2-(chloro(phenoxy)phosphorylamino)propanoate (Chloridate B)
##STR00094##
[0242] The 2-ethylbutyl alanine chlorophosphoramidate ester B was
prepared using the same procedure as chloridate A except
substituting 2-ethylbutyl alanine ester for ethyl alanine ester.
The material is used crude in the next reaction. Treatment with
methanol or ethanol forms the displaced product with the requisite
LCMS signal.
Example 3. (2S)-isopropyl
2-(chloro(phenoxy)phosphorylamino)propanoate (Chloridate
##STR00095##
[0244] The isopropyl alanine chlorophosphoramidate ester C was
prepared using the same procedure as chloridate A except
substituting isopropyl alanine ester for the ethyl alanine ester.
The material is used crude in the next reaction. Treatment with
methanol or ethanol forms the displaced product with the requisite
LCMS signal.
Example 4. (2R, 3R, 4S,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxy-
methyl)tetrahydrofuran-2-carbonitrile (Compound 1)
##STR00096##
[0246] The preparation of (2R, 3R, 4S,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxy-
methyl)tetrahydrofuran-2-carbonitrile is described below.
##STR00097##
[0247] The commercially available lactol (10 g, 23.8 mmol) was
dissolved in anhydrous DMSO (30 mL) under N.sub.2(g). Ac.sub.2O (20
mL) was added and the resultant reaction mixture stirred at RT for
about 48 h. The reaction mixture was poured onto ice H.sub.2O (500
mL) and the mixture stirred for 20 min. The mixture was extracted
with EtOAc (3.times.200 mL) and the combined organic extracts were
then washed with H.sub.2O (3.times.200 mL). The organic extract was
dried over anhydrous MgSO.sub.4, filtered and concentrated under
reduced pressure. The residue was dissolved in CH.sub.2Cl.sub.2 and
subjected to silica gel chromatography eluting with 25% EtOAc in
hexanes to provide the lactone. .sup.1H NMR (400 MHz, DMSO) .delta.
7.30-7.34 (m, 13H), 7.19-7.21 (m, 2H), 4.55-4.72 (m, 6H), 4.47 (s,
2H), 4.28 (d, J=3.9 Hz, 1H), 3.66 (m, 2H). LCMS m/z 436.1
[M+H.sub.2O], 435.2 [M+OH]- Tr=2.82 min. HPLC Tr=4.59 [2-98% ACN in
H2) over 5 min at 2 mL/min flow.
##STR00098##
[0248] The bromopyrazole (prepared according to WO2009/132135) (0.5
g, 2.4 mmol) was suspended in anhydrous THF (10 mL) under
N.sub.2(g). The suspension was stirred and TMSCl (0.67 mL, 5.28
mmol) was added. The mixture was stirred for 20 min. at RT and then
cooled to about -78.degree. C. after which time a solution of
n-BuLi (6 mL, 1.6 N in hexanes, 9.6 mmol) was added slowly. The
reaction mixture was stirred for 10 min. at about -78.degree. C.
and then the lactone (1 g, 2.4 mmol) was added via syringe. When
the reaction was complete as measured by LCMS, AcOH was added to
quench the reaction. The mixture was concentrated under reduced
pressure and the residue dissolved in a mixture of CH.sub.2Cl.sub.2
and H.sub.2O (100 mL, 1:1). The organic layer was separated and
washed with H.sub.2O (50 mL). The organic layer was then dried over
anhydrous MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was subjected to silica gel chromatography
eluting with 0-50% EtOAc in hexanes to provide the product as a 1:1
mixture of anomers. LCMS m/z 553 [M+H].
##STR00099##
[0249] The hydroxy nucleoside (1.1 g, 2.0 mmol) was dissolved in
anhydrous CH.sub.2Cl.sub.2 (40 mL) and the solution cooled with
stirring to about -78.degree. C. under N.sub.2(g). TMSCN (0.931 mL,
7 mmol) was added and the mixture stirred for a further 10 min.
TMSOTf (1.63 mL, 9.0 mmol) was slowly added to the reaction and the
mixture stirred for 1 h. The reaction mixture was then diluted with
CH.sub.2Cl.sub.2 (120 mL) and aqueous NaHCO.sub.3 (120 mL) was
added to quench the reaction. The reaction mixture was stirred for
a further 10 min and the organic layer separated. The aqueous layer
was extracted with CH.sub.2Cl.sub.2 (150 mL) and the combined
organic extracts dried over anhydrous MgSO.sub.4, filtered and
concentrated under reduced pressure. The residue was dissolved in a
minimal amount of CH.sub.2Cl.sub.2 and subjected to silica gel
chromatography eluting with a gradient of 0-75% EtOAc and hexanes
to provide the tribenzyl cyano nucleoside as a mixture of anomers.
.sup.1H NMR (300 MHz, CD.sub.3CN) .delta. 7.94 (s, 0.5H), 7.88 (s,
0.5H), 7.29-7.43 (m, 13H), 7.11-7.19 (m, 1H), 6.82-6.88 (m, 1H),
6.70-6.76 (m, 1H), 6.41 (bs, 2H), 5.10 (d, J=3.9 Hz, 0.5H), 4.96
(d, J=5.1 Hz, 0.5H), 4.31-4.85 (m, 7H), 4.09-4.18 (m, 2H),
3.61-3.90 (m, 2H). LCMS m/z 562 [M+H].
##STR00100##
[0250] The tribenzyl cyano nucleoside (70 mg, 0.124 mmol) was
dissolved in anhydrous CH.sub.2Cl.sub.2 (2 mL) and cooled to about
-20.degree. C. under N.sub.2(g). A solution of BCl.sub.3 (1N in
CH.sub.2Cl.sub.2, 0.506 mL, 0.506 mmol) was added and the reaction
mixture stirred for 1 h. at -78.degree. C. When the reaction was
complete by LC/MS, MeOH was added to quench the reaction. The
reaction mixture was allowed to warm to RT and the solvent removed
under reduced pressure. The residue was subjected to C18 reverse
phase HPLC, eluting for 5 min with H.sub.2O (0.1% TFA), followed by
a gradient of 0-70% MeCN in H.sub.2O (0.1% TFA) over 35 min, to
elute the .alpha.-anomer, and .beta.-anomer 1. (.alpha.-anomer)
.sup.1H NMR (300 MHz, D.sub.2O) .delta. 7.96 (s, 1H), 7.20 (d,
J=4.8 Hz, 1H), 6.91 (d, J=4.8 Hz, 1H), 4.97 (d, J=4.4 Hz, 1H),
4.56-4.62 (m, 1H), 4.08-4.14 (m, 1H), 3.90 (dd, J=12.9, 2.4 Hz,
1H), 3.70 (dd, J=13.2, 4.5 Hz, 1H). (.beta.-anomer) .sup.1H NMR
(400 MHz, DMSO) .delta. 7.91 (s, 1H), 7.80-8.00 (br s, 2H),
6.85-6.89 (m, 2H), 6.07 (d, J=6.0 Hz, 1H), 5.17 (br s, 1H), 4.90
(br s, 1H), 4.63 (t, J=3.9 Hz, 1H), 4.02-4.06 (m, 1H), 3.94 (br s,
1H), 3.48-3.64 (m, 2H). LCMS m/z 292.2 [M+H], 290.0 [M-H]. Tr=0.35
min. 13C NMR (400 MHZ, DMSO), 156.0, 148.3, 124.3, 117.8, 117.0,
111.2, 101.3, 85.8, 79.0, 74.7, 70.5, 61.4. HPLC Tr=1.32 min
Example 4-a. (2R, 3R, 4S,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxy-
methyl)tetrahydrofuran-2-carbonitrile (Compound 1)
##STR00101##
[0252] The preparation of (2R, 3R, 4S,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxy-
methyl)tetrahydrofuran-2-carbonitrile is described below.
Preparation of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol Using
LaCl.sub.3-2LiCl
##STR00102##
[0254] A solution of 7-iodopyrrolo[2,1-f][1,2,4]triazin-4-amine
(7.5 g, 28.8 mmol, 1.0 equiv) was prepared in THF (67 mL). The
solution was cooled to about 0.degree. C., and TMSCl (3.3 mL, 30.3
mmol, 1.05 equiv) was added. The reaction mixture was stirred for
about 30 min, and then PhMgCl (2 M in THF; 28 mL, 56.8 mmol, 1.97
equiv) was added while maintaining an internal temperature below
5.degree. C. The reaction mixture was agitated at about 0.degree.
C. for about 35 min, and then cooled to about -15.degree. C.
iPrMgCl (2 M in THF, 14 mL, 30.2 mmol, 1.05 equiv) was then added
while maintaining an internal temperature below about -10.degree.
C. After approximately 15 minutes at about -15.degree. C.,
LaCl.sub.3-2LiCl (0.6 M in THF, 50 mL, 14.4 mmol, 0.5 equiv) was
added while maintaining an internal temperature below about
-15.degree. C. The reaction mixture was agitated for about 25 min
at about -20.degree. C.
[0255] In a separate flask, a solution of
(3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)dihydrofuran-2(3H)-one
(10.0 g, 23.9 mmol, 0.83 equiv) was prepared in THF (45 mL). The
solution was cooled to about -20.degree. C., and then transferred
to the Grignard solution while maintaining an internal temperature
below about -15.degree. C. The resulting reaction mixture was
agitated at about -20.degree. C. for about 30 min.
[0256] The reaction was quenched with 2 M HCl (53 mL), and the
mixture warmed to about 15.degree. C. iPrOAc (38 mL) was added, and
the organic and aqueous phases were separated. The bottom aqueous
layer was discharged, and the upper organic layer was washed
sequentially with 2.5 wt % NaHCO.sub.3 (53 mL), 2.5 wt %
NaHCO.sub.3 (53 mL), and 10 wt % NaCl (53 mL).
[0257] The organic phase was concentrated to about 45 mL, and then
diluted with iPrOAc (75 mL). The solution was concentrated again to
about 45 mL, and then diluted with iPrOAc (23 mL). The solution was
concentrated to about 45 mL, and then filtered over a pad of
Celite. The filtered solution was concentrated to about 26 mL, and
then diluted with MTBE (75 mL). After 2 h, heptane (23 mL) was
slowly added and the slurry was stirred at about 25.degree. C. for
about 2 h, and was then cooled to about -5.degree. C. over about 8
h. The solids were isolated by filtration, and the filter cake was
washed with MTBE/heptane (4:1, 23 mL). The solids were dried in a
vacuum oven at no more than about 35.degree. C. to afford
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol.
Preparation of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol Using CeCl.sub.3
##STR00103##
[0259] The iodopyrazole (5.02 g, 19.3 mmol) was dissolved in THF
(45 g) and the solution was cooled to about 0.degree. C. with
stirring. TMSCl (2.04 g, 18.7 mmol) was added, and after about 1 h
phenyl magnesium chloride (2.0 M in THF, 19.9 g, 38.2 mmol) was
added. The reaction mixture was cooled to about -20.degree. C. and
iso-propyl magnesium chloride (2.0 M in THF, 9.99 g, 20.5 mmol) was
added slowly. After about 30 min, the reaction mixture was
transferred to a mixture of anhydrous cerium chloride (4.75 g, 19.3
mmol) in THF (22 g) at about -20.degree. C. After about 1.5 h a
solution of lactone (6.73 g, 16.1 mmol) in THF (22 g) was added
slowly, and the resulting reaction mixture was stirred for about 1
h. 2 M HCl (41 g) was added, the mixture was warmed to about
15.degree. C., and iso-propyl acetate (35 g) was added. The layers
were separated and the organic layer was washed with 2.5%
NaHCO.sub.3 (2.times.40 g), 10% NaCl (1.times.35 g) and
concentrated to about 30 mL volume. iso-Propyl acetate (44 g) was
charged and the solution was concentrated to about 30 mL volume.
iso-Propyl acetate (43 g) was charged and the solution was
concentrated to about 30 mL volume. The solution was filtered and
the filtrate was concentrated to about 18 mL volume.
tert-Butylmethyl ether (37 g) was added followed by product seed
crystals (10.7 mg). After about 14 h n-heptane (10.5 g) was added
and the mixture was cooled to about -5.degree. C. and filtered. The
solids were washed with tert-butylmethyl ether (9 g) at about
-5.degree. C. and dried under vacuum at about 34.degree. C. for
about 15 h to provide the product.
Preparation of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol Using CeCl.sub.3 and
iPrMgCl--LiCl
##STR00104##
[0261] The iodopyrazole (5.03 g, 19.3 mmol) was dissolved in THF
(45 g) and the solution was cooled to about 0.degree. C. with
stirring under N.sub.2(g). TMSCl (2.06 g, 19.0 mmol) was added, and
after about 1 h phenyl magnesium chloride (2.0 M in THF, 20.23 g,
38.8 mmol) was added. The reaction mixture was cooled to about
-20.degree. C. and iso-propyl magnesium chloride-lithium chloride
complex (2.0 M in THF, 15.37 g, 21.0 mmol) was added slowly. After
about 1 h, the reaction mixture was transferred to a mixture of
cerium chloride (4.77 g, 19.4 mmol) in THF (22 g) at about
-20.degree. C. After about 1 h a solution of lactone (6.75 g, 16.1
mmol) in THF (23 g) was added slowly, and the resulting reaction
mixture was stirred for about 1.5 h. 2 M HCl (40 g) was added, the
mixture was warmed to about 15.degree. C. and iso-propyl acetate
(35 g) was added. The layers were separated and the organic layer
was washed with 2.5% NaHCO.sub.3 (2.times.40 g), 10% NaCl
(1.times.36 g) and concentrated to about 30 mL volume. iso-Propyl
acetate (44 g) was added and the solution was concentrated to about
30 mL volume. The solution was filtered and the filtrate was
concentrated to about 18 mL volume. tert-Butylmethyl ether (37 g)
was added followed by product seed crystals (10.5 mg). After about
14 h n-heptane (11 g) was added and the mixture was cooled to about
-5.degree. C. and filtered. The solids were washed with
tert-butylmethyl ether (9 g) at about -5.degree. C. and dried under
vacuum at about 34.degree. C. for about 15 h to provide the
product.
Preparation of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol Using YCl.sub.3
##STR00105##
[0263] The iodopyrazole (4.99 g, 19.2 mmol) was dissolved in THF
(44 g) and the solution was cooled to about 0.degree. C. with
stirring. TMSCl (2.45 mL, 19.4 mmol) was added, and after about 30
min phenyl magnesium chloride (2.0 M in THF, 20.29 g, 39.0 mmol)
was added. The reaction mixture was cooled to about -20.degree. C.
and iso-propyl magnesium chloride (2.0 M in THF, 9.85 g, 20.1 mmol)
was added slowly. After about 30 min, the reaction mixture was
transferred into a mixture of anhydrous yttrium chloride (3.76 g,
19.3 mmol) and lactone (6.68 g, 16.0 mml) in THF (24 g) at about
-20.degree. C. After about 2.5 h 2 M HCl (30 g) was added, the
mixture was warmed to about 15.degree. C., and iso-propyl acetate
(22 g) was added. The layers were separated and the organic layer
was washed with 2.5% NaHCO.sub.3 (2.times.40 g), 10% NaCl
(1.times.35 g) and concentrated to about 30 mL volume. iso-Propyl
acetate (44 g) was charged and the solution was concentrated to
about 30 mL volume. iso-Propyl acetate (45 g) was charged and the
solution was concentrated to about 30 mL volume. The solution was
filtered and the filtrate was concentrated to about 18 mL volume.
tert-Butylmethyl ether (37 g) was added followed by product seed
crystals (11.5 mg). After about 1 h n-heptane (15 mL) was added and
the mixture was cooled to about -5.degree. C. and agitated for
about 17 h. The slurry was filtered and the solids were washed with
a tert-butylmethyl ether (8 g)/n-heptane (2 g) mixture precooled to
about -5.degree. C. The resulting solids were dried under vacuum at
about 34.degree. C. for about 22 h to afford the product.
Preparation of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol Using NdCl.sub.3
##STR00106##
[0265] The iodopyrazole (5.02 g, 19.3 mmol) was dissolved in THF
(38 g) and the solution was cooled to about 0.degree. C. with
stirring under N.sub.2(g). TMSCl (2.45 mL, 19.4 mmol) was added,
and after about 1 h phenylmagnesium chloride (2.0 M in THF, 19.75
g, 38.0 mmol) was added. The reaction mixture was cooled to about
-20.degree. C. and iso-propylmagnesium chloride (2.0 M in THF, 9.40
g, 19.2 mmol) was added slowly. After about 1.5 h, the reaction
mixture was transferred into a mixture of anhydrous neodymium (III)
chloride (4.03 g, 16.1 mmol) and lactone (6.70 g, 16.0 mml) in THF
(22 g) at about -20.degree. C. After about 1.5 h the reaction
mixture was warmed to -10.degree. C. and, after an additional 2 h,
2 M HCl (36 g) was added. The mixture was warmed to about
15.degree. C. and iso-propyl acetate (23 g) was added. The layers
were separated and the organic layer was washed with 2.5%
NaHCO.sub.3 (2.times.44 g), 10% NaCl (1.times.41 g) and
concentrated to about 30 mL volume. iso-Propyl acetate (44 g) was
charged and the solution was concentrated to about 30 mL volume.
iso-Propyl acetate (45 g) was charged and the solution was
concentrated to about 30 mL volume. The solution was filtered and
the filtrate was concentrated to about 18 mL volume.
tert-Butylmethyl ether (37 g) was added followed by product seed
crystals (11.9 mg). After about 1 h n-heptane (15 mL) was added and
the mixture was cooled to about -5.degree. C. and agitated for
about 15 h. The slurry was filtered and the solids were washed with
a tert-butylmethyl ether (8 g)/n-heptane (11 g) mixture precooled
to about -5.degree. C. The resulting solids were dried under vacuum
at about 34.degree. C. for about 25 h to afford the product.
Preparation of
(2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyl-
oxy)-5-((benzyloxy)methyl)tetrahydrofuran-2-carbonitrile
##STR00107##
[0267] To a pre-cooled (-40.degree. C.) solution of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol (10.0 grams, 18.1
mmols, 1.0 equiv.) in DCM (100 mL) was charged trifluoroacetic acid
(6.19 grams, 54.3 mmols, 3.0 equiv.), followed by a pre-cooled
(-30.degree. C.) solution of TMSOTf (24.1 grams, 108.6 mmols, 6.0
equiv.) and TMSCN (10.8 grams, 108.6 mmols, 6.0 equiv.) in DCM (50
mL) while maintaining the internal temperature below about
-25.degree. C. The reaction mixture was agitated at below about
-30.degree. C. for no less than 10 minutes and quenched into a
pre-cooled (about -10.degree. C.) solution of 20 wt. % KOH aq. (120
mL). The bi-phasic mixture was warmed to ambient temperature. The
organic layer was separated and washed with 10 wt. % NaCl aq.
(3.times.50 mL). The organic phase was filtered, concentrated under
vacuum to about 50 mL, re-diluted with toluene (200 mL) and
concentrated under vacuum to 140 mL at about 50.degree. C. The
solution was seeded with
(2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyl-
oxy)-5-((benzyloxy)methyl)tetrahydrofuran-2-carbonitrile at about
55.degree. C. Agitated at about 55.degree. C. for about an hour and
cooled to about 0.degree. C. over about 6 hours. The solids were
isolated by filtration and the filter cake was washed with toluene
(30 mL). The solids were dried under vacuum at about 50.degree.
C.
Preparation of
(2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyl-
oxy)-5-((benzyloxy)methyl)tetrahydrofuran-2-carbonitrile Via Flow
Chemistry
##STR00108##
[0269] Solutions of
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol (23.0 g in 460.07 g of
DCM), TMSOTf (55.81 g in 138.07 g of DCM) and TMSCN (25.03 g in
138.10 g of DCM) were sequentially pumped, into a tube reactor at
about -40.degree. C. The reaction mixture was collected in a flask,
kept in ice bath, containing 20% KOH aqueous solution (46.91 g KOH
and 210 g of water). The layers were separated and the organic
phase was sequentially washed with 10% KOH aqueous solution (10 g
KOH and 90 mL of water) and with 10% brine (2.times.100 g). The
organic phase was concentrated under vacuum to about 4 volumes,
isopropyl alcohol was charged (162.89 g) and the mixture was
concentrated under vacuum to about 10 volumes. The contents were
warmed to about 60.degree. C., then adjusted to about 0.degree. C.
over about 6.5 h and agitated at about 0.degree. C. for about 15.5
h. The resulting slurry was filtered, the solids were rinsed with
isopropyl alcohol (61.79 g) and then dried at about 50.degree. C.
under reduced pressure overnight to afford the product.
Preparation of (2R, 3R, 4S,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxy-
methyl)tetrahydrofuran-2-carbonitrile
##STR00109##
[0271] The tribenzyl cyano nucleoside (48.8 g, 86.9 mmol, 1.0
equiv.) was dissolved in anhydrous CH.sub.2Cl.sub.2 (244 mL) and
cooled to about -20.degree. C. A solution of BCl.sub.3 (1M in
CH.sub.2Cl.sub.2, 295 mL, 295 mmol, 3.4 equiv.) was added dropwise,
maintaining the internal temperature below about -15.degree. C.
Following addition, the reaction mixture was stirred for 1 h at
about -20.degree. C. MeOH (340 ml) was added dropwise, maintaining
the internal temperature below -15.degree. C. The resulting
solution was distilled to about 250 ml, then refilled with about
250 ml MeOH. The resulting solution was again distilled to about
250 ml, then refilled with about 250 ml MeOH, and finally distilled
to about 125 ml. Water (125 ml) was added, followed by
K.sub.2CO.sub.3 solution (20 wt % in water, 125 ml). The pH was
checked, and found to be .about.3. K.sub.2CO.sub.3 solution was
added (20 wt % in water, 50 ml), and the pH was found to be
.about.8. The resulting slurry was stirred overnight, then filtered
and washed with water (50 ml) and MeOH (50 ml). The wet product
cake was dried overnight at about 40.degree. C. overnight. .sup.1H
NMR (300 MHz, D.sub.2O) .delta. 7.96 (s, 1H), 7.20 (d, J=4.8 Hz,
1H), 6.91 (d, J=4.8 Hz, 1H), 4.97 (d, J=4.4 Hz, 1H), 4.56-4.62 (m,
1H), 4.08-4.14 (m, 1H), 3.90 (dd, J=12.9, 2.4 Hz, 1H), 3.70 (dd,
J=13.2, 4.5 Hz, 1H).
Example 5.
(2R,3R,4R,5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-fl-
uoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile
(Compound 2)
##STR00110##
[0273] The preparation of
(2R,3R,4R,5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-fluoro-4-hyd-
roxy-5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile is described
below.
##STR00111##
2-Deoxy-2-fluoro-4,5-O,O-dibenzyl-D-arabinose
[0274] 1'-Methoxy-2-deoxy-2-fluoro-4,5-O,O-dibenzyl-D-arabinose
(1.0 g, 2.88 mmol) in TFA (13.5 mL) was treated with H.sub.2O (1.5
mL) and the resultant mixture stirred for 5 h. The mixture was then
diluted with EtOAc (100 mL) and treated with saturated NaHCO.sub.3
(50 mL). The organic layer was separated and washed with NaCl (50
mL), dried over anhydrous MgSO.sub.4, filtered and concentrated
under reduced pressure. The residue was subjected to silica gel
chromatography (80 g SiO.sub.2 Combiflash HP Gold Column) eluting
with 0-100% EtOAc in hexanes to afford
2-deoxy-2-fluoro-4,5-O,O-dibenzyl-D-arabinose as a white solid:
R.sub.f=0.52 (25% EtOAc in hexanes). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.30 (m, 10H), 5.35 (m, 1H), 4.68-4.29 (m, 7H),
3.70 (d, J=10.5 Hz, 1H), 3.50 (d, J=10.5 Hz, 2H). .sup.19F NMR
(282.2 MHz, CDCl.sub.3) .delta. -207 (m), -211 (m). LCMS m/z 350
[M+H.sub.2O].
##STR00112##
(3R, 4R,
5R)-4-(benzyloxy)-5-(benzyloxymethyl)-3-fluorodihydrofuran-2(3H)-
-one
[0275] 2-Deoxy-2-fluoro-4,5-O,O-dibenzyl-D-arabinose (4.3 g, 12.8
mmol) was dissolved in CH.sub.2Cl.sub.2 (85 mL) was treated with 4
.ANG. MS (10 g) and pyridinium dichromate (14.4 g, 38.3 mmol). The
resultant mixture was stirred for 24 h and then filtered through a
pad of Celite.RTM.. The eluant was concentrated under reduced
pressure and the residue subjected to silica gel chromatography
(120 g SiO.sub.2 HP Gold Combiflash Column) eluting with 0-100%
EtOAc in hexanes to afford (3R, 4R,
5R)-4-(benzyloxy)-5-(benzyloxymethyl)-3-fluorodihydrofuran-2(3H)-one
as a clear oil (3.5 g, 83%): R.sub.f=0.25 (25% EtOAc in hexanes).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.37 (m, 10H), 5.45 (dd,
J=49, 5.7, Hz, 1H), 4.85 (d, J=11.7 Hz, 1H), 4.52 (m, 4H), 4.29 (d,
J=5.4 Hz, 1H), 2.08 (dd, J=15.3, 10.2 Hz, 2H). .sup.19F NMR (282.2
MHz, CDCl.sub.3) .delta. -216. LCMS m/z 348 [M+H.sub.2O]. HPLC
(6-98% MeCN--H.sub.2O gradient, 0.05% TFA modifier) t.sub.R=5.29
min. Phenomenex Synergi 4 m Hydro-RP 80 A, 50.times.4.60 mm, 4
micron; 2 mL/min flow rate
##STR00113##
(3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-
-(benzyloxymethyl)-3-fluorotetrahydrofuran-2-ol
[0276] 7-Bromopyrrolo[1,2-f][1,2,4]-triazin-4-amine (68 mg, 0.319
mmol) in THF (1.4 mL) was treated with TMSCl (89 .mu.L, 0.703 mmol)
and the mixture stirred for 2 h. The mixture was then cooled to
about -78.degree. C. and treated with nBuLi (1.0 M in hexanes, 1.09
mL, 1.09 mmol). The solution was stirred for about 30 min and then
treated with (3R, 4R,
5R)-4-(benzyloxy)-5-(benzyloxymethyl)-3-fluorodihydrofuran-2(3H)-one
(106 mg, 0.319 mmol) dropwise in THF (1.4 mL). The resultant
mixture was stirred for 30 min and then AcOH (83 .mu.L, 1.44 mmol)
in THF (1.0 mL) was added to quench the reaction. The mixture was
warmed to RT and then concentrated under reduced pressure. The
residue was diluted with EtOAc (100 mL) and washed with saturated
NaCl solution (50 mL). The organic layer was dried over anhydrous
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was subjected to silica gel chromatography (40 g SiO.sub.2
HP Gold Combiflash Column) eluting with 0-100% EtOAc in hexanes
followed by a 0-100% gradient of (20% MeOH in EtOAc) in EtOAc to
afford (3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-(benzylo-
xymethyl)-3-fluorotetrahydrofuran-2-ol as a white solid (68 mg,
44%, 60/40 mixture of .alpha./.beta. isomers). R.sub.f=0.32
(EtOAc). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.05 (s, 1H),
7.86 (s, 1H), 7.81 (s, 1H), 7.64 (s, 1H), 7.26 (m, 10H), 6.95 (m,
1H), 6.71 (m, 1H), 6.08 (m, 1H), 5.34 (m, 1H), 4.65 (m, 6H), 4.71
(m, 2H). .sup.19F NMR (282.2 MHz, CDCl.sub.3) .delta. -211 (m).
LCMS m/z 465 [M+H]. HPLC (6-98% MeCN--H.sub.2O gradient, 0.05% TFA
modifier) t.sub.R=4.37 min. (.alpha.-isomer), 4.54 min.
(.beta.-isomer).
##STR00114##
(3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-
-(benzyloxymethyl)-3-fluorotetrahydrofuran-2-carbonitrile
[0277] (3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-(benzylo-
xymethyl)-3-fluorotetrahydrofuran-2-ol (195 mg, 0.42 mmol) was
dissolved in MeCN (1.4 mL) was treated with TMSCN (336 .mu.L, 2.52
mmol) and In(OTf).sub.3 (708 mg, 1.26 mmol). The solution was
stirred at about 70.degree. C. for 18 h and then cooled to about
0.degree. C. The mixture was treated with saturated NaHCO.sub.3
solution (20 drops) then warmed to RT and diluted with EtOAc (100
mL) and H.sub.2O (50 mL). The organic layer was separated and
washed with saturated NaCl solution (50 mL), dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The residue was
subjected to silica gel chromatography (40 g SiO.sub.2 HP Gold
Combiflash Column) eluting with 0-100% EtOAc in hexanes to afford
(3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-
-(benzyloxymethyl)-3-fluorotetrahydrofuran-2-carbonitrile as a
white solid (60/40 mixture of .alpha./.beta. isomers). Data for
both isomers: R.sub.f=0.53 (EtOAc). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.01 (s, 1H), 7.94 (s, 1H), 7.30 (m, 10H), 7.00
(d, J=4.5 Hz, 1H), 6.93 (d, J=4.8 Hz, 1H), 6.87 (d, J=5.4 Hz, 1H),
6.70 (d, J=4.8 Hz, 1H), 5.85 (dd, J=52, 3.3 Hz, 1H), 5.55 (dd,
J=53, 4.5 Hz, 1H), 4.71 (m, 7H), 3.87 (m, 2H), 3.72 (m, 2H).
.sup.19F NMR (282.2 MHz, CDCl.sub.3) .delta. -196 (m), -203 (m).
LCMS m/z 474 [M+H]. HPLC (6-98% MeCN--H.sub.2O gradient, 0.05% TFA
modifier) t.sub.R=4.98 min.
##STR00115##
(2R, 3R, 4R,
5R)-2-(4-aminopyrrolo[1,2,4][1,2,4]triazin-7-yl)-3-fluoro-4-hydroxy-5-(hy-
droxymethyl)tetrahydrofuran-2-carbonitrile (2)
[0278] (3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-(benzyloxy)-5-(benzylo-
xymethyl)-3-fluorotetrahydrofuran-2-carbonitrile (110 mg, 0.23
mmol) was dissolved in CH.sub.2Cl.sub.2 (1.5 mL) and cooled to
about 0.degree. C. The reaction mixture was treated with BCl.sub.3
(1.0 M in CH.sub.2Cl.sub.2, 766 .mu.L, 0.77 mmol) and stirred for 2
h. The mixture was then cooled to about -78.degree. C. and treated
with Et.sub.3N (340 .mu.L, 2.44 mmol) followed by MeOH (2 mL)
before allowing to warm to RT. The reaction was concentrated under
reduced pressure and then co-evaporated with MeOH (3.times.5 mL).
The residue was then suspended in H.sub.2O (5 mL) and treated with
NaHCO.sub.3 (1 g). The solution was stirred for 10 min and then
concentrated under reduced pressure. The residue was filtered and
washed with MeOH (3.times.10 mL) on a fritted glass funnel (coarse)
and the eluant concentrated under reduced pressure. The residue was
subjected to reverse phase HPLC (6-98% MeCN in H.sub.2O gradient
with 0.05% TFA modifier) to afford (2R, 3R, 4R,
5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-fluoro-4-hydroxy-5-(hy-
droxymethyl)tetrahydrofuran-2-carbonitrile 2 as a white solid and
the .alpha.-isomer. Data for the .beta.-isomer: R.sub.f=0.13 (10%
MeOH in EtOAc). .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.09 (s,
1H), 7.28 (d, J=5.1 Hz, 1H), 7.17 (d, J=5.1 Hz, 1H), 5.42 (dd,
J=53, 3.3 Hz, 1H), 4.20 (m, 2H), 3.99 (d, J=3.6 Hz, 1H), 3.77 (d,
J=3.6 Hz, 1H). .sup.19F NMR (282.2 MHz, CDCl.sub.3) .delta. -197
(m). LCMS m/z 294 [M+H]. HPLC (2-98% MeCN--H.sub.2O gradient, 0.05%
TFA modifier) t.sub.R=1.49 min.
Example 6. (2R, 3R, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-fluoro-2-(hydroxymethy-
l)-5-methyltetrahydrofuran-3-ol (Compound 3)
##STR00116##
[0280] The preparation of (2R, 3R, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-fluoro-2-(hydroxymethy-
l)-5-methyltetrahydrofuran-3-ol is described below.
##STR00117##
[0281] The starting nucleoside (prepared as described in the
synthesis of compound 2) (0.355 g, 0.765 mmol) was dissolved in
anhydrous THF (35 mL) and cooled to about 0.degree. C. with
stirring under N.sub.2(g). A solution of methyl magnesium chloride
(2 mL, 6 mmol) (3N in THF) was added and the resultant mixture
stirred overnight. Acetic acid (7 mmol) was added to quench the
reaction and then the solvents were removed by rotory under reduced
pressure. The residue was re-dissolved in CH.sub.2Cl.sub.2 and the
solution subjected to a plug of silica gel to isolate the product
(0.355 g) as a crude mixture. LC/MS (m/z: 480, M.sup.+1). The crude
material was dissolved in anhydrous CH.sub.2Cl.sub.2 (20 mL) and
placed under N.sub.2(g). The solution was stirred and treated with
methanesulfonic acid (0.2 mL, 2.74 mmol). The reaction mixture was
stirred for about 12 h at RT and then quenched by the addition of
Et.sub.3N (3.5 mmol). The mixture was concentrated under reduced
pressure and the residue subjected to silica gel chromatography to
provide the methyl substituted nucleoside as a 4:1 mixture of beta-
and alpha-anomers respectively. .sup.1H NMR (300 MHz, CD.sub.3CN)
major anomer .delta. 7.87 (s, 1H), 7.27-7.40 (m, 10H), 6.77 (d,
J=4.5 HZ, 1H), 6.70 (d, J=4.5 Hz, 1H), 6.23 (br s, 2H), 5.53 (dd,
J=55, 3.3 Hz, 1H), 4.42-4.75 (m, 4H), 4.19-4.26 (m, 1H), 3.65-4.00
(m, 3H), 1.74 (d, J=3.9 Hz, 3H). .sup.19F NMR (282.2 MHz,
CD.sub.3CN) major anomer .delta. -207 (m, 1F). LCMS m/z 463
[M+H].
##STR00118##
[0282] The benzylated nucleoside material (0.134 g, 0.290 mmol),
Degussa catalyst (0.268 g) and AcOH (30 mL) were mixed together.
The reaction atmosphere was charged with H.sub.2 (g) and the
reaction stirred for about 2 h. The catalyst was removed by
filtration and the mixture concentrated under reduced pressure. The
residue was dissolved in a minimal amount of H.sub.2O and subjected
to reverse phase HPLC (C.sup.18 hydro RP column) to isolate the
.beta.-anomer 3. .sup.1H NMR (300 MHz, D.sub.2O) .delta. 7.87 (s,
1H), 7.22 (d, J=4.8 Hz, 1H), 6.87 (d, J=4.8 Hz, 1H), 5.35 (dd,
J=54, 3.6 Hz, 1H), 3.97-4.10 (m, 2H), 3.81 (dd, J=12.6, 2.1 Hz,
1H), 3.64 (dd, J=12.6, 4.8 Hz, 1H), 1.65 (d, J=4.2 Hz, 3H).
.sup.19F NMR (282.2 MHz, CD.sub.3CN) 8-207 (m, 1F).
[0283] A small amount of alpha anomer was characterized as follows.
.sup.1H NMR (300 MHz, D.sub.2O) .delta. 7.86 (s, 1H), 7.26 (d,
J=4.8 Hz, 1H), 6.85 (d, J=4.8 Hz, 1H), 5.31 (dd, J=54, 3.9 Hz, 1H),
4.39 (ddd, J=26.1, 9.9, 3.6 Hz, 2H), 4.00-4.05 (m, 1H), 3.90 (dd,
J=12.3, 2.1 Hz, 1H), 3.66 (dd, J=12.6, 4.8, 1H), 1.56 (s, 3H).
.sup.19F NMR (282.2 MHz, CD.sub.3CN) 8-198 (dd, J=54, 26 Hz,
1F).
Example 7. (2S)-isopropyl
2-(((((2R,3R,4R,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-4-fluoro-
-3-hydroxy-5-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-
propanoate (Compound 4)
##STR00119##
[0285] The nucleoside 3 (0.011 g, 0.04 mmol) was dissolved in
trimethylphosphate (2 mL) and cooled to 0.degree. C. The mixture
was stirred under an atmosphere of N.sub.2(g) and 1-Methylimidazole
(0.320 mL, 5 mmol) followed by the alaninylmonoisopropyl,
monophenol phosphorchloridate C (0.240 mL, 4.4 mmol) was added. The
reaction mixture was stirred for 2 h. at 0.degree. C. and then
allowed to warm slowly to RT. while monitoring by LC/MS. When
complete by LCMS, the reaction mixture was treated with H.sub.2O (5
mL) and then concentrated under reduced pressure. The residue was
dissolved in CH.sub.2Cl.sub.2 and subjected to silica gel
chromatography eluting with 0-100% EtOAc in hexanes. The product
fractions were collected and concentrated. The residue was
subjected to prep HPLC to yield the alanine isopropyl monoamidate
prodrug 4 as a mixture of isomers. .sup.1H NMR (300 MHz,
CD.sub.3CN) .delta. 7.87 (s, 1H), 7.17-7.44 (m, 5H), 6.71-6.83 (m,
2H), 6.14 (br, s, 2H), 5.38 (dd, J=56, 3.3 Hz, 1H), 4.92-5.01 (m,
1H), 3.86-4.46 (m, 6H), 3.58 (m, 1H), 1.73 (m, 3H), 1.18-1.34 (m,
9H). LCMS m/z 552 [M+H].
Example 8. (2S)-ethyl
2-(((((2R,3R,4R,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-4-fluoro-
-3-hydroxy-5-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-
propanoate (Compound 5)
##STR00120##
[0287] The nucleoside 3 (0.026 g, 0.092 mmol) was dissolved in
trimethylphosphate (2 mL) and cooled to 0.degree. C. The mixture
was stirred under N.sub.2(g) and 1-methylimidazole (0.062 mL, 0.763
mmol) followed by the chloridate A (0.160 g, 0.552 mmol) were
added. The reaction mixture was stirred for 2 h. at 0.degree. C.
and then allowed to warm slowly to RT. H.sub.2O (5 mL) was added to
quench the reaction and then the mixture concentrated under reduced
pressure. The residue was dissolved in CH.sub.2Cl.sub.2 and
subjected to silica gel chromatography eluting with 0-100% EtOAc in
hexanes. The product fractions were collected and concentrated.
Crude product was eluted using 0 to 100 percent EtOAc in hexanes.
The crude product was collected and concentrated under reduced
pressure. The residue was subjected to prep HPLC to yield compound
5. LCMS m/z 538 [M+H].
Example 9. ((2R, 3R, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-4-fluoro-3-hydroxy-5-met-
hyltetrahydrofuran-2-yl)methyl Tetrahydrogen Triphosphate (Compound
6)
##STR00121##
[0289] The nucleoside 3 (0.022 g, 0.056 mmol) was dissolved in
trimethylphosphate (1 mL) and stirred under N.sub.2(g). Phosphorous
oxychloride (0.067 mL, 0.73 mmol) was added and the mixture stirred
for about 2 h. Monitoring by analytical ion-exchange column
determined the time at which >80 percent of monophosphate was
formed. A solution of tributylamine (0.44 mL, 1.85 mmol) and
triethylammonium pyrophosphate (0.327 g, 0.72 mmol) dissolved in
anhydrous DMF (1 mL) was added. The reaction mixture was stirred
for 20 min and then quenched by the addition of 1N triethylammonium
bicarbonate solution in H.sub.2O (5 mL). The mixture was
concentrated under reduced pressure and the residue re-dissolved in
H.sub.2O. The solution was subjected to ion exchange chromatography
to yield the title product compound 6. LCMS m/z 521 [M-H]. Tr=0.41.
HPLC ion exchange TR=9.40 min
Example 10.
(2R,3R,5S)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-hydroxy-5-(hydr-
oxymethyl)-tetrahydrofuran-2-carbonitrile (Compound 7)
##STR00122##
[0291] The preparation of
(2R,3R,5S)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-hydroxy-5-(hydr-
oxymethyl)-tetrahydrofuran-2-carbonitrile is described below.
##STR00123##
[0292]
((3.alpha.R,5S,6.alpha.R)-2,2-dimethyl-tetrahydrofuro[2,3-d][1,3]di-
oxol-5-yl)methanol
[0293] The acetate material (1.2 g, 5.5 mmol) (J. Org. Chem. 1985,
50, 3457, De Bernardo et al) was dissolved in a 1:1 mixture MeOH
and THF (10 mL). A 1N solution of NaOH(aq) (10 mL) was added until
the pH was 13. The reaction mixture was stirred for about 2 h and
then neutralized to pH 8-9 by the addition of AcOH. The mixture was
extracted with EtOAc (10.times.30 mL) and the combined organic
extracts dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was subjected to
silica gel chromatography eluting with 0-70% EtOAc in hexanes to
give the desired product (866 mg, 90%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 5.84 (d, J=3.6 Hz, 1H), 4.78 (t, J=4.5 Hz, 1H),
4.38 (m, 1H), 3.93-3.54 (m, 2H), 2.04-1.84 (m, 2H), 1.52 (s, 3H),
1.33 (s, 3H).
##STR00124##
(3.alpha.R,5S,6.alpha.R)-5-(benzyloxymethyl)-2,2-dimethyl-tetrahydrofuro[-
2,3-d][1,3]dioxole
[0294] Sodium hydride (188 mg, 7.46 mmol) was dissolved in
anhydrous THF (5 mL) and stirred under N.sub.2(g) at RT. The
alcohol (866 mg, 4.97 mmol) was dissolved in anhydrous THF (3 mL)
and then added in portions over 5 min. to the sodium hydride
mixture. The resultant mixture was stirred for about 20 min. and
then benzyl bromide (892 .mu.L, 7.46 mmol) was added. The reaction
was stirred for about 2 h and then poured onto a mixture of ice
cold aqueous NaHCO.sub.3 and EtOAc (30 mL). The organic layer was
separated and then the aqueous layer re-extracted with EtOAc (30
mL). The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was subjected to silica gel chromatography eluting with
0-40% EtOAc in hexanes to give the benzyl ether product. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.35-7.27 (m, 5H), 5.86 (d, J=3.6
Hz, 1H), 4.74 (t, J=4.2 Hz, 1H), 4.60 (s, 2H), 4.42 (m, 1H),
3.69-3.53 (m, 2H), 2.10-2.04 (m, 1H), 1.83-1.77 (m, 1H), 1.52 (s,
3H), 1.33 (s, 3H).
##STR00125##
(3R,5S)-5-(benzyloxymethyl)-tetrahydrofuran-2,3-diol
[0295] The benzyl ether (910 mg, 3.44 mmol) was dissolved in a 1:1
AcOH and H.sub.2O (20 mL) mixture and stirred at about 60.degree.
C. for about 7 h. The mixture was concentrated under reduced
pressure and the residue subjected to silica gel chromatography
eluting with 0-70% EtOAc in hexanes to give the diol product (705
mg, 91%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.36-7.27 (m,
5H), 5.40 (d, J=3.9 Hz, 0.5H), 5.17 (s, 0.5H), 4.67-4.56 (m, 3H),
4.33 (m, 0.5H), 4.24 (d, J=4.8 Hz, 0.5H), 3.71-3.67 (m, 1H),
3.56-3.42 (m, 2H), 2.31-2.22 (m, 1H), 2.08-1.89 (m, 2H).
##STR00126##
(3R,5S)-5-(benzyloxymethyl)-3-hydroxy-dihydrofuran-2(3H)-one
[0296] The diol (705 mg, 3.14 mmol) was dissolved in benzene (30
mL) and treated with a silver carbonate celite mixture (3.46 g,
6.28 mmol). The resultant mixture was stirred at about 80.degree.
C. under N.sub.2(g) for about 2 h. The mixture was then cooled to
RT, filtered and concentrated under reduced pressure. The residue
was subjected to silica gel chromatography eluting with 0-70% EtOAc
in hexanes to give the lactone product. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.39-7.27 (m, 5H), 4.75-4.68 (m, 1H), 4.60-4.49
(m, 2H), 3.74-3.54 (m, 2H), 2.61-2.35 (m, 2H), 2.38-2.28 (m,
1H).
##STR00127##
(3R,
5S)-3-(benzyloxy)-5-(benzyloxymethyl)-dihydrofuran-2(3H)-one
[0297] The lactone (600 mg, 2.7 mmol) was dissolved in EtOAc (30
mL) and treated with silver oxide (626 mg, 2.7 mmol) followed by
benzyl bromide (387 .mu.L, 3.24 mmol). The reaction mixture was
then stirred at about 50.degree. C. under N.sub.2(g) for about 8 h.
Additional silver oxide (300 mg) was then added and the resultant
mixture stirred at about 50.degree. C. for about 16 h. Additional
benzyl bromide (50 uL) and silver oxide (150 mg) were added and the
mixture stirred for an additional about 8 h. The reaction mixture
was allowed to cool, filtered and then concentrated under reduced
pressure. The residue was subjected to silica gel chromatography
eluting with 0-20% EtOAc in hexanes to give the title product.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.39-7.27 (m, 10H), 4.99
(d, J=11.4 Hz, 1H), 4.72 (m, 2H), 4.56 (m, 2H), 4.39 (t, J=8.1 Hz,
1H), 3.72-3.51 (m, 2H), 2.42-2.25 (m, 2H).
##STR00128##
(3R,5S)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-(benzyloxy)-5-(ben-
zyloxymethyl)-tetrahydrofuran-2-ol
[0298] The 7-bromopyrrolo[1,2-f][1,2,4]triazin-4-amine (607 mg,
2.85 mmol) was dissolved in anhydrous THF (10 mL) and stirred under
Ar(g) at RT. TMSCl (1.1 mL, 8.55 mmol) was added dropwise and the
mixture stirred for about 2 h. The reaction was concentrated under
reduced pressure and then dried under high vacuum. The residue was
suspended in THF (20 mL) and stirred under Ar(g) at about
-78.degree. C. A 2.5M n-BuLi solution in hexane (2.28 mL, 5.7 mmol)
was added dropwise over about 10 min. and the resultant mixture
stirred for about 60 min. The lactone (742 mg, 2.37 mmol) dissolved
in anhydrous THF (7 mL) was added to the above mixture over about
20 min. The reaction mixture was stirred for about 2 h. and then
quenched with AcOH until pH was 5-6. The mixture was allowed to
warm to RT and then diluted with EtOAc. The solution was washed
with saturated NaHCO.sub.3 solution, saturated NaCl, dried over
anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure.
The residue was subjected to silica gel chromatography eluting with
0-80% EtOAc in hexanes to give the title product. LCMS m/z 447.2
[M+H], 445.1 [M-H].
##STR00129##
(3R,5S)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3-(benzyloxy)-5-(ben-
zyloxymethyl)-tetrahydrofuran-2-carbonitrile
[0299] The alcohol (250 mg, 0.56 mmol) was dissolved in anhydrous
CH.sub.2Cl.sub.2 (10 mL) and stirred under Ar(g) at about
-15.degree. C. TMSCN (448 .mu.L, 3.36 mmol) was added dropwise and
the mixture stirred for about 10 min. TMSOTf (466 .mu.L, 2.58 mmol)
was added dropwise over 10 min and the resultant mixture stirred
for about 90 min. at about -15.degree. C. Additional TMSCN (224
.mu.L, 3 eq.) and TMSOTf (202 .mu.L, 2 eq.) was added and stirring
continued for about 5 h. Saturated aqueous NaHCO.sub.3 solution was
added to quench the reaction and the mixture stirred for about 10
min. The organic layer was separated and washed with saturated
aqueous NaHCO.sub.3 solution, saturated NaCl solution, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was subjected to silica gel chromatography
eluting with 0-70% EtOAc in hexanes to give the title product. LCMS
m/z 456.3 [M+H], 454.1 [M-H].
##STR00130##
(2R,3R,5S)2-(4-aminopyrrolo[1,2,4][1,2,4]triazin-7-yl)-3-hydroxy-5-(hydro-
xymethyl)-tetrahydrofuran-2-carbonitrile (7)
[0300] The benzyl ether (150 mg, 0.329 mmol) was dissolved in
anhydrous CH.sub.2Cl.sub.2 (2 mL) and the mixture stirred under
Ar(g) at about -20.degree. C. A 1M BCl.sub.3 solution in
CH.sub.2Cl.sub.2 (724 .mu.L, 0.724 mmol) was added dropwise and the
resultant mixture stirred for about 2 h. Additional 1M BCl.sub.3 in
CH.sub.2Cl.sub.2 (724 .mu.L, 0.724 mmol) was added and stirring
continued for 2 h. The mixture was then cooled to about -78.degree.
C. and slowly treated with a 2:1 mixture of Et.sub.3N and MeOH (3
mL). The mixture was stirred for about 10 min and then treated with
MeOH (10 mL). The reaction was allowed to warm to RT and then
concentrated under reduced pressure. The residue was dissolved in
MeOH and concentrated under reduced pressure. The residue was
dissolved in MeOH again and treated with solid NaHCO.sub.3. The
mixture was stirred for about 5 min and then the solid removed by
filtration. The solution was concentrated under reduced pressure
and subjected to preparative HPLC to provide the desired product 7.
.sup.1H NMR (300 MHz, D.sub.2O) .delta. 7.71 (s, 1H), 6.75 (d,
J=4.5 Hz, 1H), 6.65 (d, J=4.8 Hz, 1H), 4.91 (t, J=6.3 Hz, 1H), 4.57
(m, 1H), 3.67-3.47 (m, 2H), 2.18 (m, 2H). LCMS m/z 276.1 [M+H],
274.0 [M-H].
Example 11. (2S)-isopropyl
2-((((2R,3S,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-3-
,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)-phosphorylamino)propano-
ate (Compound 8)
##STR00131##
[0302] The nucleoside 1 (45 mg, 0.15 mmol) was dissolved in
anhydrous trimethyl phosphate (0.5 mL) and the solution stirred
under N.sub.2(g) at about 0.degree. C. Methyl imidazole (36 .mu.L,
0.45 mmol) was added to the solution. Chlorophosphoramidate C (69
mg, 0.225 mmol) was dissolved in anhydrous THF (0.25 mL) and added
dropwise to the nucleoside mixture. When the reaction was complete
by LCMS, the reaction mixture was diluted with EtOAc and washed
with saturated aqueous NaHCO.sub.3 solution, saturated NaCl, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was subjected to silica gel
chromatography eluting with 0-5% MeOH in CH.sub.2Cl.sub.2 followed
by preparative HPLC to give the product. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.95 (m, 1H), 7.31-6.97 (m, 7H), 4.94 (m, 1H),
4.78 (m, 1H), 4.43 (m, 3H), 4.20 (m, 1H), 3.80 (d, 1H), 1.30-1.18
(m, 9H). .sup.31P NMR (121.4 MHz, CD.sub.3OD) .delta. 3.8. LCMS m/z
561.0 [M+H], 559.0 [M-H].
Example 12. (2S)-2-ethylbutyl
2-((((2R,3S,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-3-
,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphorylamino)propanoa-
te (Compound 9)
[0303] Compound 9 can be prepared by several methods described
below.
Procedure 1
##STR00132##
[0305] Prepared from Compound 1 and chloridate B according to the
same method as for the preparation of compound 8. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 7.87 (m, 1H), 7.31-7.16 (m, 5H), 6.92-6.89
(m, 2H), 4.78 (m, 1H), 4.50-3.80 (m, 7H), 1.45-1.24 (m, 8H),
0.95-0.84 (m, 6H). .sup.31P NMR (121.4 MHz, CD.sub.3OD) .delta.
3.7. LCMS m/z 603.1 [M+H], 601.0 [M-H].
Procedure 2
##STR00133##
[0306] (2S)-2-ethylbutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
Propanoate
[0307] (2S)-2-ethylbutyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate (1.08 g,
2.4 mmol) was dissolved in anhydrous DMF (9 mL) and stirred under a
nitrogen atmosphere at RT.
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile (350 mg, 1.2 mmol)
was added to the reaction mixture in one portion. A solution of
t-butylmagnesium chloride in THF (1M, 1.8 mL, 1.8 mmol) was then
added to the reaction dropwise over about 10 minutes. The reaction
was stirred for about 2 h, at which point the reaction mixture was
diluted with ethyl acetate (50 mL) and washed with saturated
aqueous sodium bicarbonate solution (3.times.15 mL) followed by
saturated aqueous sodium chloride solution (15 mL). The organic
layer was dried over anhydrous sodium sulfate and concentrated
under reduced pressure. The resulting oil was purified with silica
gel column chromatography (0-10% MeOH in DCM) to afford
(2S)-2-ethylbutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
propanoate (311 mg, 43%, 1:0.4 diastereomeric mixture at
phosphorus) as a white solid. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.85 (m, 1H), 7.34-7.23 (m, 2H), 7.21-7.09 (m, 3H),
6.94-6.84 (m, 2H), 4.78 (d, J=5.4 Hz, 1H), 4.46-4.33 (m, 2H),
4.33-4.24 (m, 1H), 4.18 (m, 1H), 4.05-3.80 (m, 3H), 1.52-1.39 (m,
1H), 1.38-1.20 (m, 7H), 0.85 (m, 6H). .sup.31P NMR (162 MHz,
CD.sub.3OD) .delta. 3.71, 3.65. LCMS m/z 603.1 [M+H], 600.9 [M-H].
HPLC (2-98% MeCN--H.sub.2O gradient with 0.1% TFA modifier over 8.5
min, 1.5 mL/min, Column: Phenomenex Kinetex C18, 2.6 um 100 .ANG.,
4.6.times.100 mm) t.sub.R=5.544 min, 5.601 min
Separation of the (S) and (R) Diastereomers
[0308] (2S)-2-ethylbutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
propanoate was dissolved in acetonitrile. The resulting solution
was loaded onto Lux Cellulose-2 chiral column, equilibrated in
acetonitrile, and eluted with isocratic acetonitrile/methanol (95:5
vol/vol). The first eluting diastereomer had a retention time of
17.4 min, and the second eluting diastereomer had a retention time
of 25.0 min.
[0309] First Eluting Diastereomer is (S)-2-ethylbutyl
2-(((R)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate:
##STR00134##
[0310] .sup.1HNMR (400 MHz, CD.sub.3OD) .delta. 8.05 (s, 1H), 7.36
(d, J=4.8 Hz, 1H), 7.29 (br t, J=7.8 Hz, 2H), 7.19-7.13 (m, 3H),
7.11 (d, J=4.8 Hz, 1H), 4.73 (d, J=5.2 Hz, 1H), 4.48-4.38 (m, 2H),
4.37-4.28 (m, 1H), 4.17 (t, J=5.6 Hz, 1H), 4.08-3.94 (m, 2H),
3.94-3.80 (m, 1H), 1.48 (sep, J=12.0, 6.1 Hz, 1H), 1.34 (p, J=7.3
Hz, 4H), 1.29 (d, J=7.2 Hz, 3H), 0.87 (t, J=7.4 Hz, 6H).
.sup.31PNMR (162 MHz, CD.sub.3OD) .delta. 3.71 (s). HPLC (2-98%
MeCN--H.sub.2O gradient with 0.1% TFA modifier over 8.5 min, 1.5
mL/min, Column: Phenomenex Kinetex C18, 2.6 um 100 .ANG.,
4.6.times.100 mm) t.sub.R=5.585 min.
[0311] Second Eluting Diastereomer is (S)-2-ethylbutyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate:
##STR00135##
[0312] .sup.1HNMR (400 MHz, CD.sub.3OD) .delta. 8.08 (s, 1H),
7.36-7.28 (m, 3H), 7.23-7.14 (m, 3H), 7.08 (d, J=4.8 Hz, 1H), 4.71
(d, J=5.3 Hz, 1H), 4.45-4.34 (m, 2H), 4.32-4.24 (m, 1H), 4.14 (t,
J=5.8 Hz, 1H), 4.08-3.94 (m, 2H), 3.93-3.85 (m, 1H), 1.47 (sep,
J=6.2 Hz, 1H), 1.38-1.26 (m, 7H), 0.87 (t, J=7.5 Hz, 6H).
.sup.31PNMR (162 MHz, CD.sub.3OD) .delta. 3.73 (s). HPLC (2-98%
MeCN--H.sub.2O gradient with 0.1% TFA modifier over 8.5 min, 1.5
mL/min, Column: Phenomenex Kinetex C18, 2.6 um 100 .ANG.,
4.6.times.100 mm) t.sub.R=5.629 min.
Example 13. (2S)-ethyl
2-((((2R,3S,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-3-
,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphorylamino)propanoa-
te (Compound 10)
##STR00136##
[0314] The preparation of (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propan-
oate is described below.
Procedure 1. Preparation Via Chloridate A
##STR00137##
[0316] Prepared from Compound 1 and chloridate A using same method
as for the preparation of compound 8. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.95 (m, 1H), 7.32-6.97 (m, 7H), 4.78 (m, 1H),
4.43-4.08 (m, 6H), 3.83 (m, 1H), 1.31-1.18 (m, 6H). .sup.31P NMR
(121.4 MHz, CD.sub.3OD) .delta. 3.7. LCMS m/z 547.0 [M+H], 545.0
[M-H].
Procedure 2. Preparation Via Nitro-Benzene Compound L
##STR00138##
[0318] Compound 1 (50 mg, 0.17 mmol) was dissolved in NMP-THF (1:1
mL)) and cooled with ice bath. tBuMgCl (0.257 mL, 0.257 mmol) was
then added over about 5 min. The resulting mixture was allowed to
warm to RT and was stirred for about 30 min. Then a solution of
compound L (Prepared according to US20120009147, 74.6 mg, 0.189
mmol) in THF (2 mL) was added. After about 30 min, the reaction
mixture was purified by HPLC (acetonitrile 10 to 80% in water) to
give compound 29 as a yellow solid. The solid was further purified
with silica gel chromatography (MeOH 0 to 20% DCM) to afford
compound 29. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.76 (d,
J=6.0 Hz, 1H), 7.25-7.14 (m, 2H), 7.11-6.99 (m, 3H), 6.87-6.72 (m,
2H), 4.70 (d, J=5.4 Hz, 1H), 4.39-4.24 (m, 2H), 4.20 (dddd, J=9.7,
7.9, 5.1, 2.8 Hz, 1H), 4.10 (dt, J=12.8, 5.5 Hz, 1H), 4.06-3.91 (m,
2H), 3.72 (ddq, J=14.3, 9.3, 7.1 Hz, 1H), 1.17 (dd, J=7.1, 1.0 Hz,
1H), 1.14-1.06 (m, 5H). .sup.31P NMR (162 MHz, CD.sub.3OD) .delta.
3.73, 3.68. MS m/z=547 (M+1).sup.+.
Example 14. (2S)-ethyl
2-((((2R,3R,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-4-
-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphorylamino)pro-
panoate (Compound 11)
##STR00139##
[0320] Compound 11 was prepared from Compound 2 and chloridate A
using same method as for the preparation of compound 8. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.91 (m, 1H), 7.33-7.16 (m, 5H),
6.98-6.90 (m, 2H), 5.59 (m, 1H), 4.50-4.15 (m, 4H), 4.12-3.90 (m,
3H), 1.33-1.18 (m, 6H). .sup.31P NMR (121.4 MHz, CD.sub.3OD)
.delta. 3.8. LCMS m/z 549.0 [M+H], 547.1 [M-H].
Example 15. (2S,2'S)-diethyl
2,2'-((((2R,3S,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyan-
o-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)phosphoryl)bis(azanediyl)dipro-
panoate (Compound 12)
##STR00140##
[0322] The nucleoside 1 (14.6 mg, 0.05 mmol) was dissolved in
anhydrous trimethyl phosphate (0.5 mL) and stirred under N.sub.2(g)
at RT. POCl.sub.3 (9.2 .mu.L, 0.1 mmol) was added and the mixture
stirred for about 60 min. Alanine ethyl ester hydrochloride (61 mg,
0.4 mmol) and then Et.sub.3N (70 .mu.L, 0.5 mmol) was added. The
resultant mixture was stirred for about 15 min. and then additional
Et.sub.3N (70 .mu.l, 0.5 mmol) was added to give a solution pH of
9-10. The mixture was stirred for about 2 h. and then diluted with
EtOAc, washed with saturated aqueous NaHCO.sub.3 solution followed
by saturated aqueous NaCl solution. The organic layer was dried
over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced
pressure. The residue was subjected to preparative HPLC (C18
column) to yield the product 12. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 1H), 7.41 (d, J=4.8 Hz, 1H), 7.18 (d, J=4.8 Hz,
1H), 4.78 (d, J=5.6 Hz, 1H), 4.36 (m, 1H), 4.25-4.08 (m, 7H), 3.83
(m, 2H), 1.33-1.23 (m, 12H). .sup.31P NMR (121.4 MHz, CD.sub.3OD)
.delta. 13.8. LCMS m/z 570.0 [M+H], 568.0 [M-H].
Example 16.
(2S,3R,4S,5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-2-ethynyl-5-(h-
ydroxymethyl)tetrahydrofuran-3,4-diol (Compound 13)
##STR00141##
[0324] The preparation of
(2S,3R,4S,5R)-2-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-2-ethynyl-5-(h-
ydroxymethyl)tetrahydrofuran-3,4-diol is described below.
##STR00142##
[0325] The nucleoside alcohol (0.6 g, 1.08 mmol) (prepared as
described in Compound 1 synthesis) was dissolved in anhydrous THF
(8 mL) and placed under N.sub.2(g). The reaction mixture was
stirred and cooled to about 0.degree. C. and then treated with a
0.5N solution of ethynyl magnesium bromide in THF (17.2 mL, 17.2
mmol). The reaction mixture was stirred overnight at RT. AcOH (1.5
mL) was added to quench the reaction. The mixture was concentrated
under reduced pressure and the residue redissolved in
CH.sub.2Cl.sub.2. The solution subjected to a plug of silica gel
eluting with 0 to 80% EtOAc in Hexanes to provide the title product
as a crude mixture. LCMS m/z 579 [M+H].
##STR00143##
[0326] The crude ethynyl alcohol (0.624 g, 1.08 mmol) was dissolved
in anhydrous CH.sub.2Cl.sub.2 (10 mL) and placed under N.sub.2(g).
The mixture was stirred and sulfonic acid (0.2 mL, 2.74 mmol) was
added. The reaction mixture was stirred for about 12 h. at RT. When
complete by LCMS, Et.sub.3N (0.56 mL) was added to quench the
reaction. The reaction was concentrated under reduced pressure and
the residue subjected to silica gel chromatography eluting with 0
to 75% EtOAc in Hexanes to yield the ethynyl nucleoside as a
mixture of anomers. LCMS m/z 561 [M+H].
##STR00144##
[0327] The tribenzyl nucleoside (0.650 g, 1.16 mmol) was dissolved
in anhydrous CH.sub.2Cl.sub.2 (30 mL) and cooled to -78.degree. C.
under N.sub.2(g). A solution of boron tribromide (1 N in
CH.sub.2Cl.sub.2, 5.5 mL) was added and the reaction mixture
stirred for 1 h. at -78.degree. C. A solution of MeOH (10 mL) and
pyridine (2 mL) was added to quench the reaction and the mixture
was allowed to rise to RT. The mixture was concentrated under
reduced pressure and subjected to preparative HPLC to provide the
.alpha.-anomer (20 mg) and .beta.-anomer 13 (110 mg).
(.beta.-anomer) .sup.1H NMR (300 MHz, DMSO) .delta. 7.81 (s, 1H),
7.76 (br s, 2H), 6.80-6.85 (m, 2H), 5.11 (d, J=7.2 Hz, 1H), 4.90
(d, J=6.0 Hz, 1H), 4.82 (dd, J=7.2, 4.8 Hz, 1H), 4.62 (t, J=6.3 Hz,
1H), 3.95-3.99 (m, 1H), 3.85-3.91 (dd, J=11.4, 5.7 Hz, 1H),
3.61-3.67 (m, 1H), 3.47-3.55 (m, 1H), 3.52 (d, J=0.9 Hz, 1H).
(.alpha.-anomer) .sup.1H NMR (300 MHz, DMSO) .delta. 7.80 (s, 1H),
7.59 (bs, 2H), 6.80 (d, J=4.5 Hz, 1H), 6.54 (d, J=4.2 Hz, 1H), 5.00
(d, J=7.2 Hz, 1H), 4.89 (d, J=4.8 Hz, 1H), 4.74 (t, J=5.7 Hz, 1H),
4.58 (t, J=4.5 Hz, 1H), 4.27 (m, 1H), 3.88 (m, 1H), 3.64-3.72 (m,
1H), 3.51-3.59 (m, 1H), 3.48 (d, J=0.6 Hz, 1H). LCMS m/z 291
[M+H].
Example 17.
(2R,3R,4R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-1,3,4-tris(benzyl-
oxy)hexane-2,5-diol (Compound 14)
##STR00145##
[0329] The preparation of
(2R,3R,4R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-1,3,4-tris(benzyl-
oxy)hexane-2,5-diol is described below.
##STR00146##
[0330] The tribenzyl alcohol from Compound 1 synthesis (0.250 g,
0.453 mmol) was dissolved in anhydrous THF (25 mL) and stirred
under N.sub.2(g). The reaction mixture was cooled to 0.degree. C.
and then a 3.0 N solution of methyl magnesium chloride in THF (1.2
mL, 3.62 mmol) was added. The reaction mixture was stirred
overnight at RT. Acetic acid (1.5 mL) was added to quench the
reaction and then the mixture was concentrated under reduced
pressure. The residue was redissolved in CH.sub.2Cl.sub.2 and
subjected to a plug of silica gel eluting with 0 to 80% EtOAc in
hexanes. The crude product (0.452 g) was then used in the next
reaction without further purification. LCMS m/z 569 [M+H].
##STR00147##
[0331] The crude methyl nucleoside (0.452 g, 0.796 mmol) was
dissolved in anhydrous CH.sub.2Cl.sub.2 (20 mL) and stirred under
N.sub.2(g). Methanesulfonic acid (0.2 mL, 2.78 mmol) was added and
the reaction stirred for about 12 hr at RT. Et.sub.3N (0.56 mL) was
added to quench the reaction and then the mixture concentrated
under reduced pressure. The residue was subjected to silica gel
chromatography eluting with 0 to 75% EtOAc in Hexanes to yield the
product as a mixture of anomers. LCMS m/z 551 [M+H].
##STR00148##
[0332] The tribenzyl nucleoside (0.20 g, 0.364 mmol) was dissolved
in AcOH (30 mL). and charged with Pd/C (Degussa) (400 mg). The
stirred mixture was flushed with N.sub.2(g) three times and then
H.sub.2 (g) was introduced, The reaction was stirred under H.sub.2
(g) for 2 h. and then the catalyst removed by filtration. The
solution was concentrated under reduced pressure and under the
residue was re-dissolved in H.sub.2O. The solution was subjected to
preparative HPLC under neutral conditions to provide the
.alpha.-anomer and .beta.-anomer 14. (.alpha.-anomer) .sup.1H NMR
(300 MHz, D.sub.2O) .delta. 7.81 (s, 1H), 7.22 (d, 1H), 6.75 (d,
1H), 4.47 (d, 1H), 4.25-4.31 (m, 1H), 3.88-4.95 (m, 1H), 3.58-3.86
(dd, 2H), 1.50 (s, 3H). (.beta.-anomer) .sup.1H NMR (300 MHz,
D.sub.2O) .delta. 7.91 (s, 1H), 7.26 (d, 1H), 6.90 (d, 1H), 4.61
(d, 1H), 4.00-4.09 (m, 2H), 3.63-3.82 (dd, 2H), 1.67 (s, 3H). LCMS
m/z 281 [M+H].
Example 18.
S,S'-2,2'-((((2R,3S,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-
-cyano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)phosphoryl)bis(oxy)bis(et-
hane-2,1-diyl) bis(2,2-dimethylpropanethioate) (Compound 15)
##STR00149##
[0334] The nucleoside 1 (0.028 g, 0.096 mmol) was dissolved in
trimethylphosphate (1 mL). The reaction was stirred under
N.sub.2(g) and then treated with 1H-tetrazole (0.021 g, 0.29 mmol).
The reaction mixture was cooled to 0.degree. C. and the phosphane
(Nucleoside Nucleotides, Nucleic acids; 14; 3-5; 1995; 763-766.
Lefebvre, Isabelle; Pompon, Alain; Perigaud, Christian; Girardet,
Jean-Luc; Gosselin, Gilles; et al.) (87 mg, 0.192 mmol) was added.
The reaction was stirred for 2 h. and then quenched with 30%
hydrogen peroxide (0.120 mL). The mixture was stirred for 30 min at
RT and then treated with saturated aqueous sodium thiosulfate (1
mL). The mixture was stirred for 10 min. and then concentrated
under reduced pressure. The residue was subjected to preparative
HPLC to isolate the title product 15. .sup.1H NMR (300 MHz,
CD.sub.3CN) .delta. 7.98 (s, 1H), 6.92 (d, 1H), 6.81 (d, 1H), 6.44
(bs, 2H), 4.82 (m, 2H), 4.47 (m, 1H), 4.24 (m, 2H), 4.00 (m, 4H),
3.80 (bs, 1H), 3.11 (m, 4H), 1.24 (s, 9H). .sup.31P NMR (121.4 MHz,
CD.sub.3CN) .delta. -1.85 (s). LCMS m/z 661 [M+H].
Example 19. S,S'-2,2'-((((2R, 3S, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-ethynyl-3,4-dihydroxyt-
etrahydrofuran-2-yl)methoxy)phosphoryl)bis(oxy)bis(ethane-2,1-diyl)
bis(2,2-dimethylpropanethioate) (Compound 16)
##STR00150##
[0336] Compound 16 was prepared using the same method as compound
15 except substituting compound 13 as the starting nucleoside.
.sup.1H NMR (300 MHz, CD.sub.3CN) .delta. 7.91 (s, 1H), 6.86 (d,
J=4.8 Hz, 1H), 6.76 (d, J=4.5 Hz, 1H), 6.29 (bs, 2H), 4.69 (t,
J=2.7 Hz, 1H), 4.58 (d, J=5.7 Hz, 1H), 4.14-4.33 (m, 5H), 3.99-4.07
(m, 4H), 3.53 (d, J=5.4 Hz, 1H), 3.11 (q, J=5.7 Hz, 4H), 1.22 (s,
18H). LCMS m/z 658.9 [M+]. Tr=2.31
Example 20. ((2R, 3S, 4R,
5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydroxytet-
rahydrofuran-2-yl)methyl Tetrahydrogen Triphosphate (Compound
17)
##STR00151##
[0338] Compound 17 was prepared from compound 1 using a similar
procedure to the preparation of compound 6. The product was
isolated as the sodium salt. .sup.1H NMR (400 MHz, D.sub.2O)
.delta. 7.76 (s, 1H), 6.88 (d, J=4.8 Hz, 1H), 6.73 (d, J=4.4 Hz,
1H), 4.86 (d, J=5.2 Hz, 1H), 4.43 (m, 1H), 4.39 (m, 1H), 4.05 (m,
1H), 3.94 (m, 1H). .sup.31P NMR (121.4 MHz, D.sub.2O) .delta. -5.4
(d, 1P), -10.8 (d, 1P), -21.1 (t, 1P). LCMS m/z 530 [M-H], 531.9
[M+H] Tr=0.22 min. HPLC ion exchange Tr=9.95 min.
Example 21. ((2R, 3S, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-ethynyl-3,4-dihydroxyt-
etrahydrofuran-2-yl)methyl Tetrahydrogen Triphosphate (Compound
18)
##STR00152##
[0340] Compound 18 was prepared from compound 13 using a similar
procedure to the preparation of compound 6. The product was
isolated as the TEA salt. .sup.1H NMR (300 MHz, D.sub.2O) .delta.
7.85 (s, 1H), 7.09 (d, J=4.6 Hz, 1H), 6.95 (d, J=4.7 Hz, 1H), 4.23
(m, 2H), 4.08 (m, 2H), 3.06 (q, J=7.4 Hz, 20H), 1.14 (t, J=7.3 Hz,
30H). .sup.31P NMR (121.4 MHz, D.sub.2O) .delta. -10.8 (d, 1P),
-11.2 (d, 1P), -23.2 (t, 1P). LCMS m/z 530.8 [M+H], Tr=0.46. HPLC
ion exchange Tr=9.40 min.
Example 22. ((2R, 3S, 4R,
5S)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-methylte-
trahydrofuran-2-yl)methyl Tetrahydrogen Triphosphate (Compound
19)
##STR00153##
[0342] Compound 19 was prepared from compound 14 using a similar
procedure to the preparation of compound 6. .sup.1H NMR (400 MHz,
D.sub.2O) .delta. 7.78 (s, 1H), 6.98 (m, 1H), 6.84 (m, 1H), 4.45
(m, 1H), 4.04 (m, 4H), 1.54 (s, 3H). .sup.31P NMR (161 MHz,
D.sub.2O) .delta. -10.6 (m), -23.0 (m). LCMS m/z 521.0 [M+H].
Example 23.
((2R,3R,4R,5R)-5-(4-aminopyrrolo[1,2-f][1,2,4]triazin-7-yl)-5-cyano-4-flu-
oro-3-hydroxytetrahydrofuran-2-yl)methyl Tetrahydrogen Triphosphate
(Compound 20)
##STR00154##
[0344] Compound 20 was prepared from compound 2 using a similar
procedure to the preparation of compound 6. .sup.1H NMR (400 MHz,
D.sub.2O) .delta. 7.78 (s, 1H), 6.93 (d, J=4.4 Hz, 1H), 6.78 (d,
J=4.8 Hz, 1H), 5.45 (dd, J=53, 4.4 Hz, 1H), 4.38-4.50 (m, 2H),
4.13-4.20 (m, 2H). .sup.31P NMR (161 MHz, D.sub.2O) .delta. -5.7
(d, 1P), -11.0 (d, 1P), -21.5 (t, 1P). LCMS m/z 533.9.0 [M+H],
532.0 [M-H] Tr=1.25 min. HPLC ion exchange Tr=11.0 min.
Example 24. (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate (21)
##STR00155##
[0346] The preparation of (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate is described below.
Preparation of (S)-ethyl 2-amino-3-phenylpropanoate
hydrochloride
##STR00156##
[0348] L-Phenylalanine (5 g, 30 mmol) was taken up in EtOH (30 mL).
TMSCl (6.915 mL, 54 mmol) was added to the reaction at RT. The
reaction vessel was fitted with a reflux condenser and the reaction
was placed in an 80.degree. C. bath. The reaction was stirred
overnight. The next day the reaction was cooled to RT, concentrated
under reduced pressure and the resulting residue was taken up in
Et.sub.2O. The resulting slurry was filtered and the isolate solids
were further washed with Et.sub.2O. The washed solids were placed
under high vacuum to yield example (S)-ethyl
2-amino-3-phenylpropanoate hydrochloride. .sup.1N NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.52 (s, 3H), 7.30 (m, 5H), 4.24 (ABX,
J.sub.AX=7.8 Hz, J.sub.BX=6.2 Hz, 1H), 4.11 (m, 2H), 3.17, 3.05
(ABX, J.sub.AB=-14 Hz, J.sub.BX=5.8 Hz, J.sub.AX=7.6 Hz, 2H), 1.09
(t, J=6.8 Hz, 3H).
Preparation of (2S)-ethyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)-3-phenylpropanoate
(Compound D)
##STR00157##
[0350] (S)-ethyl 2-amino-3-phenylpropanoate hydrochloride (1.01 g,
4.41 mmol) was dissolved in DCM (50 mL). This solution was cooled
to about 0.degree. C. and PhOP(O)Cl.sub.2 (0.656 mL, 4.41 mmol) was
added, followed by the slow addition of Et.sub.3N (1.62 mL, 11.5
mmol) over 5 min. The cold bath was removed and the reaction was
allowed to warm to RT and stir over a period of 80 min.
p-NO.sub.2PhOH (0.583 g, 4.19 mmol) was added, followed by more
Et.sub.3N (0.3 mL, 2.1 mmol). The reaction progress was monitored
by LC/MS. Upon completion of the reaction, it was diluted with
Et.sub.2O, and the resulting solids were removed by filtration. The
filtrate was concentrated and compound D was isolated by silica gel
column chromatography (25 g dry load cartridge, 120 g column;
eluent: 100% hexanes ramping to 55% EtOAc in hexanes). .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.17 (m, 2H), 7.33 (m, 2H), 7.09-7.25
(m, 10H), 4.17 (m, 1H), 4.07 (m, 2H), 3.08 (m, 1H), 2.84 (m, 1H),
1.14 (m, 3H). .sup.31P NMR (162 MHz, DMSO-d.sub.6) .delta. -1.479
(s), -1.719 (s). MS m/z=471.01 [M+1].
Preparation of (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate (Compound 21)
##STR00158##
[0352] Compound 1 (0.030 g, 0.103 mmol) was dissolved in DMF (1 mL)
and then THF (0.5 mL) was added. t-BuMgCl (1M/THF, 154.5 .mu.L,
0.154 .mu.mol) was added to the reaction in a drop-wise manner with
vigorous stirring. The resulting white slurry was stirred at RT for
about 30 min. A solution of compound D (0.058 g, 0.124 mmol) in THF
(1 mL) was added in a drop-wise manner to the reaction at RT. The
reaction progress was monitored by LC/MS. When the reaction
progressed to 50% conversion, the reaction was cooled in an ice
bath and quenched with glacial acetic acid (70 .mu.L). The reaction
was concentrated and compound 21 was isolated from the residue by
reverse phase HPLC. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.91 (d, J=4 Hz, 1H), 7.90 (brs, 2H), 7.09-7.30 (m, 8H), 7.01, (t,
J=8.2 Hz, 2H), 6.89 (d, J=4.4 Hz, 1H), 6.82 (t, J=4.4 Hz, 1H), 6.27
(m, 1H), 6.14 (m, 1H), 5.34 (m, 1H), 4.62 (t, J=5.6 Hz, 1H), 4.15
(m, 1H), 3.78-4.01 (m, 6H), 2.92 (m, 1H), 2.78 (m, 1H), 1.04 (m,
3H). .sup.31P NMR (162 MHz, DMSO-d.sub.6) .delta. 3.69 (s), 3.34
(s). MS m/z=623.0 [M+H].
Example 25. (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-0)methoxy)(phenoxy)phosphoryl)amino)-3-meth-
ylbutanoate (22)
##STR00159##
[0354] The preparation of (2S)-ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-met-
hylbutanoate is described below.
Preparation of (2S)-ethyl
3-methyl-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino) butanoate
(Compound E)
##STR00160##
[0356] The (S)-ethyl 2-amino-3-methylbutanoate (0.351 g, 1.932
mmol) was dissolved in DCM (17 mL). This solution was cooled in an
ice bath and PhOP(O)Cl.sub.2 (0.287 mL, 1.932 mmol) was added,
followed by the slow addition of Et.sub.3N (1.62 mL, 11.4 mmol)
over about 5 min. The cold bath was removed and the reaction was
allowed to warm to RT and stir over a period of 1 h. p-NO.sub.2PhOH
(0.255 g, 1.836 mmol) was added, and the reaction progress was
monitored by LC/MS. Upon completion of the reaction, the mixture
was diluted with Et.sub.2O, and the resulting solids were removed
by filtration. The filtrate was concentrated and compound E was
isolated by silica gel column chromatography (12 g dry load
cartridge, 80 g column; eluent: 100% hexanes ramping to 55% EtOAc
in hexanes). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.30 (d,
J=9.2 Hz, 2H), 7.48 (t, J=9.6 Hz, 2H), 7.40 (t, J=7.8 Hz, 2H),
7.20-7.27 (m, 3H), 6.60 (quart, J=11.6 Hz, 1H), 4.01 (m, 2H), 3.61
(m, 1H), 1.93 (m, 1H), 1.11 (m, 3H), 0.79 (m, 6H). .sup.31P NMR
(162 MHz, DMSO-d.sub.6) .delta. -0.342 (s), -0.578 (s). MS
m/z=422.9 [M+H].
Preparation of (2S)-ethyl 2-(((((2R,3
S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydro-
xytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-methylbutanoat-
e (Compound 22)
##STR00161##
[0358] Compound 1 (0.040 g, 0.137 mmol) was dissolved in NMP (1.5
mL) and then THF (0.25 mL) was added. This solution was cooled in
an ice bath and t-BuMgCl (1M/THF, 425.7 .mu.L, 0.426 .mu.mol) was
added in a drop-wise manner with vigorous stirring. The ice bath
was removed and the resulting white slurry was stirred at RT for
about 15 min. A solution of compound E (0.081 g, 0.192 mmol) in THF
(0.5 mL) was added in a drop-wise manner to the reaction at RT. The
reaction progress was monitored by LC/MS. When the reaction
progressed to 50% conversion, the reaction was cooled in an ice
bath and quenched with glacial acetic acid (70 .mu.L). The reaction
was concentrated and compound 22 was semi-purified from the residue
by reverse phase HPLC. The semi-pure material was further purified
by silica gel column chromatography (12 g dry load cartridge, 40 g
column; eluent: 100% EtOAc ramping to 10% MeOH in EtOAc) to yield
compound 22. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.91 (d,
J=1.6 Hz, 1H), 7.88 (brs, 2H), 7.32 (m, 2H), 7.15 (m, 3H), 6.90 (t,
J=4.2 Hz, 1H), 6.84 (d, J=4.8 Hz, 1H), 6.26 (dd, J=13.4, 6.2 Hz,
1H), 5.87 (quart. J=11.2 Hz, 1H), 5.35 (m, 1H), 4.64 (m, 1H), 4.25
(m, 2H), 3.93-4.15 (m, 4H), 3.45 (m, 1H), 1.87 (m, 1H), 1.09-1.16
(m, 3H), 0.70-0.83 (m, 6H). .sup.31P NMR (162 MHz, DMSO-d.sub.6)
.delta. 4.59 (s), 4.47 (s). MS m/z=575.02 [M+H].
Example 26. (S)-isopropyl
2-(((R)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate (23)
##STR00162##
[0360] The preparation of (S)-isopropyl
2-(((R)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate is described below.
##STR00163##
[0361] Compound 1 (60.0 mg, 206 .mu.mop was dissolved in NMP (0.28
mL). THF (0.2 mL) was added followed by tert-butyl magnesium
chloride (1.0M solution in tetrahydrofuran, 0.309 mL) at RT under
an argon atmosphere. After 20 min, a solution of compound F
(Prepared according to Cho, A. et al J. Med. Chem. 2014, 57,
1812-1825., 81 mg, 206 .mu.mop in THF (0.2 mL) was added, and the
resulting mixture was warmed to about 50.degree. C. After 3 h, the
reaction mixture was allowed to cool to RT and was purified
directly by preparatory HPLC (Phenominex Synergi 4u Hydro-RR 80
.ANG. 150.times.30 mm column, 5-100% acetonitrile/water gradient)
to afford compound 23. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.86 (s, 1H), 7.34-7.26 (m, 2H), 7.21-7.12 (m, 3H), 6.91 (d, J=4.6
Hz, 1H), 6.87 (d, J=4.6 Hz, 1H), 4.92 (sept, J=6.3 Hz, 1H), 4.80
(d, J=5.4 Hz, 1H), 4.43-4.34 (m, 1H), 4.33-4.24 (m, 1H), 4.18 (t,
J=5.6 Hz, 1H), 3.82 (dq, J=9.7, 7.1 Hz, 2H), 1.27 (dd, J=7.1, 1.0
Hz, 3H), 1.18 (dd, J=6.3, 4.8 Hz, 6H). .sup.31P NMR (162 MHz,
CD.sub.3OD) .delta. 3.72 (s). LC/MS: t.sub.R=1.39 min, MS
m/z=561.11 [M+H]; LC system: Thermo Accela 1250 UHPLC; MS system:
Thermo LCQ Fleet; Column: Kinetex 2.6u XB-C18 100A, 50.times.4.6
mm; Solvents: ACN with 0.1% acetic acid, water with 0.1% acetic
acid; Gradient: 0 min-2.0 min 2-100% ACN, 2.0 min-3.05 min 100%
ACN, 3.05 min-3.2 min 100%-2% ACN, 3.2 min-3.5 min 2% ACN at 2
.mu.l/min. HPLC: t.sub.R=2.523 min; HPLC system: Agilent 1100
series.; Column: Gemini 5.mu. C18 110A, 50.times.4.6 mm; Solvents:
ACN with 0.1% TFA, Water with 0.1% TFA; Gradient: 0 min-5.0 min
2-98% ACN, 5.0 min-6.0 min 98% ACN at 2 mL/min.
Example 27. (2S)-cyclobutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)
Amino)propanoate (24)
##STR00164##
[0363] The preparation of (2S)-cyclobutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propan-
oate is described below.
Preparation of (2S)-cyclobutyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate (Compound
G)
##STR00165##
[0365] Phenyl dichlorophosphate (1.49 mL, 10 mmol) was dissolved in
10 mL of anhydrous DCM and stirred under atmosphere nitrogen in an
ice bath. L-Alanine isobutyl ester hydrochloride (0.9 g, 5 mmol)
was added in one portion. Triethylamine (765 .mu.L, 5.5 mmol) was
then added dropwise. Reaction stirred for about 1 h. More
Triethylamine (765 .mu.L, 5.5 mmol) was added dropwise and the
reaction was stirred for about 45 min. p-Nitrophenol (1.25 g, 9
mmol) was added in one portion and stirred for about 30 min.
Triethylamine (765 .mu.L, 5.5 mmol) was added and the reaction
mixture was stirred for about 2 h. Additional p-nitrophenol (1.25
g, 9 mmol) and triethylamine (765 .mu.L, 5.5 mmol) were then added,
and the reaction was stirred for another about 2 h. The reaction
mixture was concentrated under reduced pressure. The resulting
crude was diluted with EtOAc and washed twice with 5% aqueous
citric acid solution, followed with saturated aqueous sodium
chloride solution. The organic layer was then dried over anhydrous
sodium sulfate and concentrated under reduced pressure. The crude
residue was purified with silica gel column (0-20-50% EtOAc in
hexanes) to give compound G. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.33-8.23 (m, 2H), 7.52-7.33 (m, 4H), 7.33-7.17 (m, 3H),
4.96-4.85 (m, 1H), 4.07-3.96 (m, 1H), 2.27 (m, 2H), 2.07-1.91 (m,
2H), 1.83-1.70 (m, 1H), 1.70-1.55 (m, 1H), 1.32 (m, 3H). .sup.31P
NMR (162 MHz, CD.sub.3OD) .delta. -1.36, -1.59. MS m/z=420.9
[M+H].
Preparation (2S)-cyclobutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propan-
oate (Compound 24)
##STR00166##
[0367] Compound 1 (58 mg, 0.2 mmol) was mixed with compound G (101
mg, 0.24 mmol) in 2 mL of anhydrous DMF. Magnesium chloride (42 mg,
0.44 mmol) was added in one portion. The reaction mixture was
heated to about 50.degree. C. DIPEA (87 .mu.L, 0.5 mmol) was added,
and the reaction was stirred for about 2 h at about 50.degree. C.
The reaction mixture was cooled to room temperature, was diluted
with EtOAc and was washed with 5% aqueous citric acid solution
followed by saturated aqueous sodium chloride solution. The organic
layer was then dried over anhydrous sodium sulfate and concentrated
under reduced pressure. The crude residue was purified with silica
gel column (0-2-5% MeOH in DCM) to afford compound 24. .sup.1H NMR
(400 MHz, Methanol-d.sub.4) .delta. 7.85 (m, 1H), 7.34-7.22 (m,
2H), 7.22-7.08 (m, 3H), 6.94-6.84 (m, 2H), 4.95-4.85 (m, 1H), 4.79
(m, 1H), 4.46-4.34 (m, 2H), 4.34-4.24 (m, 1H), 4.19 (m, 1H), 3.81
(m, 1H), 2.27 (m, 2H), 2.01 (m, 2H), 1.84-1.68 (m, 1H), 1.62 (m,
1H), 1.30-1.16 (m, 3H). .sup.31P NMR (162 MHz, cd.sub.3od) .delta.
3.70, 3.65. MS m/z=573.0 [M+H].
Example 28. (2S)-isopropyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate (25)
##STR00167##
[0369] The preparation of (2S)-isopropyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate is described below.
Preparation of (2S)-isopropyl
2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)-3-phenylpropanoate
(Compound H)
##STR00168##
[0371] Phenyl dichlorophosphate (718 .mu.L, 4.8 mmol) was dissolved
in 10 mL of anhydrous DCM and stirred under a nitrogen atmosphere
in an ice bath. L-Phenylalanine isopropyl ester hydrochloride (1 g,
4.1 mmol) was added in one portion. Another 10 mL of anhydrous DCM
was added. Triethylamine (736 .mu.L, 5.3 mmol) was added dropwise
and the reaction mixture was stirred for about 30 min. More
triethylamine (736 .mu.L, 5.3 mmol) was then added dropwise and the
reaction mixture was stirred for 30 min. Additional triethylamine
(736 .mu.L, 5.3 mmol) was then added dropwise and the reaction
mixture was stirred for about 15 min. p-Nitrophenol (600 mg, 4.32
mmol) was then added. The ice bath was then removed and the
reaction mixture was allowed to warm to room temperature and
stirred for about 2 h. More p-nitrophenol (50 mg) and triethylamine
(736 .mu.L, 5.3 mmol) were the added and the reaction mixture was
stirred for about 1 h.
[0372] The reaction mixture was then concentrated under reduced
pressure, and was diluted with EtOAc and washed twice with 5%
aqueous citric acid solution, followed with saturated aqueous
sodium chloride solution. The organic layer was dried over
anhydrous sodium sulfate and was concentrated under reduced
pressure. The crude was purified with silica gel column (0-15%
EtOAc in hexanes) to give compound H. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.17 (m, 2H), 7.38-7.13 (m, 10H), 7.13-7.02 (m,
2H), 4.95 (m, 1H), 4.31 (m, 1H), 3.69 (m, 1H), 3.02 (dd, J=6.1, 1.8
Hz, 2H), 1.21-1.08 (m, 6H). .sup.31P NMR (162 MHz, cdcl3) .delta.
-2.96, -2.98. MS m/z=485.0 [M+H].
Preparation of (2S)-isopropyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-3-phe-
nylpropanoate (Compound 25)
##STR00169##
[0374] Compound 1 (58 mg, 0.2 mmol) and compound H (116 mg, 0.24
mmol) were mixed and 2 mL of anhydrous DMF was added. The reaction
mixture was stirred under a nitrogen atmosphere at room
temperature. 1M tBuMgCl in THF (300 .mu.L, 0.3 mmol) was added
dropwise over 3 minutes and the reaction mixture was then stirred
for about 16 h. The reaction mixture was diluted with EtOAc and
washed with 5% aqueous citric acid solution, saturated aqueous
sodium bicarbonate solution and then saturated aqueous sodium
chloride solution. The organic layer was dried over anhydrous
sodium sulfate and concentrated under reduced pressure. The crude
residue was purified with silica gel column (0-5% MeOH in DCM) to
give compound 25. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.84
(m, 1H), 7.27-7.08 (m, 8H), 7.08-6.97 (m, 2H), 6.88 (m, 2H),
4.91-4.84 (m, 1H), 4.74 (m, 1H), 4.26 (m, 1H), 4.19-4.04 (m, 2H),
4.04-3.91 (m, 2H), 2.97 (m, 1H), 2.82 (m, 1H), 1.14 (m, 3H), 1.06
(m, 3H). .sup.31P NMR (162 MHz, CD.sub.3OD) .delta. 3.63, 3.25. MS
m/z=637.0 [M+H].
Example 29. (S)-methyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
Propanoate (26)
##STR00170##
[0376] The preparation of (S)-methyl 2-(((S)-(((2R,3
S,4R,5R)-5-(4-aminopyrrolo[2,1-f][
1,2,4]triazin-7-yl)-5-cyano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(ph-
enoxy)phosphoryl)amino)propanoate is described below.
##STR00171##
[0377] Compound 1 (100 mg, 0.34 mmol) was dissolved in THF (2 mL)
and cooled with an ice water bath. Then 1M t-BuMgCl (0.52 mL, 0.77
mmol) was added dropwise slowly. The resulting mixture was stirred
for about 30 min at room temperature. Then compound I (Prepared
according to WO 2012142085, 219 mg, 0.52 mmol) in THF (2 mL) was
added over 5 min and the resulting mixture was stirred for about 24
h at room temperature. The reaction mixture was then diluted with
EtOAc, cooled under ice-water bath, washed with aq NaHCO.sub.3 (2
mL), washed with brine, dried with sodium sulfate, and concentrated
in vacuo. The resulting mixture was purified by silica gel column
chromatography (MeOH 0 to 20% in DCM) and prep-HPLC (acetonitrile
10 to 80% in water) to give compound 26. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.86 (s, 1H), 7.29 (dd, J=8.6, 7.2 Hz, 2H),
7.21-7.09 (m, 3H), 6.94-6.81 (m, 2H), 4.79 (d, J=5.4 Hz, 1H), 4.38
(ddq, J=10.8, 5.3, 2.7 Hz, 2H), 4.33-4.23 (m, 1H), 4.18 (t, J=5.5
Hz, 1H), 3.86 (dq, J=9.9, 7.1 Hz, 1H), 3.62 (s, 3H), 1.27 (dd,
J=7.2, 1.1 Hz, 3H). MS m/z=533 (M+1).sup.+.
Example 30. (S)-neopentyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
Propanoate (27)
##STR00172##
[0379] The preparation of (S)-neopentyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate is described below.
##STR00173##
[0380] Compound 1 (100 mg, 0.34 mmol) was dissolved in THF (2 mL)
and cooled under ice water bath. Then 1M t-BuMgCl (0.52 mL, 0.77
mmol) was added dropwise slowly. The resulting mixture was stirred
for about 30 min at room temperature. Then compound J (Prepared
according to WO2012075140, 248 mg, 0.52 mmol) was added over about
5 min and the resulting mixture was stirred for about 24 h at room
temperature, diluted with EtOAc, cooled under ice-water bath,
treated with aq NaHCO.sub.3 (2 mL), washed with brine, dried with
sodium sulfate, and concentrated in vacuo. The resulting mixture
was purified by silica gel column chromatography (MeOH 0 to 20% in
DCM) and prep-HPLC (acetonitrile 10 to 80% in water) to give
Compound 27. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.86 (s,
1H), 7.36-7.24 (m, 2H), 7.23-7.10 (m, 3H), 6.96-6.85 (m, 2H), 4.78
(d, J=5.4 Hz, 1H), 4.38 (tdd, J=10.0, 4.9, 2.5 Hz, 2H), 4.32-4.24
(m, 1H), 4.17 (t, J=5.6 Hz, 1H), 3.91 (dq, J=9.8, 7.1 Hz, 1H), 3.81
(d, J=10.5 Hz, 1H), 3.69 (d, J=10.5 Hz, 1H), 1.31 (dd, J=7.2, 1.1
Hz, 3H), 0.89 (s, 9H). MS m/z=589 (M+1).sup.+.
Example 31. (2S)-cyclopentyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
Propanoate (28)
##STR00174##
[0382] The preparation of (2S)-cyclopentyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propan-
oate is described below.
##STR00175##
[0383] Compound 1 (100 mg, 0.34 mmol) was dissolved in THF (2 mL)
and cooled under ice water bath. Then 1M t-BuMgCl (0.52 mL, 0.77
mmol) was added dropwise slowly. The resulting mixture was stirred
for about 30 min at room temperature. Then compound K (Prepared
according to WO2012075140, 247 mg, 0.52 mmol) in THF (2 mL) was
added over about 5 min and the resulting mixture was stirred for
about 24 h at room temperature, diluted with EtOAc, cooled under
ice-water bath, treated with aq NaHCO.sub.3 (2 mL), washed with
brine, dried with sodium sulfate, and concentrated in vacuo. The
resulting mixture was purified by silica gel column chromatography
(MeOH 0 to 20% in DCM) and prep-HPLC (acetonitrile 10 to 80% in
water) to give example 28. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.85 (s, 1H), 7.33-7.22 (m, 2H), 7.14 (tdd, J=7.6, 2.1, 1.1
Hz, 3H), 6.95-6.87 (m, 2H), 5.13-5.00 (m, 1H), 4.78 (d, J=5.4 Hz,
1H), 4.48-4.35 (m, 2H), 4.30 (ddd, J=10.6, 5.7, 3.6 Hz, 1H), 4.19
(t, J=5.4 Hz, 1H), 3.78 (dq, J=9.2, 7.1 Hz, 1H), 1.81 (dtd, J=12.5,
5.9, 2.4 Hz, 2H), 1.74-1.49 (m, 6H), 1.21 (dd, J=7.1, 1.2 Hz, 3H).
MS m/z=587 (M+1).sup.+.
Example 32. (2S)-cyclohexyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)
Propanoate (29)
##STR00176##
##STR00177##
[0385] To a mixture of compound 1 (50 mg, 0.343 mmol), compound M
(Prepared according to US20130143835, 93 mg, 0.209 mmol), and
MgCl.sub.2 (24.5 mg, 0.257 mmol) in DMF (1 mL) was added
diisopropylethylamine (0.075 mL, 0.43 mmol) dropwise over about 5
min at about 0.degree. C. The resulting mixture was stirred at
about 50.degree. C. for about 1 h. The reaction mixture was then
cooled with an ice-water bath, treated with 1M citric acid (0.5
mL), and was purified directly by prep-HPLC (ACN 0 to 70% in water)
to afford compound 29. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.84 (s, 1H), 7.32-7.23 (m, 2H), 7.18-7.10 (m, 3H), 6.93-6.87 (m,
2H), 4.78 (d, J=5.4 Hz, 1H), 4.67 (td, J=8.7, 4.2 Hz, 1H),
4.48-4.35 (m, 2H), 4.30 (ddd, J=10.8, 5.7, 3.7 Hz, 1H), 4.20 (t,
J=5.4 Hz, 1H), 3.88-3.71 (m, 1H), 1.83-1.63 (m, 4H), 1.58-1.46 (m,
1H), 1.46-1.24 (m, 5H), 1.24 (s, 3H). .sup.31P NMR (162 MHz,
CD.sub.3OD) .delta. 3.75. MS m/z=601 (M+1).sup.+.
Example 33. Ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-met-
hylpropanoate (30)
##STR00178##
[0387] The preparation of ethyl 2-(((((2R,3
S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydro-
xytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-methylpropanoa-
te is described below.
Preparation of Ethyl
2-((tert-butoxycarbonyl)amino)-2-methylpropanoate
##STR00179##
[0389] Take up triphenylphosphine (6.18 g, 25.00 mmol) in THF (30
mL). Next charge DIAD (4.92 mL, 25.00 mmol) and stir at room
temperature for 10 min. Dissolve
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (5.08 g,
25.00 mmol) in THF (20 mL) and add to the reaction mixture followed
by the addition of ethanol (2.19 mL, 37.49 mmol). Allow the
reaction to stir at room temperature for about 1 h. The solvents
were removed under reduced pressure and the crude was taken up in
1:1 Et.sub.2O:Hexanes (120 mL). The solid triphenylphosphine oxide
was filtered off and the solvent was removed under reduced
pressure. The crude was taken up in minimal CH.sub.2Cl.sub.2 and
purified by silica gel chromatography 0-50% EtOAc/Hex to afford
ethyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate. .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 4.18 (q, J=7.1 Hz, 2H), 1.49
(s, 6H), 1.43 (s, 9H), 1.27 (t, J=7.1 Hz, 3H).
Preparation of Ethyl 2-amino-2-methylpropanoate Hydrochloride
##STR00180##
[0391] Take up ethyl
2-((tert-butoxycarbonyl)amino)-2-methylpropanoate (2.71 g, 11.72
mmol) in CH.sub.2Cl.sub.2 (25 mL) and slowly add 4N HCl in dioxane
(25 mmol) and stir at room temperature. At 1 h, the reaction was
determined to be complete by TLC. The solvents were removed under
reduced pressure and the crude was coevaporated with Et.sub.2O two
times then placed under high vacuum to afford ethyl
2-amino-2-methylpropanoate hydrochloride. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.70 (s, 3H), 4.18 (q, J=7.1 Hz, 2H), 1.46
(s, 6H), 1.21 (t, J=7.1 Hz, 3H).
Preparation of Ethyl
2-methyl-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate
(Compound N)
##STR00181##
[0393] Take up phenyl dichlorophosphate (0.97 mL, 6.50 mmol) and
ethyl 2-amino-2-methylpropanoate hydrochloride (1.09 g, 6.50 mmol)
in CH.sub.2Cl.sub.2 (50 mL). Cool the reaction mixture to about
0.degree. C. and slowly add TEA (1.75 mL, 12.45 mmol). Remove the
cold bath and allow the reaction mixture to stir at room
temperature. After about 2 h, the addition of the amino acid was
determined to be complete by .sup.31P NMR. Charge p-nitrophenol
(0.860 g, 6.17 mmol) followed by the addition of TEA (0.87 g, 7.69
mmol). Allow the reaction to stir at room temperature. After about
2 h, the reaction was determined to be complete by LCMS. The
reaction was diluted with Et.sub.2O and the TEA.HCl salts were
filtered off The crude was concentrated and purified by silica gel
chromatography (0-50% EtOAc/Hex) to afford compound N. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.37-8.21 (m, 2H), 7.55-7.44 (m,
2H), 7.43-7.33 (m, 2H), 7.30-7.09 (m, 3H), 6.57 (d, J=10.1 Hz, 1H),
3.99 (q, J=7.1 Hz, 2H), 1.39 (s, 6H), 1.08 (t, J=7.1 Hz, 3H).
.sup.31P NMR (162 MHz, DMSO-d.sub.6) .delta. -2.87. LC/MS:
t.sub.R=1.65 min, MS m/z=408.97 [M+1].; LC system: Thermo Accela
1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6.mu.
XB-C18 100A, 50.times.3.00 mm; Solvents: Acetonitrile with 0.1%
formic acid, Water with 0.1% formic acid; Gradient: 0 min-2.4 min
2-100% ACN, 2.4 min-2.80 min 100% ACN, 2.8 min-2.85 min 100%-2%
ACN, 2.85 min-3.0 min 2% ACN at 1.8 mL/min.
Preparation of Ethyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-met-
hylpropanoate (Compound 30)
##STR00182##
[0395] Take up compound 1 (66 mg, 0.23 mmol) in NMP (2.0 mL). Cool
the mixture to about 0.degree. C. and slowly add tBuMgCl (1.0M in
THF, 0.34 mL, 0.34 mmol). Allow the reaction to stir at about
0.degree. C. for about 30 min, then add a solution of compound N
(139 mg, 0.34 mmol) dissolved in THF (1.0 mL). Remove the cold bath
and place the reaction in about 50.degree. C. preheated oil bath.
After about 2 h, the reaction was cooled to room temperature and
quenched with acetic acid and methanol. The crude was concentrated
and purified by reverse phase HPLC without modifier to afford
compound 30. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (m,
3H), 7.31 (q, J=8.1 Hz, 2H), 7.22-7.05 (m, 3H), 6.87 (d, J=4.5,
1H), 6.80 (d, J=4.5 Hz, 1H), 6.27 (d, J=11.7, 1H), 5.81 (d, J=9.7,
1H), 5.35 (d, J=5.6 Hz, 1H), 4.64 (dt, J=9.0, 5.6 Hz, 1H), 4.24 (m,
2H), 4.11 (m, 1H), 4.04-3.90 (m, 3H), 1.39-1.23 (m, 6H), 1.10 (t,
J=7.1, 3H). .sup.31P NMR (162 MHz, DMSO-d.sub.6) .delta. 2.45,
2.41. LC/MS: t.sub.R=1.03 min, MS m/z=561.03 [M+1]; LC system:
Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column:
Kinetex 2.6.mu. XB-C18 100A, 50.times.3.00 mm; Solvents:
Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid;
Gradient: 0 min-2.4 min 2-100% ACN, 2.4 min-2.80 min 100% ACN, 2.8
min-2.85 min 100%-2% ACN, 2.85 min-3.0 min 2% ACN at 1.8
mL/min.
Example 34. Isopropyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-met-
hylpropanoate (31)
##STR00183##
[0397] The preparation of Isopropyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-met-
hylpropanoate is described below.
Preparation of Isopropyl
2-((tert-butoxycarbonyl)amino)-2-methylpropanoate
##STR00184##
[0399] Take up triphenylphosphine (6.17 g, 25.00 mmol) in THF (30
mL). Next charge DIAD (4.92 mL, 25.00 mmol) and stir at room
temperature for about 10 min. Dissolve
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (5.07 g,
25.00 mmol) dissolved in THF (20 mL) and add to the reaction
mixture followed by the addition of isopropanol (1.91 mL, 25.00
mmol). Allow the reaction to stir at room temperature for about 1
h. The solvents were removed under reduced pressure and the crude
was taken up in 1:1 Et.sub.2O:Hexanes (120 mL). The solid
triphenylphosphine oxide was filtered off and the solvent was
removed under reduced pressure. The crude was taken up in minimal
CH.sub.2Cl.sub.2 and purified by silica gel chromatography (0-50%
EtOAc/Hex) to afford isopropyl
2-((tert-butoxycarbonyl)amino)-2-methylpropanoate. .sup.1H NMR (400
MHz, Chloroform-d) .delta. 5.03 (p, J=6.2 Hz, 1H), 1.48 (s, 6H),
1.40 (d, J=6.2 Hz, 9H), 1.24 (d, J=6.3 Hz, 6H).
Preparation of Isopropyl 2-amino-2-methylpropanoate
Hydrochloride
##STR00185##
[0401] Take up isopropyl
2-((tert-butoxycarbonyl)amino)-2-methylpropanoate (4.09 g, 16.67
mmol) in CH.sub.2Cl.sub.2 (50 mL) and slowly add 4N HCl in dioxane
(50 mmol) and stir at room temperature. At about 1 h, the reaction
was determined to be complete by TLC. The solvents were removed
under reduced pressure and the crude was coevaporated with
Et.sub.2O two times then placed under high vacuum to afford
isopropyl 2-amino-2-methylpropanoate hydrochloride. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.61 (s, 3H), 4.96 (p, J=6.2 Hz,
1H), 1.44 (s, 6H), 1.22 (d, J=6.2 Hz, 6H).
Preparation of Isopropyl
2-methyl-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino) propanoate
(Compound 0)
##STR00186##
[0403] Take up phenyl dichlorophosphate (0.83 mL, 5.58 mmol) and
isopropyl 2-amino-2-methylpropanoate hydrochloride (1.01 g, 5.58
mmol) in CH.sub.2Cl.sub.2 (50 mL). Cool the reaction mixture to
0.degree. C. and slowly add TEA (1.61 mL, 11.45 mmol). Remove the
cold bath and allow the reaction mixture to stir at room
temperature. After about 2 h, the addition of the amino acid was
determined to be complete by .sup.31P NMR. Charge p-nitrophenol
(0.74 g, 5.30 mmol) followed by the addition of TEA (0.81, 5.84
mmol). Allow the reaction to stir at room temperature. After about
2 h, the reaction was determined to be complete by LCMS. The
reaction was diluted with Et.sub.2O and the TEA.HCl salts were
filtered off. The crude was concentrated and purified by silica gel
chromatography (0-50% EtOAc/Hex) to afford compound O. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.42-8.19 (m, 2H), 7.55-7.43 (m,
2H), 7.39 (dd, J=8.6, 7.2 Hz, 2H), 7.30-7.12 (m, 3H), 6.53 (d,
J=10.1 Hz, 1H), 4.82 (hept, J=6.3 Hz, 1H), 1.38 (s, 6H), 1.09 (d,
J=6.3, 6H). .sup.31P NMR (162 MHz, DMSO-d.sub.6) .delta. -2.84.
LC/MS: t.sub.R=1.73 min, MS m/z=422.92 [M+1]; LC system: Thermo
Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex
XB-C18 100A, 50.times.3.00 mm; Solvents: Acetonitrile with 0.1%
formic acid, Water with 0.1% formic acid; Gradient: 0 min-2.4 min
2-100% ACN, 2.4 min-2.80 min 100% ACN, 2.8 min-2.85 min 100%-2%
ACN, 2.85 min-3.0 min 2% ACN at 1.8 mL/min.
Preparation of Isopropyl 2-(((((2R,3
S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydro-
xytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)-2-methylpropanoa-
te (Compound 31)
##STR00187##
[0405] Take up compound 1 (66 mg, 0.23 mmol) in NMP (2.0 mL). Cool
the mixture to about 0.degree. C. and slowly add tBuMgCl (1.0M in
THF, 0.57 mL, 0.57 mmol). Allow the reaction to stir at about
0.degree. C. for about 30 min, then add a solution of compound 0
(143 mg, 0.34 mmol) dissolved in THF (1.0 mL). Remove the cold bath
and place the reaction in an about 50.degree. C. preheated oil
bath. After about 2 h, the reaction was cooled to room temperature
and was quenched with acetic acid and methanol. The crude was
concentrated and purified by reverse phase HPLC without modifier to
afford compound 31. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.88 (m, 3H), 7.30 (td, J=8.5, 7.0 Hz, 2H), 7.20-7.04 (m, 3H), 6.87
(d, J=4.5, 1H), 6.80 (d, J=4.5 Hz, 1H), 6.27 (d, 6.1 Hz, 1H), 5.75
(t, J=9.1 Hz, 1H), 5.34 (d, J=5.7 Hz, 1H), 4.81 (p, J=6.3 Hz, 1H),
4.71-4.50 (m, 1H), 4.23 (m, 2H), 4.11 (m, 1H), 4.03-3.83 (m, 1H),
1.37-1.23 (m, 6H), 1.18-1.04 (m, 6H). .sup.31P NMR (162 MHz, DMSO)
.delta. 2.47, 2.43. LC/MS: t.sub.R=1.08 min, MS m/z=575.06 [M+1];
LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet;
Column: Kinetex 2.6.mu. XB-C18 100A, 50.times.3.00 mm; Solvents:
Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid;
Gradient: 0 min-2.4 min 2-100% ACN, 2.4 min-2.80 min 100% ACN, 2.8
min-2.85 min 100%-2% ACN, 2.85 min-3.0 min 2% ACN at 1.8
mL/min.
Example 35. (S)-2-ethylbutyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)
amino)propanoate (32)
##STR00188##
[0407] The preparation of (S)-2-ethylbutyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate is described below.
Preparation of
(3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)dihydrofuran-2(3H)-one
##STR00189##
[0409]
(3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)tetrahydrofuran--
2-ol (15.0 g) was combined with MTBE (60.0 mL), KBr (424.5 mg),
aqueous K.sub.2HPO.sub.4 solution (2.5M, 14.3 mL), and TEMPO (56
mg). This mixture was cooled to about 1.degree. C. Aqueous bleach
solution (7.9% wt.) was slowly charged in portions until complete
consumption of starting material as indicated through a
starch/iodide test. The layers were separated, and the aqueous
layer was extracted with MTBE. The combined organic phase was dried
over MgSO.sub.4 and concentrated under reduced pressure to yield
the product as a solid.
Preparation (4-amino-7-iodopyrrolo[2,1-f][1,2,4]triazine)
##STR00190##
[0411] To a cold solution of 4-aminopyrrolo[2,1-f][1,2,4]-triazine
(10.03 g; 74.8 mmol) in N,N-dimethylformamide (70.27 g),
N-iodosuccinimide (17.01 g; 75.6 mmol) was charged in portions,
while keeping the contents at about 0.degree. C. Upon reaction
completion (about 3 h at about 0.degree. C.), the reaction mixture
was transferred into a 1 M sodium hydroxide aqueous solution (11 g
NaOH and 276 mL water) while keeping the contents at about
20-30.degree. C. The resulting slurry was agitated at about
22.degree. C. for 1.5 h and then filtered. The solids are rinsed
with water (50 mL) and dried at about 50.degree. C. under vacuum to
yield 4-amino-7-iodopyrrolo[2,1-f][1,2,4]triazine as a solid.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 7.90 (s, 1H), 7.78 (br s,
2H), 6.98 (d, J=4.4 Hz, 1H), 6.82 (d, J=4.4 Hz, 1H). .sup.13C NMR
(101 MHz, DMSO-d6) .delta. 155.7, 149.1, 118.8, 118.1, 104.4, 71.9.
MS m/z=260.97 [M+H].
Preparation
(3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-bis(benzyloxy-
)-5-((benzyloxy)methyl)tetrahydrofuran-2-ol via
(4-amino-7-iodopyrrolo[2,1-f][1,2,4]triazine)
##STR00191##
[0413] To a reactor under a nitrogen atmosphere was charged
iodobase 2 (81 g) and THF (1.6 L). The resulting solution was
cooled to about 5.degree. C., and TMSCl (68 g) was charged. PhMgCl
(345 mL, 1.8 M in THF) was then charged slowly while maintaining an
internal temperature at about .ltoreq.5.degree. C. The reaction
mixture was stirred at about 0.degree. C. for 30 min, and then
cooled to about -15.degree. C. iPrMgCl--LiCl (311 mL, 1.1 M in THF)
was charged slowly while maintaining an internal temperature below
about -12.degree. C. After about 10 minutes of stirring at about
-15.degree. C., the reaction mixture was cooled to about
-20.degree. C., and a solution of lactone 1 (130 g) in THF (400 mL)
was charged. The reaction mixture was then agitated at about
-20.degree. C. for about 1 h and quenched with AcOH (57 mL). The
reaction mixture was warmed to about 0.degree. C. and adjusted to
pH 7-8 with aqueous NaHCO.sub.3 (5 wt %, 1300 mL). The reaction
mixture was then diluted with EtOAc (1300 mL), and the organic and
aqueous layers were separated. The organic layer was washed with 1N
HCl (1300 mL), aqueous NaHCO.sub.3 (5 wt %, 1300 mL), and brine
(1300 mL), and then dried over anhydrous Na.sub.2SO.sub.4 and
concentrated to dryness. Purification by silica gel column
chromatography using a gradient consisting of a mixture of MeOH and
EtOAc afforded the product.
Preparation ((2S)-2-ethylbutyl
2-(((perfluorophenoxy)(phenoxy)phosphoryl)amino)propanoate)
(Mixture of Sp and Rp)
##STR00192##
[0415] L-Alanine 2-ethylbutyl ester hydrochloride (5.0 g, 23.84
mmol) was combined with methylene chloride (40 mL), cooled to about
-78.degree. C., and phenyl dichlorophosphate (3.65 mL, 23.84 mmol)
was added. Triethylamine (6.6 mL, 47.68 mmol) was added over about
60 min at about -78.degree. C. and the resulting mixture was
stirred at ambient temperature for 3 h. The reaction mixture was
cooled to about 0.degree. C. and pentafluorophenol (4.4 g, 23.84
mmol) was added. Triethylamine (3.3 mL, 23.84 mmol) was added over
about 60 min. The mixture was stirred for about 3 h at ambient
temperature and concentrated under reduced pressure. The residue
was dissolved in EtOAc, washed with an aqueous sodium carbonate
solution several times, and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography using
a gradient of EtOAc and hexanes (0 to 30%). Product containing
fractions were concentrated under reduced pressure to give
(2S)-2-ethylbutyl
2-(((perfluorophenoxy)(phenoxy)phosphoryl)amino)propanoate as a
solid. .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.41-7.32 (m,
4H), 7.30-7.17 (m, 6H), 4.24-4.16 (m, 1H), 4.13-4.03 (m, 4H),
4.01-3.89 (m, 1H), 1.59-1.42 (m, 8H), 1.40-1.31 (m, 8H), 0.88 (t,
J=7.5 Hz, 12H). .sup.31P NMR (162 MHz, Chloroform-d) .delta. -1.52.
.sup.19F NMR (377 MHz, Chloroform-d) .delta. -153.63,-153.93 (m),
-160.05 (td, J=21.9, 3.6 Hz), -162.65 (qd, J=22.4, 20.5, 4.5 Hz).
MS m/z=496 [M+H].
Preparation ((2S)-2-ethylbutyl
2-(((perfluorophenoxy)(phenoxy)phosphoryl)amino)propanoate)
##STR00193##
[0417] L-alanine-2-ethylbutylester hydrochloride (40.10 g, 0.191
mmol) was dissolved in dichloromethane (533 g) and the solution was
cooled with stirring to about -15.degree. C. under N.sub.2(g).
Phenyl dichlorophosphate (40.32 g, 0.191 mol) was added followed by
slow addition of triethylamine (41.58 g, 0.411 mmol) and the
reaction mixture was stirred at about -15.degree. C. for about 1.5
h. Pentafluorophenol (35.14 g, 0.191 mol) was added, followed by
triethylamine (19.23 g, 0.190 mol) and the reaction mixture was
stirred for about 2 h. The reaction mixture was warmed to about
0.degree. C. and 0.5 M HCl (279.19 g) was added. The mixture was
warmed to about 22.degree. C. and the organic layer was separated
and washed with 5% KHCO.sub.3 aqueous solution (281 g), then water
(281 g). An aliquot of the organic layer (453.10 g of the 604.30 g
solution) was concentrated to about 120 mL volume, isopropyl
acetate (157 g) was added and the solution was concentrated to
dryness. The residue was dissolved in isopropyl acetate (158 g).
The resulting solution was concentrated to about 120 mL volume and
the temperature was adjusted to about 45.degree. C. n-Heptane (165
g) was added and the mixture was cooled to 22.degree. C. over about
1 h. n-Heptane (167 g) was added and the mixture was cooled to
about 0.degree. C. Triethylamine (2.90 g, 0.0287 mol) was added and
the mixture was stirred at 0.degree. C. for about 17 h. The mixture
was filtered, the solids were rinsed with n-heptane (145 g) and the
solids were dried under vacuum at about 40.degree. C. for about 15
h to provide 2-ethylbutyl
((S)-(penthafluorophenoxy)(phenoxy)phosphoryl)-L-alaninate.
Preparation 2-ethylbutyl
((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate
##STR00194##
[0419] A slurry of L-alanine-2-ethylbutylester hydrochloride (20.08
g, 95.8 mmol) and isopropyl acetate (174 g) was cooled with
stirring to about -20.degree. C.). Phenyl dichlorophosphate (20.37
g, 96.5 mmol) was added, followed by slow addition of triethyl
amine (20.97 g, 207.2 mmol) and the mixture was stirred at about
-20.degree. C. for about 1 h. 4-Nitrophenol (13.23 g, 95.1 mmol)
was added, followed by slow addition of triethylamine (10.01 g,
98.8 mmol) and the reaction mixture was stirred for about 1.5 h.
The reaction mixture was warmed to about 0.degree. C. and 0.5 M HCl
(140 g) was added. The organic layer was separated and washed with
5% Na.sub.2CO.sub.3 (2.times.100 g) and 10% NaCl (2.times.100 g).
The organic layer was then concentrated to about 80 mL volume and
isopropylacetate (4 g) was added, followed by n-heptane (110 g).
Product seed crystals (0.100 g) were added followed by a second
portion of n-heptane (110 g) and the mixture was cooled to about
0.degree. C. 1,8-Diazabicycloundec-7-ene (1.49 g, 9.79 mmol) was
added and the mixture was stirred at about 0.degree. C. for about
21 h. The resultant solids were filtered and washed first with
n-heptane (61 g) and then with H.sub.2O (2.times.100 g). The solids
were stirred with H.sub.2O (200 g) for about 1.5 h, filtered, and
rinsed with H.sub.2O (3.times.100 g), then n-heptane (61 g). The
obtained solids were dried under vacuum at about 40.degree. C. for
about 19 h to provide 2-ethylbutyl
((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate.
Preparation of Title Compound (Mixture of Sp and Rp)
##STR00195##
[0421] The nucleoside (29 mg, 0.1 mmol) and the phosphonamide (60
mg, 0.12 mmol) and N,N-dimethylformamide (2 mL) were combined at
ambient temperature. Tert-Butyl magnesium chloride (1M in THF, 0.15
mL) was slowly added. After about 1 h, the reaction was diluted
with ethyl acetate, washed with aqueous citric acid solution (5%
wt.), aqueous saturated NaHCO.sub.3 solution and saturated brine
solution. The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography using a gradient of methanol and
CH.sub.2Cl.sub.2 (0 to 5%). Product containing fractions were
concentrated under reduced pressure to provide the product.
Preparation of
(3aR,4R,6R,6aR)-4-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-6-(hydroxyme-
thyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carbonitrile
##STR00196##
[0423] To a mixture of
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile (5.8 g, 0.02 mol),
2,2-dimethoxypropane (11.59 mL, 0.09 mol) and acetone (145 mL) at
ambient temperature was added sulfuric acid (18M, 1.44 mL). The
mixture was warmed to about 45.degree. C. After about 30 min, the
mixture was cooled to ambient temperature and sodium bicarbonate
(5.8 g) and water 5.8 mL) were added. After 15 min, the mixture was
concentrated under reduced pressure. The residue was taken up in
ethyl acetate (150 mL) and water (50 mL). The aqueous layer was
extracted with ethyl acetate (2.times.50 mL). The combined organic
phase was dried over sodium sulfate and concentrated under reduced
pressure to give crude
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile. .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.84 (s, 1H), 6.93 (d, J=4.6 Hz, 1H), 6.89
(d, J=4.6 Hz, 1H), 5.40 (d, J=6.7 Hz, 1H), 5.00 (dd, J=6.7, 3.3 Hz,
1H), 4.48-4.40 (m, 1H), 3.81-3.72 (m, 2H), 1.71 (s, 3H), 1.40 (s,
3H). MS m/z=332.23 [M+1].
Preparation of
(3aR,4R,6R,6aR)-4-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-6-(hydroxyme-
thyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carbonitrile
TsOH Salt
##STR00197##
[0425] To a mixture of
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile (5.0 g, 17.2 mmol,
1.0 equiv.), 2,2-dimethoxypropane (10.5 mL, 86 mmol, 5.0 equiv.)
and acetone (25 mL) at ambient temperature was added
p-tolylsulfonic acid (3.59 g, 1.1 equiv.). The mixture was stirred
at ambient temperature. After about 30 min, isopropyl acetate (25
mL) was added over about one hour. The resulting slurry was
filtered and rinsed with 2:1 heptane:isopropyl acetate (25 ml). The
product was dried under vacuum at about 40.degree. C.
Preparation of
(3aR,4R,6R,6aR)-4-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-6-(hydroxyme-
thyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carbonitrile
##STR00198##
[0427] To a mixture of
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile (5 g, 17.2 mmol,
1.0 equiv.), 2,2-dimethoxypropane (10.5 mL, 86 mmol, 5.0 equiv.)
and acetone (25 mL) at ambient temperature was added
p-tolylsulfonic acide (3.59 g, 1.1 equiv.). The mixture was stirred
at ambient temperature. After 30 min, isopropyl acetate (25 mL) was
added over one hour. The resulting slurry was filtered and rinsed
with 2:1 heptane:isopropyl acetate (25 ml). The product was dried
under vacuum at 40.degree. C. The isolated solid was added to a
reactor and 5% K.sub.2CO.sub.3 solution (50 ml) and ethyl acetate
(50 mL) were added. The layers were separated, and the aqueous
layer washed with ethyl acetate (25 ml). The combined organic
layers were washed with water (25 ml), then concentrated to ca.25
ml. The reactor was refilled with isopropyl acetate (25 ml) and
concentrated to ca. 25 ml. The reactor was again refilled with
isopropyl acetate (25 ml) and concentrated to 25 ml. The resulting
solution was seeded, producing a thick slurry. To this was added
heptane (25 ml) over one hour. The resulting slurry was filtered
and rinsed with 2:1 heptane:isopropyl acetate (25 ml). The product
was dried under vacuum at 40.degree. C. 0
(2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy--
5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile. .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.84 (s, 1H), 6.93 (d, J=4.6 Hz, 1H), 6.89
(d, J=4.6 Hz, 1H), 5.40 (d, J=6.7 Hz, 1H), 5.00 (dd, J=6.7, 3.3 Hz,
1H), 4.48-4.40 (m, 1H), 3.81-3.72 (m, 2H), 1.71 (s, 3H), 1.40 (s,
3H). MS m/z=332.23 [M+1].
Preparation of (2S)-2-ethylbutyl
2-(((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano--
3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propan-
oate
##STR00199##
[0429] Acetonitrile (100 mL) was combined with (2S)-2-ethylbutyl
nitrophenoxy)(phenoxy)phosphoryl)-amino)propanoate (9.6 g, 21.31
mmol), the substrate alcohol (6.6 g, 0.02 mol),), magnesium
chloride ((1.9 g, 19.91 mmol) at ambient temperature. The mixture
was agitated for about 15 min and N,N-diisopropylethylamine (8.67
mL, 49.78 mmol) was added. After about 4 h, the reaction was
diluted with ethyl acetate (100 mL), cooled to about 0.degree. C.
and combined with aqueous citric acid solution (5% wt., 100 mL).
The organic phase was washed with aqueous citric acid solution (5%
wt., 100 mL) and aqueous saturated ammonium chloride solution (40
mL), aqueous potassium carbonate solution (10% wt., 2.times.100
mL), and aqueous saturated brine solution (100 mL). The organic
phase was dried with sodium sulfate and concentrated under reduced
pressure to provide crude product. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.86 (s, 1H), 7.31-7.22 (m, 2H), 7.17-7.09 (m,
3H), 6.93-6.84 (m, 2H), 5.34 (d, J=6.7 Hz, 1H), 4.98 (dd, J=6.6,
3.5 Hz, 1H), 4.59-4.50 (m, 1H), 4.36-4.22 (m, 2H), 4.02 (dd,
J=10.9, 5.7 Hz, 1H), 3.91 (dd, J=10.9, 5.7 Hz, 1H), 3.83 (dq,
J=9.7, 7.1 Hz, 1H), 1.70 (s, 3H), 1.50-1.41 (m, 1H), 1.39 (s, 3H),
1.36-1.21 (m, 7H), 0.86 (t, J=7.4 Hz, 6H). MS m/z=643.21 [M+1].
Preparation of (S)-2-ethylbutyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate (Compound 32)
##STR00200##
[0431] The crude acetonide (12.85 g) was combined with
tetrahydrofuran (50 mL) and concentrated under reduced pressure.
The residue was taken up in tetrahydrofuran (100 mL), cooled to
about 0.degree. C. and concentrated HCl (20 mL) was slowly added.
The mixture was allowed to warm to ambient temperature. After
consumption of the starting acetonide as indicated by HPLC
analysis, water (100 mL) was added followed by aqueous saturated
sodium bicarbonate solution (200 mL). The mixture was extracted
with ethyl acetate (100 mL), the organic phase washed with aqueous
saturated brine solution (50 mL), dried over sodium sulfated and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography using a gradient of methanol and
ethyl acetate (0 to 20%). Product containing fractions were
concentrated under reduced pressure to provide the product.
Preparation of (S)-2-ethylbutyl
2-(((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cy-
ano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)pr-
opanoate (Compound 32)
##STR00201##
[0433] To a vial containing (S)-2-ethylbutyl
2-(((S)-(((3aR,4R,6R,6aR)-6-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-6--
cyano-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)(phenoxy)p-
hosphoryl)amino)propanoate (30 mg, 0.05 mmol) was added an 80%
aqueous formic acid solution (1.5 mL). After 18 h at about
20.degree. C. complete conversion was confirmed by HPLC and LC-MS.
MS