Note: Descriptions are shown in the official language in which they were submitted.
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PRODRUGS OF PHOSPHOROUS-CONTAINING PHARMACEUTICALS
Technical Field
This invention relates to the field of phosphorous-containing pharmaceuticals,
including phosphonates, phosphinates and phosphates and in particular to
prodrugs
of such pharmaceuticals. The invention provides novel pharmaceutical compounds
and compositions, methods for their preparation and pharmaceutical methods
employing them.
to Background Art
Many phosphorous-containing pharmaceuticals have not reached their full
clinical
potential due to difficulties in administering them, particularly via the oral
route. Bis-
phosphonate bone resorption agents such as alendronate, for example, have oral
bioavailabilities of just a few percent and adsorption is diminished by food
fruit juice
15 coffee, dairy products etc. This poor bioavailability is exacerbated by the
caustic
nature of the drug which leads to irritation of the gastric walls, resulting
in an
administration regime calling for a fasted stomach and the patient standing
for half an
hour after ingestion to ensure transport of the drug past the gastric region.
20 A significant amount of work has been done to attempt to solve the
administration
difficulties by optimising the formulation of phosphorous-containing drugs.
Merck
for example describe various formulations of the bis-phosphonate alendronate,
such
as a pH buffered formulation in W098 14196, an effervescent formulation in
W097
44017, the monohydrate and disodium in W096 39410, the anhydrate in W096
25 39149, a liquid formulation with complexing agent in W095 33755 and calcium
salts
in EP 449 405. To our knowledge, however, none of these conventional
formulation
approaches has produced a clinically succesful administration form.
Merck and others have also explored prodrugs of bis-phosphonate bone
resorption
3o agents in an attempt to improve the clinical utility of these drugs. For
example,
Leiras OY's US 5,376,649 describes esters of halogenated bis-phosphonates of
the
clodronate type, wherein the prodrug moiety comprises an alkyl, alkenyl,
alkynyl,
aryl, araikyl and silyl. No improvements in bioavailability are reported.
Leo's US
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2
4,732,998, US 4,870,063 and W086 00902 describe prodrugs of certain non-
hydroxylated bis-phosphonates. In particular the pivaloyloxymethyl and
acetoxyrnethyl tetraesters are prepared (although no improvements in
bioavailability
are reported) and certain other esters postulated, such as the C3-C6
alkanoyloxymethyl, C4 C, 1-(alkanoyloxy)ethyl, CS-C8 1-methyl-1-
(alkanoyloxy)ethyl, C4-C, 1-(alkoxycarbonyloxy)ethyl, C3-C6
alkoxycarbonyloxymethyl, CQ-C, 1-alkoxycarbonyloxy)ethyl, CS-Cg 1-methyl-1-
(alkoxycarbonyloxy)ethyl, 3-phthalidyl, 4-crotonolactonyl, y-butyrolacton-4-
yl, (2-
oxo-1,3-dioxolen-4-yl)methyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl, (5-
phenyl-
2-oxo-1,3-dioxolen-4-yl)methyl, dialkylaminoalkyl, acetonyl and methoxymethyl.
Merck's US 5,227,506 describes acyloxymethyl esters of a number of bis-
phosphonates, notable alendronate, wherein the acyl component is C,-C,Z
alkanoyl.
As with the Leo patents above the preferred prodrug is pivaloylxoymethyl,
although
no bioavailability data is provided.
Other phosphorous-containing drugs where attempts to improve bioavailability
shortcomings have employed prodrugs include the antiviral agent foscarnet
(phosphonoformate). Astra's W098 16537 and W09825938 describe
2o monosaccharide and cyclic esters of foscarnet. Boehringer's W09722368,
Hostetler's US 5,696,277 and W094/13682, UC's W096 15132 (inter alia) describe
lipid or glycerol esters of foscarnet. Iyer et al, Tett. Letters 30 No 51,
7141-7144
(1989) describe acyloxyalkyl esters, wherein the acyl component is C,-C4
alkanoyl.
Glazier's W091/19721 describes acyloxybenzyloxy esters. Walker et al, Int J
Pharmaceutics 104 (1994)157-167 describes foscarnet-tyrosine conjugates
wherein
the hydroxy group on the amino acid side chain is esterified to the
phosphonate.
These compounds are not however orally administered prodrugs, but rather are
designed to penetrate the blood brain barner following iv administration.
3o A still further group of pharmaceuticals where prodrugs have been proposed
to
attempt to overcome bioavailability shortcomings are the phosphonate
nucleoside
analogues, such as those disclosed in Gilead's W098 04569 and an extensive
series
of patents in the name Rega Stichting/Institute of Organic Chemistry &
Biochemistry
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WO 99/51613 PCT/SE99/00528
of the Academy of Sciences of the Czech Republic including EP 481214. However,
to our knowledge, the only such phosphonates to show clinical practicality are
the
now marketed bis-POM and bis-POC adefovir (bis-pivoxil and bis-
isopropyloxycarbonyloxymethyl esters, respectively). WO 95/01363 also shows a
form of phosphonate nucleoside analogue.
Monophosphate esters of monophosphorylated nucleoside analogues are desclosed
in
EP 763049 (glyceryl diethers), WO 95 32984 (lipid esters), US 5484911 (ether
lipids)WO 98 51692 (substituted benzyl esters) WO 96 33201 (alkylthioacyl
esters)
Glazier's WO 91/19721 (above) and US 5 627 165 and Texas Uni's WO 90/08155
purport to provide prodrugs of diverse phosphorous-containing pharmaceuticals.
Bodor's WO 92/00988 employs a phosphonate prodrug of non-phosphorous
pharmaceuticals to target compounds to the brain.
Brief Description of the Invention
A first aspect of the invention provides pharmaceutical compound with the
formula:
Drug-P(=O)-O-Linker (-R2')k -Rz
where Drug-P(=O)-O- is the residue of a drug comprising a phosphonate,
phosphinate or phosphoryl function,
RZ and RZ' (if present) are independently the acyl residue of an aliphatic
amino acid;
Linker is an at least difunctional moiety comprising a first function ester-
bonded to
the phosphonate, phosphinate or phosphoryl function spaced from a second
function
ester-bonded to Rz; and
k is 1 or zero.
The enzymatic and/or chemical cleavage of the compounds of the present
invention
occurs in such a manner that the parent drug is released and the moiety or
moieties
split off remain non-toxic or are metabolized so that non-toxic or acceptable
amounts
of metabolic products are produced.The present compounds thus modify the in
vivo
availability of the parent compound compared to what would be the case if the
parent
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4
compound was to be administered itself. For instance the prodrugs of the
invention
may give higher bioavailabities, varied bioavailability kinetics or
bioavailabilities
with a decreased interpersonal spread.
Representative phosphorous-containing drugs amenable to the prodrugs of the
present invention include phosfestrol, (E)-(a,(3-diethyl-4,4'-
stilbenylen)bis(dihydrogenphosphate), nucleoside analogue phosphonates,
nucleotide
analogue mono-, di or triphosphates, phosphonoformic acid, phosphonoacetic
acid,
bis phosphonate bone metabolism agents, fosinoprilate, (3-phosphonocarboxylic
acid
to farnesyl protein transferase inhbitors, a-phosphonosulfonate squalene
synthase
inhibitors, phosphonomethylamine neutral endopeptidase (24.11 ) inhibitors.
Favoured -linker (-RZ')k - R2 structures include those of the formulae:
R4~ p RaR
R2- O - ()ql ()qr-T -~ o
R4L' R4R 1 I a
or
R4R
R2 - O - ()ql Ring - ()qr-T --~ o
R4R Ilb
1$
where
Rz is the acyl residue of an aliphatic amino acid,
R4Land R4~' are independently H, hydroxymethyl, C,_3 alkyl, C3-Cbcycloalkyl,
C,-
2o C,alkyl-C,-C6cycloalkyl, phenyl or benzyl,
R4R and R4R' are independently H, C,_3 alkyl or phenyl,
ql is 0-3, qr is 0-3,
T is a bond, -NR4- or -O-
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R4 is H or C,_,alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle.
Additional -linker (-RZ')k-RZ structures and intermediates for their
application, such
5 as those of the formula:
R2-O O
O
R2'- O
where RZ and Rz' are as described above are disclosed and claimed in our co-
pending
application PCT/SE99/00194, the contents of which are hereby incorporated by
reference.
Further linker-RZ structures include:
R2~ O'\
such as alanyloxymethyl-, valyloxymethyl-, leucyloxymethyl- t-leucyloxymethyl-
ar isoleucyloxymethyl, especially valyloxymethyl-.
Where the Drug comprises a phosphonate or phosphoryl function, or a plurality
of
such functions, the compound of the invention may comprise a plurality of
-linker (-RZ')~ - Rz structures, preferably identical to each other for ease
of synthesis.
Preferred values for R4Rand R4R' are hydrogen for ease of synthesis, although
compounds wherein one of R4Rand R4R' is hydrogen and the other methyl have the
advantage of releasing the innocuous biproduct ethanol upon degradation.
Representative values for R4L and R4L' include methyl, hydrogen;
hydroxymethyl,
methyl; or ethyl, ethyl, respectively. A particularly preferred value is
methyl, methyl.
Favoured values for ql and qr, respectively, include
1,0;
2,0;
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6
3,0; or
4,0;
particularly 1,0 when the -linker (-Rz')k-Rz structure has the formula IIa.
Alternative convenient values for ql and qr, respectively, include
1,1;
2,1;
3,1;
4,1; or
l0 2,2.
Where the -linker (-RZ')k-RZ structure has the formula IIb, that is includes a
-ring-
moiety an alternative convenient configuration has ql and qr both 0.
The -ring- moiety in the above depicted -linker (-RZ')k-RZ structure may
comprise
furyl, thienyl, pyranyl, pynrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl,
pyrazolinyl,
pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl,
pyrazinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl,
isoxazolyl,
isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl,
isothiazolidinyl,
and the like or bicyclic rings especially of the above fused to a phenyl ring
such as
2o indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl,
benzoxazolyl,
benzothienyl etc. The carbo or heterocyclic ring may be bonded via a carbon or
via a
hetero atom, for instance a nitrogen atom, such as N-piperidyl, N-morpholinyl
etc.
The -ring- moiety is conveniently phenyl, furyl, pyridyl, cyclobutyl,
cyclopentyl or
cyclohexyl.
The -ring- moiety can be optionally substituted with one to three substituents
such as
halo, amino, mercapto, oxo, vitro, NHC,-C6 alkyl, N(C,-C6 alkyl)2, C,-C6
alkyl, C,-C6
alkenyl, C,-C6 alkynyl, C,-C6 alkanoyl, C,-C6 alkoxy, thioC,-C6 alkyl, thioC,-
C6
alkoxy, hydroxy, hydroxyC,-C6 alkyl, haloC,=C6 alkyl, aminoC,-C6 alkyl, C,-C6
alkyl,
3ti cyano, carboxyl, carbalkoxy, carboxamide, carbamoyl, sulfonylamide,
benzyloxy,
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morpholyl-C,-C6 alkyloxy, a monocyclic carbo- or heterocycle, as defined
above, a
optional substituent comprises an additional group RZ ( )q,-.
The aliphatic amino acid RZ is derived from D- or L- glycine, alanine, v ine,
leucine,
tent-leucine and isoleucine. Preferably the or each RZ is derived fro -valine
to
ensure nature identical breakdown products.
1o The currently prefer ed value for T is a bond.
Representative phosphorous-containing drugs incl a phosfestrol, (E)-(a,(3-
diethyl-
4,4'-stilbenylen)bis(dihydrogenphosphate) and ytostatic metabolites such as
phosphorylated nucleoside anlaogues such FLG, cytarabin or gemcitabin.
Representative drugs comprising a ply6sphonate function include antiviral
nucleoside
or nucleotide analogues such as P EA, HPMPC, PMPA and the like or phosphates
such as the monophosphates, ' hosphate or triphospates of those nucleoside
analogues which require p sphorylation for activity, such as gemcitabine, ACV,
2o AZT, ddI, ddC, PCV, V, H2G, BVDU, FMAU, 3TC, FTC etc. Certain mixed
amino acid/fatty aci acyloxyalkylphosphonates are proposed in our copending
application PCT 97 001903 and it should be thus appreciated that the prodrugs
of
the present in ention are fatty acyl-free and/or apply the novel linkers
defined herein
in the nho honate nucleotide field.
Taking the phosphonate antivirals adefovir and cidovir as examples, prodrugs
of the
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carbo- or heterocyclic group spaced by alkyl, such as C1_3 alkylaryl, etc. A
favoured
optional substituent comprises an additional group R2-( )q,-.
The aliphatic anuno acid R2 is derived from D- or L- glycine, alanine, valine,
leucine,
tent-leucine and isoleucine. Preferably the or each R2 is derived from L-
valine to
ensure nature identical breakdown products.
The currently preferred value for T is a bond.
Representative phosphorous-containing drugs include phosfestrol, (E)-(a,,(3-
diethyl-
4,4'-stilbenylen)bis(dihydrogenphosphate) and cytostatic metabolites such as
phosphorylated nucleoside anlaogues such as FLG, cytarabin or gemcitabin.
Representative drugs comprising a phosphonate function include antiviral
nucleoside
or nucleotide analogues such as PMEA, HPMPC, PMPA and the like or phosphates
such as the monophosphates, diphosphate or triphospates of those nucleoside
analogues which require phosphorylation for activity, such as gemcitabine,
ACV,
AZT, ddI, ddC, PCV, GCV, H2G, BVDU, FMAU, 3TC, FTC etc. Certain mixed
amino acid/fatty acid acyloxyalkylphosphonates are proposed in our copending
application PCT SE97 001903 (W098 21223) and it should be thus appreciated
that
the prodrugs of the present invention are fatty acyl-free and/or apply the
novel linkers
defined herein in the phosphonate nucleotide field.
Taking the phosphonate antivirals adefovir and cidovir as examples, prodrugs
of the
invention can be applied as shown in Formula PF2:
AMENDED SHEET
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8
R4~ O R4R O Base
RZ-O- ()Gi 04r-T-~O O_~P~O
R RaR~ Rf3
Rf4
or
O RaR O Base
R2-O- ()qi -ring- ()Gr-T~-O O-P O
R4R~ Rf3
Rf4
where
RZ is the acyl residue of an aliphatic amino acid,
R4~and R4L' are independently H, C,_3 alkyl, hydroxymethyl, C3-6cycloalkyl, C,-
3alkyl-C,C6cycloalkyl phenyl or benzyl,
R4R and R4R' are independently H, C,_3 alkyl or phenyl
ql is 0-3, qr is 0-3,
T is a bond, -NR4- or -O-
to R4 is H or C,_,alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
base is a natural or unnatural nucleotide base, especially guanine, adenine or
cytosine,
Rf3 is H or a further structure of the formula IIa or IIb and Rf4 is H or
CHZOH.
Currently favoured values in formula PF2 include: R3R and R3R' are preferably
H
and/or R,L and R3L' are preferably ethyl or especially methyl. T is preferably
-O- or
more preferably a bond. Preferably qr is 1 or more preferably 0 and q1 is 0 or
more
preferably 1.
Thus a preferred group of phosphonate antivirals within the scope of the
invention
include:
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9-[2-phosphonomethoxy)ethyl]adenine, mono(2-methyl-2-(L-valyloxymethyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl)adenine, mono(2-methyl-2-(L-valyloxy)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono(2-(L-valyloxy)-3-methyl-(S)-(+)-
butyryloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono(2-(-L-valyloxy)-2-phenyl-DL-
acetyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono((1,3-di-valyloxy)propyl-2-
oxycarbonyloxy methyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono(2-L-valyloxy)-DL-
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono-(5-(L-vaiyloxy)-2,2-
dimethylvaleryloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono-((2-(L-valyloxy)-ethoxycarbonyloxy)
methyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono [4-( L-valyloxy)-butanoyloxymethyl)
ester,
9-[2-phosphonomethoxy)ethyl)adenine, mono-(4-(L-valyloxy) benzoyloxymethyl)
ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono-(3-(3,4-di-(L-valyloxy) phenyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl)adenine, mono-(2-methyl-1-(L-valyloxy)-2-
propoxycarbonyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine, mono-(4-N-
valyloxy)cyclohexanoyloxymethyl) ester
9-[2-phosphonomethoxy)ethyl]adenine, mono-(1-valyloxy-2-methylpropane-2-
aminocarbonyloxymethyl) ester
9-[2-phosphonomethoxy)ethyl]adenine, mono-(1-(2-L-valyloxyethyl)-6-oxo-1,6-
3o dihydro-pyridine-3-carbonyloxymethyl) ester
9-[2-phosphonomethoxy)ethyl]adenine, mono-((1,3-di-(valyloxy)propyl-2-
oxycarbonyloxy methyl) ester
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9-[2-phosphonomethoxy)ethyl]adenine, mono-(2-hydroxymethyl-2-
methylpropionyloxymethyl) ester
9-[2-phosphonomethoxy)ethylJadenine, mono-(2-(L-valyloxymethyl)-2-ethyl
butyroyloxymethyl) ester
5 9-[2-phosphonomethoxy)ethyl]adenine, mono-(3-(L-valyloxy)-2-methyl-
propionyloxymethyl) ester
9-[2-phosphonomethoxy)ethylJadenine, mono-(3-(L-valyloxy)-2-methyl, 2-
hydroxymethylpropionyloxymethyl) ester
the corresponding bis esters and pharmaceutically acceptable salts thereof. A
further
10 preferred group comprises the corresponding derivatives of PMPA and HPMPC.
A particularly preferred group comprises.
9-[2-phosphonomethoxy)ethylJadenine di-(2-methyl-2-(D-valyloxymethyl)
propionyloxymethyl) ester
9-[2-phosphonomethoxy)ethyl]adenine mono-(2-methyl-2-(D-valyloxymethyl)
propionyloxyrnethyl) ester
9-[2-phosphonomethoxy)ethylJadenine di-(2-methyl-2-(L-valyloxy)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethylJadenine mono-(2-methyl-2-(L-valyloxy)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethylJadenine di-(2-methyl-2-(D-valyloxy)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethylJadenine mono-(2-methyl-2-(D-valyloxy)
propionyloxymethyl) ester and especially
9-[2-phosphonomethoxy)ethyl]adenine di-(2-methyl-2-(L-valyloxymethyl)
propionyloxymethyl) ester and
9-[2-phosphonomethoxy)ethyl]adenine mono-(2-methyl-2-(L-valyloxymethyl)
propionyloxymethyl) ester;
and the corresponding derivatives of PMPA and HPMPC.
A further convenient group comprises
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9-[2-phosphonomethoxy)ethyl]adenine mono -(2-methyl-2-(L-isoleucyloxymethyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine mono -(2-methyl-2-(L-leucyloxymethyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine mono -(2-methyl-2-(L-t-leucyloxymethyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine mono -(2-methyl-2-(L-alanyloxymethyl)
propionyloxymethyl) ester,
9-[2-phosphonomethoxy)ethyl]adenine mono (2-methyl-2-(L-glycyloxymethyl)
to propionyloxymethyl) ester,
the con:esponding bis esters and the corresponding derivatives of PMPA and
HPMPC.
A further group of useful compounds comprises:
i5 9-[2-phosphonomethoxy)ethyl]adenine mono-L-valyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine di-L-valyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine mono-L-leucyloxymethyl ester,
9-[2-phosphonomethoxy}ethyl]adenine di-L-leucyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine mono-L-isoleucyloxymethyl ester,
2o 9-[2-phosphonomethoxy)ethyl]adenine di-L-isoleucyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine mono-L-t-leucyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine di-L-t-leucyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine mono-L-alanyloxymethyl ester,
9-[2-phosphonomethoxy)ethyl]adenine di-L-alanyloxymethyl ester,
25 and the corresponding derivatives of HPMPC and PMPA.
This aspect of the invention further provides a method for the treatment or
prophylaxis of virus infections comprising the oral administration of an
effective
amount of a compound of the invention to a mammal (including humans) in need
3o thereof. Viral infections include herpesvirus infections such as HSV-1, HSV-
2, VZV,
CMV, HHV6, HHV8 and retroviruses such as HIV-1 and HN-2. The invention
further provides the use of the compounds defined above in medicine and the
use of
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12
these compounds in the preparation of a medicament for the prophylaxis or
treatment
of viral infections.
A further group of phosphorous containing antivirals amenable to the invention
include foscarnet (phosphonoformate) and PAA (phosphonoacetate). Taking
foscarnet as an example:
O O~~
HO-P-u-OH
I
O
H
it will be apparent that aside from conventional pharmaceutically acceptable
esters
to applied to the carboxy function, one or two -linker(-Rz')k Rz structures,
preferably
those of formula IIa or IIb can be applied to the phosphonate function to
define
compounds such as
O
O O
O~O-P-~OH
l
O
H
Thus a preferred group of compounds comprises foscarnet derivatives of the
formula
PF 1:
R4~ O R4R O O
R2-O- ()qi ()qr-T--u--O O-P-~O-Rf2
R4~ R4R
Rf1
or
R4R ~ O
R2-O-Oq~ Ring-()qr-T-11-O -P-ll-O-Rf2
O
_ R4R
Rf1
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13
where
RZ is the acyl residue of an aliphatic amino acid,
S R4Land R4L' are independently H, C,_3 alkyl, hydroxymethyl; C3-bcycloalkyl,
C,-
,alkyl-C,Cbcycloalkyl phenyl or benzyl,
R4R and R4R' are independently H, C,_3 alkyl or phenyl
ql is 0-3, qr is 0-3,
T is a bond, -NR4 or -O-
to R4 is H or C,_3alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
and Rfl is H, a further ester of formula IIa or IIb and Rf2 is H, a further
linker(-RZ')e-
RZ ester or a conventional pharmaceutically acceptable ester, including
acyloxyalkyl
esters.
Currently favoured values in Formula PF1 include: R4R and R4R' are preferably
H
and/or R4L and R4L' are preferably ethyl or especially methyl. T is preferably
-O- or
more preferably a bond. Preferably qr is 1 or more preferably 0 and ql is 0 or
preferably 1. If Rfl is a further ester it is convenient if it is identical to
the other
linker{RZ')k-RZ moiety. Conventional pharmaceutically acceptable esters for
Rf2
include the methyl, ethyl and isopropyl esters or alternatively a similar
structure to
the linker(-RZ')k-RZ moiety as envisaged in our copending PCT/SE99/00194.
A favoured group of compounds within formula PF1 include:
phosphonoformic acid, mono(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl)
ester,
phosphonoformic acid, mono(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
phosphonoformic acid, mono(2-(L-valyloxy)-3-methyl-(S)-(+)-butyryloxymethyl)
ester,
3o phosphonoformic acid, mono(2-(-L-valyloxy)-2-phenyl-DL-acetyloxymethyl)
ester,
phosphonoformic acid, mono((1,3-di-valyloxy)propyl-2-oxycarbonyloxy methyl)
ester,
phosphonoformic acid, mono(2-L-valyloxy)-DL-propionyloxymethyl) ester,
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phosphonoformic acid, mono-(5-(L-valyloxy)-2,2-dimethylvaleryloxymethyl)
ester,
phosphonoformic acid, mono-((2-(L-valyloxy)-ethoxycarbonyloxy) methyl) ester,
phosphonoformic acid, mono [4-( L-valyloxy)-butanoyloxymethyl] ester,
phosphonoformic acid, mono-(4-(L-valyloxy) benzoyloxymethyl) ester,
phosphonoformic acid, mono-(3-(3,4-di-(L-valyloxy) phenyl) propionyloxymethyl)
ester,
phosphonoformic acid, mono- (2-methyl-1-(L-valyloxy)-2-
propoxycarbonyloxyrnethyl) ester,
phosphonoformic acid, mono-(4-N-valyloxy)cyclohexanoyloxymethyl) ester
to phosphonoformic acid, mono-(1-valyloxy-2-methylpropane-2-
aminocarbonyloxymethyl) ester
phosphonoformic acid, mono-(1-(2-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
carbonyloxymethyl) ester
phosphonoformic acid, mono-((1,3-di-(valyloxy)propyl-2-oxycarbonyioxy methyl)
15 ester
phosphonoformic acid, mono-(2-hydroxymethyl-2-rnethylpropionyloxymethyl) ester
phosphonoformic acid, mono-(2-(L-valyloxymethyl)-2-ethyl butyroyloxymethyl)
ester
phosphonoformic acid, mono-(3-(L-valyloxy)-2-methyl-propionyloxymethyl) ester
2o phosphonoformic acid, mono-(3-(L-valyloxy)-2-methyl, 2-hydroxymethyl-
propionyloxymethyl) ester
the corresponding bis esters, the corresponding compounds additionally bearing
a
conventional carboxyl ester, and pharmaceutically acceptable salts thereof.
25 An especially favoured group comprises:
phosponoformic acid di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl)
ester
phosponoformic acid mono-(2-methyl-2-(D-valyloxyrnethyl) propionyloxymethyl)
ester
phosponoformic acid di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
30 phosponoformic acid mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl)
ester,
phosponoformic acid di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
phosponoformic acid mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester
and
especially
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phosponoformic acid di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl)
ester
phosponoformic acid mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl)
ester;
and pharmaceutically acceptable salts and conventional pharmaceutical carboxy-
esters thereof.
A further useful group of compounds comprises:
phosponofonnic acid mono-(2-methyl-2-(L-isoleucyloxymethyl)
propionyloxymethyl) ester,
to phosponoformic acid mono-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyI)
ester,
phosponoformic acid mono-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl)
ester,
phosponoformic acid mono-{2-methyl-2-(L-alanyloxymethyl) propionyloxyrnethyl)
15 ester,
phosponofonmic acid mono-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl)
ester,
and the corresponding bis esters, conventional pharmaceutically acceptable
carboxy
esters and pharmaceutically acceptable salts thereof.
A further group of useful compounds comprises:
phosponoformic acid mono-L-valyloxymethyl ester,
phosponoformic acid di-L-valyloxymethyl ester,
phosponoformic acid mono-L-leucyloxymethyl ester,
2s phosponoformic acid di-L-leucyloxymethyl ester,
phosponoformic acid mono-L-isoleucyloxymethyl ester,
phosponoformic acid di-L-isoleucyloxymethyl ester,
phosponoformic acid mono-L-t-leucyloxymethyl ester,
phosponoformic acid di-L-t-leucyloxymethyl ester,
3o phosponoformic acid mono-L-alanyloxymethyl ester,
phosponoformic acid di-L-alanyloxymethyl ester,
conventional pharmaceutically acceptable carboxy esters and pharmaceutically
acceptable salts thereof.
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16
This aspect of the invention further provides a method for the treatment or
prophylaxis of virus infections comprising the oral administration of an
effective
amount of a compound of the invention to a mammal (including humans) in need
thereof. Viral infections include herpesvirus infections such as HSV-1, HSV-2,
VZV,
CMV, HHV6, HHV8 and retroviruses such as HIV-1 and HIV-2. The invention
further provides the use of the compounds defined above in medicine and the
use of
these compounds in the preparation of a medicament for the prophylaxis or
treatment
of viral infections.
A further class of phosphonates which are amenable to the invention and which
share
a structural similarity with phosphonoformic acid are the (3-
phosphonocarboxylic
acid farnesyl protein transferase inhibitors, especially those of the of the
formula
PF4:
IS
R4~ O RaR O O O- Rf2
RZ.-O- ()qi ()qr-T~1I--O O-P N~
Rf3
R4~ RaR ~ O
Rf1
or
O O- Rf2
R4R
R2 - O - Uqi-' Ring ~)Gr -T ~11- O
Rf3
R4R, O O
Rf1
where RF 1 is H or a further structure of formula IIa or IIb
Rf2 is H or a conventional pharmaceutically acceptable ester,
Rf3 is a polyunsaturated, branched C~zz alkyl,
RZ is the acyl residue of an aliphatic amino acid,
CA 02325523 2000-09-21
R4Land RdL' are independently H, C1_3 alkyl, hydroxymethyl, C3-6cycloalkyl, C1-
3alkyl-ClC6cycloalkyl phenyl or benzyl,
R4R and R4R' are independently H, CI_3 alkyl or phenyl
ql is 0-3, qr is 0-3,
T is a bond, -NR4- or -O-
R4 is H or C1_3alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle
Currently favoured values in Formula PF4 include: R4R and R4R' are preferably
H
and/or R4L and R4L' are preferably ethyl or especially methyl. T is preferably
-O- or
more preferably a bond. Preferably qr is 1 or more preferably 0 and ql is 0 or
preferably 1. If Rf 1 is a further ester it is convenient if it is identical
to other linker-R2
moiety. Conventional pharmaceutically acceptable esters for Rf2 include the
methyl,
ethyl and isopropyl esters or alternatively a similar structure to the linker-
R2 moiety as
envisaged in our copending PCT/SE99/00194 (W099 41275). A convenient
polyunsaturated alkyl Rf3 has the formula:
a ~~ U
where the asterisk shows the point of attachment.
Other structurally similar phosponates include a-phosphonosulphonates such as
squalene synthase inhibitors of the formula PFS:
~M~NDED SH~~T
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18
R aR O'~o' O Rf2
4L ~ R
R2-O- ()qi ()qr-T~O O-
, Rf3
O
R4~ R4R
Rf1
or
O
R O: S-O-Rf2
4R
R2 - O - ()qi- Ring - ()4r -T ~ O -
Rf3
O
R4R
Rf1
where RF1 is H or a further structure of formula IIa or IIb
Rf2 is H or a conventional pharmaceutically acceptable ester a further
structure of formula IIa or IIb
Rf3 is a polyunsaturated, branched C6_zz alkyl,
Rz is the acyl residue of an aliphatic amino acid,
R4Land R4~' are independently H, C,_, alkyl, hydroxymethyl, C,-bcycloalkyl, C,-
3alkyl-C,Cbcycloalkyl phenyl or benzyl,
to R4R and R4R' are independently H, C,_3 alkyl or phenyl
ql is 0-3, qr is 0-3,
T is a bond, -NR4- or -O-
R4 is H or C,_3alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle.
1s
Currently favoured values in Formula PFS include: R4R and R4R' are preferably
H
and/or R4L and R4L' are preferably ethyl or especially methyl. T is preferably
-O- or
more preferably a bond. Preferably qr is 1 or more preferably 0 and ql is 0 or
preferably 1. If Rfl is a further ester it is convenient if it is identical to
other linker-
2o Rz moiety. Conventional pharmaceutically acceptable esters for Rf2 include
the
methyl, ethyl and isopropyl esters, or alternatively a similar structure to
the linker-Rz
moiety. A convenient polyunsaturated alkyl Rf3 has the formula:
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19
where the asterisk denotes the point of attachment.
The aspects of the invention immediately above further provide respective
methods
for the treatment or prophylaxis of neoplasma or cholesterol disorders
comprising the
oral administration of an effective amount of a compound of the invention to a
mammal (including humans) in need thereof. The invention further provides the
use
of the compounds defined above in medicine and the use of these compounds in
the
1 o preparation of a medicament for the prophylaxis or treatment of neoplasma
or
cholesterol disorders.
A particularly preferred group of phosphorous containing drugs are the
bisphosphonates active in bone and calcium metabolism. Favoured bis-
phosphonates
15 drugs have the formula:
O,, O
HO-P P~-OH
HO OH
X
where
X is H, halo, hydroxy; and Y is
a) C,_,o alkyl, optionally substituted with
2o heterocycle,
-NRaRe, where Ra and Rb are independently
hydrogen, C,_6 alkyl or join together to form a 5 to 7
membered ring, optionally containing a further hetero
atom,
25 OH, halo, -S(C,_6 alkyl), phenyl, -C,_, cycloalkyl, (optionally substituted
with -NRaRb or OH);
b) C3_, cycloalkyl, optionally substituted with
-1VK,Rb, OH, halo, -S(C,_6 alkyl), phenyl, morpholino or pyridyl;
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c) halo;
d) piperidinyl;
e) pyrrolidinyl;
f) -S(C,_6 alkyl), optionally substitued with
5 -NRaRb, OH, halo or phenyl;
g) -S-phenyl, optionally substituted with
halo, nitro, C,_6 alkyl, C,_6 alkoxy, trifluormethyl, -CONRaRe or
-COOH.
1o Preferred bis-phosphonates include alendronate (X is hydroxy, Y is
NHZCHZCHZCHZ-), clodronate (X is chloro, Y is chloro), etidronate (X is
hydroxy, Y
is CH3-), pamidronate (X is hydroxy, Y is NH~CHZCH,-), ibandronate (X is
hydroxy,
Y is N (CHzCH2CHZCH2CH3)(CH3)CHzCH2-), tiludronate (X is H, Y is
4-chlorophenylthio-), risedronate (X is hydroxy, Y is 3-pyridinylmethylene-)
and
15 zoledronate (X is hydroxy, Y is (2-(1H-imidazol-1-yl)methylene-)
Preferred compounds within this bis-phosphonate aspect of the invention thus
include those of the formula A1:
O YY O
Ra1-O-P P-O-Ra2
~O ~ ~O\
Ra3 Ra4
2o A-1
wherein YY and XX have the following values:
NHZ(CHZ)3- OH (alendronate)
NHZ(CHZ)Z- OH (pamidronate)
cycloheptylamino- H (cimadronate)
chloro- chloro (clodronate)
pyrrolidin-1-ylCH2CH2- OH ~ (EB 1053)
CH3- OH (etidronate)
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21
methylpentylaminoCHZCH2- OH (ibandronate)
dimethylaminoCHZCHZ OH (olpadronate)
pyridin-3-ylCH2- OH (risedronate)
(4-chlorophenyl)-thio- H (tiludronate)
imidazo-(1,2-a)pyridin-3-ylCHz-OH (YH 529)
1 H-imidazol-1 ylCHz- OH (zoledronate)
wherein amino groups on YY can be optionally substituted with conventional
pharmaceutically acceptable amide groups such as -C(=O)C,_6alkyl or an
aminoacyl
or peptidyl derivative, as described in WO 96/31227;
and wherein at least one of Ral-Ra4 is a structure of the formula
R4L O R4R
R2-_ O - ()qi ()qr-T --~ o
R4L~ RaR
or
R4R
R2-O- ()ql- Ring ()qr-T --~O
R4R
where
RZ is the acyl residue of an aliphatic amino acid,
R4Land R4L' are independently H, C,_, alkyl, hydroxymethyl, C3-6cycloalkyl, C,-
3alkyl-C,Cbcycloalkyl, phenyl or benzyl,
R4R and R4R' are independently H or C,_3 alkyl
ql is 0-3, qr is 0-3,
2o T is a bond, -NR4- or -O
R4 is H or C,_,alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
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22
and the remainder of Ral-4 are hydrogen or conventional pharmaceutically
acceptable esters. Preferably Ral and Ra2 comprise the same structure of
formula IIa
or IIb and Ra3 and Ra4 are H. A free amine group on YY could also be protected
with the same linker (RZ')k-Rz structure, as envisaged in our copending
application no
PCT/SE99/00194.
In formula A-1, R4R and R4R' are preferably H and/or R4L and R4L' are
preferably
ethyl or especially methyl. T is preferably -O- or more preferably a bond.
Preferably
qr is 1 or more preferably 0 and ql is 0 or more preferably 1.
Representative compounds within formula A-1 thus include;
(4-amino-1-hydroxybutylidine)-bisphosphonate, di(2-methyl-2-(L-valyloxymethyl)
propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di(2-methyl-2-(L-valyloxy)
propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di (2-(L-valyloxy)-3-methyl-(S)-
(+)-
butyryloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di (2-(-L-valyloxy)-2-phenyl-DL-
acetyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di ((1,3-di-valyloxy)propyl-2-
oxycarbonyloxy methyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di (2-L-valyloxy)-DL-
propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(5-(L-valyloxy)-2,2-
2s dimethylvaleryloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-((2-(L-valyloxy)-
ethoxycarbonyloxy) methyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, bis [4-(L-valyloxy)-
butanoyloxymethyl] ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(4-(L-valyloxy)
benzoyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(3-(3,4-di-(L-valyloxy)
phenyl)
propionyloxymethyl) ester,
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23
(4-amino-I-hydroxybutylidine)-bisphosphonate, di- (2-methyl-1-(L-valyloxy)-2-
propoxycarbonyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(4-
valyloxy)cyclohexanoyloxymethyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-[N-(2-L-valyloxy-1,1-dimethyl-
ethyl) aminocarbonyloxymethylJ ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, di- ( I -(2-L-valyloxyethyl)-6-
oxo-
1,6-dihydro-pyridine-3-carbonyloxymethyl)
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-((1,3-di-(valyloxy)propyl-2-
to oxycarbonyloxy methyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(2-hydroxymethyl-2-
methylpropionyloxymethyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, di-(2-(L-valyloxymethyl)-2-ethyl
15 butyroyloxymethyl) ester
(4-amino-I-hydroxybutylidine)-bisphosphonate, di (3-(L-valyloxy)-2-methyl-
propionyloxymethyl) ester
(4-amino-I-hydroxybutylidine)-bisphosphonate, di (3-(L-valyloxy)-2-methyl,2-
hydroxymethyl-propionyloxymethyl) ester
2o 1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di(2-methyl-2-
(L-
valyloxyrnethyl) propionyloxymethyl) ester,
1-hydroxy-2-{IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di(2-methyl-2-(L-
valyloxy) propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di (2-(L-valyloxy)-
3-
25 methyl-(S)-(+)-butyryloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di (2-(-L-
valyloxy)-2-
phenyl-DL-acetyloxymethyl) ester,
I-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di ((1,3-di-
valyloxy)propyl-2-oxycarbonyloxy methyl) ester,
30 1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di (2-L-
valyloxy)-DL-
propionyloxymethyl) ester,
1-hydroxy-2-( 1 H-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(5-(L-
valyloxy)-
2,2-dimethylvaleryloxymethyl) ester,
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24
1-hydroxy-2-(IH-imidazolyl-I-yl)ethylidene-bis phosphonate, di-((2-(L-
valyloxy}-
ethoxycarbonyloxy) methyl) ester,
1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, bis [4-(L-
valyloxy)-
butanoyloxymethyl] ester,
I-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(4-(L-valyloxy)
benzoyloxymethyl) ester,
1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(3-(3,4-di-(L-
valyloxy) phenyl) propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di- (2-methyl-1-(L-
1o valyloxy)-2-propoxycarbonyloxymethyl) ester,
I-hydroxy-2-(1H-imidazolyl-I-yl)ethylidene-bis phosphonate, di-(4-
valyloxy)cyclohexanoyloxymethyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di-[N-(2-L-
valyloxy-
1,1-dimethyl-ethyl) aminocarbonyloxymethyl]ester
15 1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di- (1-(2-L-
valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester,
1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di-((1,3-di-
(valyloxy)propyl-2-oxycarbonyloxy methyl) ester
I-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(2-
hydroxymethyl-
20 2-methylpropionyloxymethyl) ester
1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(2-(L-
valyloxymethyl)-2-ethyl butyroyloxymethyl) ester
1-hydroxy-2-(IH-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(3-(L-valyloxy)-
2-
methyl-propionyloxymethyl) ester
25 1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, di-(3-(L-
valyloxy}-2-
methyl,2-hydroxymethyl-propionyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate di(2-methyl-2-(L-
valyloxyrnethyl) propionyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di(2-methyl-2-(L-valyloxy)
3o propionyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di (2-(L-valyloxy)-3-
methyl-
(S)-(+)-butyryloxymethyl) ester,
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I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di (2-(-L-valyloxy)-2-
phenyl-
DL-acetyloxymethyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di ((1,3-di-
valyloxy)propyl-2-
oxycarbonyloxy methyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di (2-L-valyloxy)-DL-
propionyloxymethyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(5-(L-valyloxy)-2,2-
dimethylvaleryloxymethyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-((2-(L-valyloxy)-
10 ethoxycarbonyIoxy) methyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, bis [4-(L-valyloxy)-
butanoyloxymethyl] ester,
I-hydroxy-2-(pyrid-3-yI)ethylidene bis-phosphonate, di-(4-(L-valyloxy)
benzoyloxymethyl) ester,
15 I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(3-(3,4-di-(L-
valyloxy)
phenyl) propionyloxymethyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di- (2-methyl-1-(L-
valyloxy)-2-
propoxycarbonyloxymethyl) ester,
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(4-
20 valyloxy)cyclohexanoyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-[N-(2-L-valyloxy-1,1-
dimethyl-ethyl) aminocarbonyloxymethyl] ester
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di- (I-(2-L-valyloxyethyl)-
6-
oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester
25 1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-((1,3-di-
(valyloxy)propyl-2-
oxycarbonyloxy methyl) ester
I-hydroxy-2-(pyrid-3-y1)ethylidene bis-phosphonate, di-(2-hydroxymethyl-2-
methylpropionyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(2-(L-valyloxymethyl)-2-
ethyl butyroyloxymethyl) ester
I-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(3-(L-valyloxy)-2-
methyl-
propionyloxymethyl) ester
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1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, di-(3-(L-valyloxy)-2-
methyl, 2-
hydroxymethylpropionyloxymethyl) ester
and pharmaceutically acceptable salts thereof.
A further group of representative compounds include:
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono(2-methyl-2-(L-
valyloxymethyl) propionyloxyrnethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono(2-methyl-2-(L-valyloxy)
propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono(2-(L-valyloxy)-3-methyl-(S)-
(+)-butyryloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono(2-(-L-valyloxy)-2-phenyl-
DL-acetyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono((1,3-di-valyloxy)propyl-2-
oxycarbonyloxy methyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono(2-L-valyloxy)-DL-
propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-(5-(L-valyloxy)-2,2-
dimethylvaleryloxymethyl) ester,
(4-amino-i-hydroxybutylidine)-bisphosphonate, mono-((2-(L-valyloxy)-
ethoxycarbonyloxy) methyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono [4-( L-valyloxy)-
butanoyloxymethyl] ester,
(4-amino-l-hydroxybutylidine)-bisphosphonate, mono-(4-(L-valyloxy)
benzoyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-(3-(3,4-di-(L-valyloxy)
phenyl) propionyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono- (2-methyl-1-(~.-valyloxy)-
2-
propoxycarbonyloxymethyl) ester,
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-{4-
valyloxy)cyclohexanoyloxymethyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-[N-(2-L-valyloxy-1,1-
dimethyl-ethyl) aminocarbonyloxymethyl] ester
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27
(4-amino-I-hydroxybutylidine)-bisphosphonate, mono-(1-(2-L-valyloxyethyl)-6-
oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester
(4-amino-I-hydroxybutylidine)-bisphosphonate, mono-((1,3-di-(valyloxy)propyl-2-
oxycarbonyloxy methyl) ester
(4-amino-I-hydroxybutylidine)-bisphosphonate, mono-(2-hydroxymethyl-2-
methylpropionyloxymethyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-(2-(L-valyloxymethyl)-2-
ethyl butyroyloxymethyl) ester
(4-amino-1-hydroxybutylidine)-bisphosphonate, mono-(3-(L-valyloxy}-2-methyl-
l0 propionyloxymethyl) ester
(4-amino-I-hydroxybutylidine)-bisphosphonate, mono-(3-(L-valyloxy}-2-methyl,2-
hydroxymethylpropionyloxymethyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono(2-methyl-2-(L-
15 valyloxymethyl) propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono(2-methyl-2-(L-
valyloxy) propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono (2-(L-
valyloxy)-3-methyl-(S)-(+)-butyryloxymethyl) ester,
2o 1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono (2-(-L-
valyloxy)-2-phenyl-DL-acetyloxymethyl) ester,
I-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono ((1,3-di-
valyloxy}propyl-2-oxycarbonyloxy methyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono (2-L-
valyloxy)-
2s DL-propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(S-(L-
valyloxy)-2,2-dimethylvaleryloxymethyl) ester,
1-hydroxy-2-(IH-imidazolyl-I-yl)ethylidene-bis phosphonate, mono- ((2-(L-
valyloxy)-ethoxycarbonyloxy) methyl) ester,
30 1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono [4-(L-
valyloxy)-butanoyloxymethyl] ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(4-(L-
valyloxy)
benzoyloxymethyl) ester,
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1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(3-(3,4-di-(L-
valyloxy) phenyl) propionyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono- (2-methyl-1-
(L-valyloxy)-2-propoxycarbonylmethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(4-N-
valyloxy)cyclohexanoyloxymethyl) ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-[N-(2-L-
valyloxy-1,1-dimethyl-ethyl) aminocarbonyloxymethyl] ester,
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(1-(2-L-
valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-((1,3-di-
(valyloxy)propyl-2-oxycarbonyloxy methyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(2-
hydroxymethyl-2-methylpropionyloxymethyl) ester
~5 1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(2-(L-
valyloxymethyl)-2-ethyl butyroyloxymethyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(3-(L-
valyloxy)-2-methyl-propionyloxymethyl) ester
1-hydroxy-2-(1H-imidazolyl-1-yl)ethylidene-bis phosphonate, mono-(3-(L-
20 valyloxy)-2-methyl,2-hydroxymethyl-propionyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate mono(2-methyl-2-(L-
valyloxymethyl) propionyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono(2-methyl-2-(L-
valyloxy)
propionyloxymethyl) ester,
25 1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono (2-(L-valyloxy)-3-
methyl-(S)-(+)-butyryloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono (2-(-L-valyloxy)-2-
phenyl-DL-acetyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono((1,3-di-
valyloxy)propyl-
3o 2-oxycarbonyloxy methyl) ester,
1-hydroxy-2-(pyrid-3-yI)ethylidene bis-phosphonate, mono(2-L-valyloxy)-DL-
propionyloxymethyl) ester,
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1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(5-(L-valyloxy)-2,2-
dimethylvaleryloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-((2-(L-valyloxy)-
ethoxycarbonyloxy) methyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono [4-( L-valyloxy)-
butanoyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(4-(L-valyloxy)
benzoyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(3-(3,4-di-(L-
valyloxy)
1o phenyl) propionyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono- (2-methyl-1-(L-
valyloxy)-2-propoxycarbonyloxymethyl) ester,
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(4 -
valyloxy)cyclohexanoyloxymethyl) ester
15 1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-[N-(2-L-valyloxy-
1,1-
dimethyl-ethyl) aminocarbonyloxymethyl] ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(1-(2-L-
valyloxyethyl)-
6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-((1,3-di-
20 (valyloxy)propyl-2-oxycarbonyloxy methyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(2-hydroxymethyl-2-
methylpropionyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(2-(L-valyloxymethyl)-
2-ethyl butyroyloxymethyl) ester
25 1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(3-(L-valyloxy)-2-
methyl-propionyloxymethyl) ester
1-hydroxy-2-(pyrid-3-yl)ethylidene bis-phosphonate, mono-(3-(L-valyloxy)-2-
methyl, 2-hydroxymethyl-propionyloxymethyl) ester
and pharmaceutically acceptable salts thereof.
A favoured group of compounds within formula A-1 include:
alendronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
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alendronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
alendronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
alendronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
alendronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
5 alendronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
alendronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
risedronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
risedronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
10 risedronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
risedronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
risedronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
zoledronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
~s zoledronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
zoledronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
zoledronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
2o zoledronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
pamidronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
pamidronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-valyloxy} propionyloxymethyl) ester,
25 pamidronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
pamidronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
cimadronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
3o cimadronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
cimadronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
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cimadronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
clodronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
clodronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
clodronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
clodronate mono-(2-methyl-2-(D-valyloxy) propionyloxyrnethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate mono-(2-methyl-2-(L-
1o valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-(D-
valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate mono-(2-methyl-2-(D-
valyloxymethyl) propionyloxymethyl) ester,
15 [1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-
(L-
valyloxy) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate mono-(2-methyl-2-(L-
valyloxy) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-(D-
20 valyloxy) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate mono-(2-methyl-2-(D-
valyloxy) propionyloxymethyl) ester,
etidronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
25 etidronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
etidronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
etidronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
30 olpadronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
olpadronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
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olpadronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
olpadronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(D-valyloxy) propionyIoxymethyl) ester,
olpadronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
ibandronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
ibandronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
ibandronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
1o ibandronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
ibandronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate mono-(2-
methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(D-valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine)-bis-phosphonate mono-(2-
methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-valyloxy) propionyloxymethyl) ester,
2o [1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate mono-(2-
methyl-2-(L-valyloxy) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(D-valyloxy) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate mono-(2-
methyl-2-(D-valyloxy) propionyloxymethyl) ester,
tiludronate mono-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
tiludronate di-(2-methyl-2-{D-valyloxymethyl) propionyloxymethyl) ester,
tiludronate mono-(2-methyl-2-(D-valyloxymethyl) propionyloxymethyl) ester,
tiludronate di-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
3o tiludronate mono-(2-methyl-2-(L-valyloxy) propionyloxymethyl) ester,
tiludronate di-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
tiludronate mono-(2-methyl-2-(D-valyloxy) propionyloxymethyl) ester,
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and pharmaceutically acceptable salts thereof.
A particularly favoured group of compounds of the invention comprises:
alendronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-{L-valyloxymethyl) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
to [1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-
(L-
valyloxymethyl) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(L-valyloxyrnethyl) propionyloxymethyl) ester,
15 tiludronate di-(2-methyl-2-(L-valyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-valyloxymethyl) propionyloxymethyl) ester,
and pharmaceutically acceptable salts thereof, especially the alendronate,
zoledronate
and risedronate derivatives.
A further useful group of compounds within formula A-1 include:
alendronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
alendronate di-(2-methyl-2-(L-leucyloxymethyl) propionyloxymethyl) ester,
alendronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
alendronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
alendronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
risedronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
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zoledronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
zoledronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
parnidronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
pamidronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
~o cimadronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl)
ester,
cimadronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxyrnethyl) ester,
cimadronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
cimadronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
~ 5 clodronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl)
ester,
clodronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyioxymethyl) ester,
clodronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
clodronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
20 [1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-
(L-
isoleucyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrroiidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-(L-
leucyoxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-(L-t-
2s leucyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-{2-methyl-2-(L-
alanyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-3-(1-pyrrolidinyl)-propylidine]-bis-phosphonate di-(2-methyl-2-(L-
glycyloxymethyl) propionyloxymethyl) ester,
3o etidronate di-(2-methyl-2-(L-isoleucyloxyrnethyl) propionyloxymethyl)
ester,
etidronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
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etidronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
etidronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
s olpadronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
olpadronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(L-isoleucyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(L-leucyoxymethyl) propionyloxymethyl) ester,
I o ibandronate di-(2-methyl-2-(L-t-leucyloxymethyl) propionyloxymethyl)
ester,
ibandronate di-(2-methyl-2-(L-alanyloxymethyl) propionyloxymethyl) ester,
ibandronate di-(2-methyl-2-(L-glycyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-isoleucyloxymethyl) propionyloxymethyl) ester,
15 [1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-leucyoxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-t-leucyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
20 (L-alanyloxymethyl) propionyloxymethyl) ester,
[1-hydroxy-2imidazo-(1,2-a)pyridin-3-ylethylidine]-bis-phosphonate di-(2-
methyl-2-
(L-glycyloxymethyl) propionyloxymethyl) ester,
and pharmaceutically acceptable salts thereof, especially the alendronate,
zoledronate
and risedronate derivatives.
A further group of representative compounds comprises
alendronate mono-L-valyloxymethyl ester,
alendronate di-L-valyloxymethyl ester,
alendronate mono-L-leucyloxymethyl ester,
3o alendronate di-L-leucyloxymethyl ester,
alendronate mono-L-isoleucyloxymethyl ester,
alendronate di-L-isoleucyloxymethyl ester,
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alendronate mono-L-t-leucyloxymethyl ester,
alendronate di-L-t-leucyloxymethyl ester,
alendronate mono-L-alanyloxymethyl ester,
alendronate di-L-alanyloxymethyl ester,
risedronate mono-L-valyloxyrnethyl ester,
risedronate di-L-valyloxymethyl ester,
risedronate mono-L-leucyloxymethyl ester,
risedronate di-L-leucyloxymethyl ester,
risedronate mono-L-isoleucyloxymethyl ester,
1 o risedronate di-L-isoleucyloxymethyl ester,
risedronate mono-L-t-leucyloxymethyl ester,
risedronate di-L-t-leucyloxymethyl ester,
risedronate mono-L-alanyloxymethyl ester,
risedronate di-L-alanyloxymethyl ester,
zoledronate mono-L-valyloxymethyl ester,
zoledronate di-L-valyloxymethyl ester,
zoledronate mono-L-leucyloxymethyl ester,
zoledronate di-L-leucyloxymethyl ester,
zoledronate mono-L-isoleucyloxymethyl ester,
2o zoledronate di-L-isoleucyloxymethyl ester,
zoledronate mono-L-t-leucyloxymethyl ester,
zoledronate di-L-t-leucyloxymethyi ester,
zoledronate mono-L-alanyloxymethyl ester,
zoledronate di-L-alanyloxymethyl ester,
and pharmaceutically acceptable salts thereof.
This aspect of the invention further provides a method for the treatment or
prophylaxis of bone and calcium disorders comprising the oral administration
of an
effective amount of a compound of the invention to a mammal (including humans)
in
3o need thereof. Bone and calcium disorders include osteoporosis, Paget's
disease,
hypercalcaemia of malignancy, tooth loss, bone loss in immunotherapy and
rheumatoid arthritis, decreasing fracture, post orthopedic prosthesis and
inhibiting
ossification. The invention further provides the use of the compounds defined
above
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37
in medicine and the use of these compounds in the preparation of a medicament
for
the prophylaxis or treatment of bone and calcium disorders.
A still further preferred group of prodrugs of the invention are those based
on
fosinoprilate having the formula PF3:
O
\ O H OH
~~~N
j~P
O O
R4R ~ R4R'
O - ()qi ~)qr-T -~- O
O
R4L'
or / H H
N
P
O O
R3R ~ R3R'
R2 -O_ ()qi-ring- OGr-T-~-O
O
where
RZ is the acyl residue of an aliphatic amino acid,
1o R4Land R4L' are independently H, C,_, alkyl, hydroxymethyl, C3-bcycloalkyl,
C,-
3alkyl-C,C6cycloalkyl phenyl or benzyl,
R4R and R4R' are independently H or C,_3 alkyl
ql is 0-3, qr is 0-3,
T is a bond, -NR3- or -O-
15 R3 is H or C,_3alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
and pharmaceutically acceptable salts thereof.
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38
In formula PF3, R4R and R4R' are preferably H and/or R4L and RqL' are
preferably
ethyl or especially methyl. T is preferably -O- or more preferably a bond.
Preferably
qr is 1 or more preferably 0 and ql is 0 or preferably 1.
Representative compounds within formula PF3 thus include
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (2-
methyl-2-
(L-valyloxymethyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinylJacetyl]-L-proline, (2-
methyl-2-
io (L-valyloxy) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (2-(L-
valyloxy)-3-methyl-(S)-(+)-butyryloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (2-(-L-
valyloxy)-2-phenyl-DL-acetyloxymethyl) ester,
15 (4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinylJacetyl]-L-proline, ((1,3-
di-
valyloxy)propyl-2-oxycarbonyloxy methyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (2-L-
valyloxy)-DL-propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (5-(L-
2o valyloxy)-2,2-dimethylvaleryloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, ((2-(L-
valyloxy)-ethoxycarbonyloxy) methyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, [4-( L-
valyloxy)-butanoyloxymethylJ ester,
25 (4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (4-(L-
valyloxy) benzoyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (3-(3,4-
di-(L-
valyloxy) phenyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, (2-
methyl-1-
30 (L-valyloxy)-2-propoxycarbonyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline,(4-N-
valyloxy)cyclohexanoyloxymethyl) ester
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39
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline, [N-(2-L-
valyloxy-1,1-dimethyl-ethyl) aminocarbonyloxymethyl] ester
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline,( 1-(2-L-
valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester
s (4S)-4-cyclohexyl-1-[[(R}(4-phenylbutyl)phosphinyl]acetyl]-L-proline, ((1,3-
di-
(valyloxy)propyl-2-oxycarbonyloxy methyl) ester
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline(2-
hydroxymethyl-2-methylpropionyloxymethyl) ester
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline(2-(L-
1 o valyloxymethyl)-2-ethyl butyroyloxymethyl) ester
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (3-(L-
valyloxy)-2-methyl-propionyloxymethyl) ester
4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (3-(L-
valyloxy)-2-methyl,2-hydroxymethyl-propionyloxymethyl) ester
1 s and pharmaceutically acceptable salts thereof.
A favoured group of compounds within formula PF3 comprises:
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
(L-valyloxymethyl) propionyloxymethyl) ester,
20 (4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-
methyl-2-
(D-valyloxymethyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[((R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
(L-valyloxy) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[((R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline mono-(2-
2s methyl-2-(D-valyloxy) propionyloxymethyl) ester,
and especially;
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
(L-valyloxyrnethyl) propionyloxymethyl) ester:
and pharmaceutically acceptable salts thereof
A further group of convenient compounds comprises
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(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetylJ-L-proline (2-methyl-
2-
(L-isoleucyloxymethyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
(L-leucyoxymethyl) propionyloxymethyl) ester,
5 (4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-
methyl-2-
(L-t-leucyloxymethyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
(L-alanyloxymethyl) propionyloxymethyl) ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline (2-methyl-
2-
10 (L-glycyloxymethyl) propionyloxymethyl) ester,
and pharmaceutically acceptable salts thereof.
A further group of useful compounds comprises:
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline L-
15 valyloxymethyl ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline L-
leucyloxymethyl ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline L-
isoleucyloxymethyl ester,
2o (4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline L-t-
leucyloxymethyl ester,
(4S)-4-cyclohexyl-1-[[(R)(4-phenylbutyl)phosphinyl]acetyl]-L-proline L-
alanyloxymethyl ester,
and pharmaceutically acceptable salts thereof.
This aspect of the invention further provides a method for the treatment or
prophylaxis of hypertension comprising the oral administration of an effective
amount of a compound defined above to a mammal (including humans) in need
3o thereof. The invention further provides the use of these compounds in
medicine and
their use in the preparation of a medicament for the treatment or prophylaxis
of
hypertension
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41
A further phosphonate compound amenable to the prodrugs of the invention are
the
neutral endopeptidase inhibitors such as CGS-24592 (Novartis), preferably
those of
the formula PF6:
I
i
I
R4L O R4R O ~ Ri2
R2-O- Oqi Oqr -T -~--O p- ~p~H NH~O
R4L' R4R' ~ O O
Rf1
w
or I
I~1~"
Rf2
O R4R O
R2-O- ()ql- Ring - ()qr -T ~-O _ ~p~H NH~O
R4R' ~ O O
Rf1
where RF1 is H or a further structure of formula IIa or IIb
1o RfZ is H or a conventional pharmaceutically acceptable ester,
Rz is the acyl residue of an aliphatic amino acid,
R4Land R4L' are independently H, C,_3 alkyl, hydroxymethyl, C3-bcycloalkyl, C,-
3alkyl-C,C6cycloalkyl phenyl or benzyl,
R4R and R4R' are independently H or C,_3 alkyl
ql is 0-3, qr is 0-3,
T is a bond, -NR4- or -O-
R4 is H or C,_,alkyl;
ring is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
2o and pharmaceutically acceptable salts thereof.
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42
Currently favoured values in Formula PF6 include: R4R and R4R' are preferably
H
and/or R4L and R4L' are preferably ethyl or especially methyl. T is preferably
-O- or
more preferably a bond. Preferably qr is 1 or more preferably 0 and ql is 0 or
preferably 1. If Rfl is a further ester it is convenient if it is identical to
other linker-
RZ moiety. Conventional pharmaceutically acceptable esters for Rf2 include the
methyl, ethyl and isopropyl esters.
Compounds of the invention are typically prepared by esterifying the mother
1 o compound in which exposed functions such as the 4-amino group of
alendronate or
the carboxy group of foscarnet are protected with conventional amine and
carboxy
etc protecting groups respectively, with a structure of the formula IIa or
IIb, the
preparation of which are described in the abovementioned PCT/SE99/00194:
R4~ O R4R
*R2 -O- ()ql ()qr-T ---~-O halo
R4~ R4R
I I a'
or
R4R
*R2 O- Oql Ring - ()qr-T --~1- O halo
R4R
~s Ilb'
*RZ is the acyl residue of an aliphatic amino acid, N-protected with a
conventional N-
protecting group such as CBz, Fmoc or Boc
R4Land R4L' are independently H, C,_3 alkyl, hydroxymethyl, C3-bcycloalkyl, C,-
3alkyl-C,Cbcycloalkyl phenyl or benzyl,
2o R4R and R4R' are independently H or C,_3 alkyl
ql is 0-3, qr is 0-3,
T is a bond, -NR3- or -O-
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WO 99/51613 PCT/SE99/00528
R3 is H or C,_3alkyl;
43
"ring" is an optionally substituted aromatic or non-aromatic, hetero-or
carbocycle;
and halo is bromo, chloro and especially iodo. Alternatively, iodomethyl N-
protected
aminoacid esters are employed for linker-RZ structures of the formula:
R2~" O ~
Hydroxymethyl bearing linkers such as those within formula IIa are prepared by
methoxybenzylation of the free carboxy group of a bis hydroxy compound such as
glycerol or bis 2,2 hydroxymethyl propionic acid, CBz monoprotection of one of
the
hydroxy groups, esterification with the N-protected RZ group, debenzylation,
reaction
with chloroiodomethyl and iodination as shown below prior to esterificaiton to
a
phosphorous-containing pharmaceutical.
The mother compounds, that is the phosphorous containing drug, are all well
known
and readily accessible to those in the art.
In drugs comprising multiple phosphoryl/phosphonate/phosphinate functions, it
is
generally advantageous that an hydroxy group on each phosphorous moiety is
esterified with a linker (-RZ')k-Rz structure such as those of formula IIa or
IIb etc.
2o Regioselective protecting groups which bridge the phosphate groups of bis
phosphonates and thus assist mono and diacylation include Si compounds such as
dichlorotetraisopropyldisosiloxane or cresols.
Methodology for the derivatisation of phosphorous-containing compounds with
acyloxyalkyl groups and which can be used analogously for the coupling of the
-linker (-Rz')k-RZ structure is described in US patent 5 227 506, WO 94/13682,
WO
94/13324, WO 98/04569, Starret et al J Med Chem 37 1857-1864 (1994) and Iyer
et
al Tetrahedron Lett 30 7141-7144 (1989). A number of approaches are also
illustrated in the examples below. The N-protecting group on group RZ is
removed by
conventional methodology.
The term "N-protecting group" or "N-protected" as used herein refers to those
groups
intended to protect the N-terminus of an amino acid or peptide or to protect
an amino
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44
group against undesirable reactions during synthetic procedures. Commonly used
N-
protecting groups are disclosed in Greene, "Protective Groups in Organic
Synthesis"
(John Wiley & Sons, New York, 1981 ), which is hereby incorporated by
reference.
N-protecting groups include acyl groups such as formyl, acetyl, propionyl,
pivaloyl,
t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoracetyl, trichloroacetyl,
phthalyl,
o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-
bromobenzoyl,
4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl, p-
toluenesulfonyl, and the like, carbamate forming groups such as
benzyloxycarbonyl,
p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,
to p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-
bromobenzyloxycarbonyl,
3,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl,
2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl,
1-(p-biphenylyl)-1-methylethoxycarbonyl, a,a-dimethyl-3,5-
dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butoxycarbonyl,
15 diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl,
rnethoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl,
phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl,
cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl,
phenylthiocarbonyl, and the like; alkyl groups such as benzyl,
triphenylmethyl,
2o benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and
the like.
Favoured N-protecting groups include formyl, acetyl, allyl, F-moc, benzoyl,
pivaloyl,
t-butylacetyl, phenylsulfonyl, benzyl, t-butoxycarbonyl (BOC) and
benzyloxycarbonyl (Cbz).
25 Hydroxy and/or carboxy protecting groups are also extensively reviewed in
Greene
ibid and include ethers such as methyl, substituted methyl ethers such as
methoxymethyl, methylthiomethyl, benzyloxymethyl, t-butoxymethyl, 2-
methoxyethoxymethyl and the like, silyl ethers such as trimethylsilyl (TMS), t-
butyldimethylsilyl (TBDMS) tribenzylsilyl, triphenylsilyl, t-
butyldiphenylsilyl
3o triisopropyl silyl and the like, substituted ethyl ethers such as 1-
ethoxymethyl, 1-
methyl-1-methoxyethyl, t-butyl, allyl, benzyl, p-methoxybenzyl,
dipehenylmethyl,
triphenylmethyl and the like, aralkyl groups such as trityl, and pixyl (9-
hydroxy-9-
phenylxanthene derivatives, especially the chloride). Ester hydroxy protecting
groups
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WO 99/51613 PCT/SE99/00528
include esters such as formate, benzylformate, chloroacetate, methoxyacetate,
phenoxyacetate, pivaloate, adamantoate, mesitoate, benzoate and the like.
Carbonate
hydroxy protecting groups include methyl vinyl, allyl, cinnamyl, benzyl and
the like.
The compounds of the invention can form salts which form an additional aspect
of
the invention. Appropriate pharmaceutically acceptable salts of the compounds
of the
invention include salts of organic acids, especially carboxylic acids,
including but not
limited to acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate,
maleate,
malate, pantothenate, isethionate, adipate, alginate, aspartate, benzoate,
butyrate,
1o digluconate, cyclopentanate, glucoheptanate, glycerophosphate, oxalate,
heptanoate,
hexanoate, fumarate, nicotinate, palmoate, pectinate, 3-phenylpropionate,
picrate,
pivalate, proprionate, tartrate, lactobionate, pivolate, camphorate,
undecanoate and
succinate, organic sulphonic acids such as methanesulphonate,
ethanesulphonate,
2-hydroxyethane sulphonate, camphorsulphonate, 2-napthalenesulphonate,
1 s benzenesulphonate, p-chlorobenzenesulphonate and p-toluenesulphonate; and
inorganic acids such as hydrochloride, hydrobromide, hydroiodide, sulphate,
bisulphate, hemisulphate, thiocyanate, persulphate, phosphoric and sulphonic
acids.
The compounds of the invention may in some cases be isolated as the hydrate.
2o While it is possible for the active agent to be administered alone, it is
preferable to
present it as part of a pharmaceutical formulation. Such a formulation will
comprise
the above defined active agent together with one or more acceptable
carriers/excipients and optionally other therapeutic ingredients. The
carriers) must
be acceptable in the sense of being compatible with the other ingredients of
the
25 formulation and not deleterious to the recipient.
The formulations include those suitable for rectal, nasal, topical (including
buccal
and sublingual), vaginal or parenteral (including subcutaneous, intramuscular,
intravenous and intradermal) administration, but preferably the formulation is
an
30 orally administered formulation. The formulations may conveniently be
presented in
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46
unit dosage form, e.g. tablets and sustained release capsules, and may be
prepared by
any methods well known in the art of pharmacy.
Such methods include the step of bringing into association the above defined
active
agent with the carrier. In general, the formulations are prepared by uniformly
and
intimately bringing into association the active agent with liquid carriers or
finely
divided solid carriers or both, and then if necessary shaping the product. The
invention extends to methods for preparing a pharmaceutical composition
comprising
bringing a compound of the invention or its pharmaceutically acceptable salt
in
1o conjunction or association with a pharmaceutically acceptable carrier or
vehicle. If
the manufacture of pharmaceutical formulations involves intimate mixing of
pharmaceutical excipients and the active ingredient in salt form, then it is
often
preferred to use excipients which are non-basic in nature, i.e. either acidic
or neutral.
Formulations for oral administration in the present invention may be presented
as
discrete units such as capsules, cachets or tablets each containing a
predetermined
amount of the active agent; as a powder or granules; as a solution or a
suspension of
the active agent in an aqueous liquid or a non-aqueous liquid; or as an oil-in-
water
liquid emulsion or a water in oil liquid emulsion and as a bolus etc.
With regard to compositions for oral administration {e.g. tablets and
capsules), the
term suitable Garner includes vehicles such as common excipients e.g. binding
agents, for example syrup, acacia, gelatin, sorbitol, tragacanth,
polyvinylpyrrolidone
(Povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose,
hydroxypropylmethylcellulose, sucrose and starch; fillers and carriers, for
example
corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin,
mannitol,
dicalcium phosphate, sodium chloride and alginic acid; and lubricants such as
magnesium stearate, sodium stearate and other metallic stearates, glycerol
stearate
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47
stearic acid, silicone fluid, talc waxes, oils and colloidal silica.
Flavouring agents
such as peppermint, oil of wintergreen, cherry flavouring or the like can also
be used.
It may be desirable to add a colouring agent to make the dosage form readily
identifiable. Tablets may also be coated by methods well known in the art.
A tablet may be made by compression or moulding, optionally with one or more
accessory ingredients. Compressed tablets may be prepared by compressing in a
suitable machine the active agent in a free flowing form such as a powder or
granules, optionally mixed with a binder, lubricant, inert diluent,
preservative,
1o surface-active or dispersing agent. Moulded tablets may be made by moulding
in a
suitable machine a mixture of the powdered compound moistened with an inert
liquid
diluent. The tablets may be optionally be coated or scored and may be
formulated so
as to provide slow or controlled release of the active agent.
1s Other formulations suitable for oral administration include lozenges
comprising the
active agent in a flavoured base, usually sucrose and acacia or tragacanth;
pastilles
comprising the active agent in an inert base such as gelatin and glycerin, or
sucrose
and acacia; and mouthwashes comprising the active agent in a suitable liquid
Garner.
2o Detailed Description of the Invention
Various aspects of the invention will now be illustrated by way of example
only with
reference to the following non-limiting synthesis and biological examples:
25 Preparation of intermediates
Example P-I-1
N-BOC-L-isoleucine iodometh, l ester
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48
~ O
O- _N
H I
O
a) N-BOC-L-isoleucine chloromethyl ester
To a solution of N-BOC-L-isoleucine (23.1 g, 0.1 mol) in dioxane (500 mL), was
added dropwise a 40% aqueous solution of tetrabutylammonium hydroxide (65.6
mL, 0.1 mol). After stirring for 15 min, the solution was evaporated to
dryness
through co-evaporation with dioxane and toluene. The residue was dissolved in
dichloromethane (500 mL) and then chloroiodomethane (72.8 mL, 1 mol) was added
and the solution was stirred for 6h at room temperature. The solution was
to concentrated under reduced pressure and the residue was shaken with hexane
/ ethyl
acetate ( 1:1 v/v, 400 mL). The yellow crystalline solid was filtered off and
the filtrate
was washed with aqueous solution of sodium thiosulfate (0.1 M) and then
filtered
through anhydrous sodium sulfate and evaporated to dryness. The residue was
column chromatographed (silica gel, 1-2% MeOH in CHZCIz), to give 20.8 g ofN-
BOC-L-isoleucine chloromethyl ester.
b) N-BOC-L-isoleucine iodomethyl ester.
To a solution ofN-BOC-L-isoleucine chloromethyl ester (19.6 g, 70 mmol) in
acetonitrile (300 mL), was added sodium iodide (31.5 g, 210 mmol). The
solution
2o was stirred for 4 h at 60 °C. The resulting suspension was filtered
and the filtrate was
evaporated. The residue was dissolved in CHzCl2 and washed with aqueous sodium
thiosulfate (0.1 M}. The organic phase was dried (NaZS04) and concentrated
under
reduced pressure. The crude product was column chromatographed (silica gel, 2%
MeOH in CHZCIz), to give 22.6 g of N-BOC-L-isoleucine iodomethyl ester.
'H-NMR (CDC13): 6.04 (d, 1H), 5.82 (d, 1H), 4.97 (d, 1H), 4.25 (dd, 1H), 1.98-
1.80
(m, 1H), 1.43 (s, 9H), 1.50-1.05 (m, 2H), 0.97-0.88 (m, 6H).
Example PI-2
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49
2.2-dimethvl-3-lN-Boc-L-valvloxy~propionic acid iodomethyl ester
O
O NH
I~0~~~
OO
a) 2,2-dimethyl-3-( N-Boc-L-valyloxy)propionic acid.
N-Boc-L-valine (I0.8g, 50 mmoie), 4-dimethylaminopyridine (610 mg, 5 mmoIe)
and DCC (6.18 g, 30 mmole) were dissolved in methylene chloride (100 ml).
After
stirring for 2 hour the mixture was filtered. To the filtrate were added 2,2-
dimethyl-3-
hydroxy-propionic acid (3.54g, 30 mmoIe) and pyridine (10 m1). After 18 hr,
the
reaction mixture was filtered, and the filtrate was poured into sodium
hydrogen
carbonate aqueous solution, the organic phase was then washed with citric acid
to aqueous solution and water succesively. After evaporation the product was
isolated
with silica gel column chromatography to yield 4.4g.
b) 2,2-dimethyl-3-(N-Boc-L-valyloxy)propionic acid chloromethyl ester.
2,2-Dimethyl-3-(N-Boc-L-valyloxy)propionic acid (3.9 g, 12.3 mmole) was
~5 dissolved in dioxane (60 m1). To the solution was added tetrabutylammonium
hydroxide aqueous solution (40 %, 7.78 ml, 12 mmole). The solution was dried
in
vacuo, and it was coevaporated with toluene for several times. The residue was
dissolved in methylene chloride and then chloroiodomethane (18.9 ml, 260
mmole)
was added to the solution. After 18 hr, the reaction solution was evaporated
and the
20 product was isolated with silica gel column chromatography to yield 3.7 g.
c) 2,2-dimethyl-3-(N-Boc-L-valyloxy)propionic acid iodomethyl ester.
2,2-Dimethyl-3-( N-Boc-L-valyloxy)propionic acid chloromethyl ester ( 3.6 g,
10
mmole ) was dissolved in acetonitrile (50 ml). Sodium iodide (2.1 g, 14 mmole)
was
25 added to the solution. After reaction at 70° C for 2 hr, the
reaction mixture was
filtered and the residue was dissolved in methylene chloride (20 ml) and
refiltered.
The solution was dried and gave 4.348 of the titled product..
'H-NMR (CDCl3): 5.92 (dd, 2H) 5.10 (d, 1H) 4.24 (m, 1H) 4.15 (dd, 2H) 2.01
30 (m, 1 H) 1.44 (s, 9H) 1.25 (d, 6H) 0.91 (m, 6H )
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WO 99/51613 PCT/SE99/00528
Example P-I-3
3,3- bis fN-CBz-L-valyloxvmeth~)-propionic acid iodometh 1 ester
O
N O ~ I
O
I~o
O O
o ~
0
0
a) 3,3-Bis (N-CBz-L-valyloxymethyl)-propionic acid chloromethyl ester.
3,3- bis (N-CBz-L-valyloxymethyl)-propionic acid (3 g, 5 mmole) prepared
according to PCT/SE99/00194) was dissolved in dioxane (20 ml). To the solution
was added tetrabutylammonium hydroxide aqueous solution (40 %, 3.11 ml, 4.8
to mmole). The solution was dried in vacuo, and it was coevaporated with
toluene
several times. The residue was dissolved in methylene chloride (15 ml) and
then
chloroiodomethane (7.3 ml, 100 mmole) was added to the solution. The reaction
solution was refluxed for 18 hr and then evaporated and the product was
isolated
with silica gel column chromatography. 900 mg.
b) 3,3-bis-(N-CBz-L-valyloxymethyl)propionic acid iodomethyl ester .
3,3-Bis (N-CBz-L-valyloxymethyl)-propionic acid chloromethyl ester (900 mg,
1.38
mmole) was dissolved in acetonitrile (S ml). Sodium iodide (289 mg, 1.93
mmole)
was added to the solution. After reaction at 70° C for 3 hr, the
reaction mixture was
2o filtered and the residue was dissolved in methylene chloride (5 ml) and
refiltered.
The solution was dried and gave the titled product. 800 mg.
'H-NMR (CDC13): 7.35 (m, 10 H) 5.88 (dd, 2H) 5.25 (d, 2H) 4.29 (m, 2H) 4.18
(m,
4H) 2.56 (m, 1H) 2.42 (d, 2H) 2.16 (m, 2H) 0.93 (m 12 H)
Example P-I-4
2-lN-CBz-L-valyloxylethoxycarbonyloxymethyl iodide
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S1
O
O
p N"O w
O~ ~O
O
2-(N-CBz-L-valyloxy)ethoxycarbonyloxymethyl chloride (1.16 g, 3 mmole) was
dissolved in acetonitrile (10 ml). Sodium iodide (630 g, 4.2 mmole) was added
to the
solution. After reaction at 65° C for 2.5 hr, the reaction mixtwe was
cooled down to
room temperature and filtered and the residue was dissolved in methylene
chloride (5
ml) and refiltered. The solution was dried and gave the titled product. 1.2 g.
'H-NMR (CDCl3): 7.35 (m, SH) 5.93 (dd, 2H) 5.26 (d, 1H) 5.11 (s, 2H) 4.39 (m,
SH)
1 o 2.18 (m, 1 H) 0.94 (m, 6 H).
Example P-I-S
1,3-bisfN-tert-butoxvcarbonyl-L-valyloxy)-2-propyl iodomethYl carbonate
O
N-Boc-Valyl -O
~--O O~I
N-Boc-Valyl -O
15 a) 1-O-(N tent-butoxycarbonyl-L-valyl)glycerol.
N tert-Butoxycarbonyl-L-valine (32.53 g, 0.150 mol), N,N'-dicyclohexyl-
carbodiimide (37.85 g, 0.183 mol, and 4-dimethylaminopyridine (1.83 g, 0.015
mol)
were added to glycerol (138.12 g., 1.5 mol) in 500 mL dry DMF and the mixture
was
stirred at rt under NZ for 3 days. The reaction mixture was filtered,
concentrated under
2o vacuum, and then partitioned between 300 mL EtOAc and 150 mL HZO. The
aqueous phase was reextracted with 150 mL EtOAc. The organic phases were
combined and washed successively with 100 mL each of saturated aqueous
NaHC03, saturated NH4Cl, and brine. Drying over anhydrous NazS04, and
concentration under vacuum gave a viscous light yellow oil as crude product.
Flash
25 column chromatography on silica gel with 4/1 EtOAc - petroleum ether (BP 40-
60
°C) gave 18.27 g (42%) of product (alternative nomenclature: 3-(N-tert-
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butoxycarbonyl-L-valyloxy)-1,2-propanediol). Reactions done overnight gave
similar
yields.
b) 1,3-di-O-(N tert-butoxycarbonyl-L-valyl)glycerol.
1-O-(N tert-butoxycarbonyl-L-valyl)glycerol (17.95 g. 61.6 mmol), Boc-L-valine
(6.69 g, 30.8 mmol), DMAP (0.38 g, 3.1 mmol}, and DCC (7.10 g, 34.4 mmol) in
240 mL CHZCIZ and 60 mL DMF were stirred at rt under Nz for 18 h. The reaction
mixture was filtered, concentrated under vacuum, and redissolved in 200 mL
EtOAc.
The organic solution was washed with 50 mL saturated NH4C1. The aqueous phase
was reextracted with SO mL EtOAc. The organic phases were combined, washed
successively with 50 mL saturated NaHC03 and 50 mL brine, dried over NazS04,
and
concentrated under vacuum. Flash column chromatography of the crude material
on
silica gel (eluent 2/ 1 petroleum ether - EtOAc, and then EtOAc) gave 7.41 g
(49%)
of the title compound (alternative nomenclature: 1,3-bis(N-tert-butoxycarbonyl-
L-
valyloxy)-2-propanol).
c) 2-D-chloromethoxycarbonyl-1,3-di-O-(N tert-butoxycarbonyl-L-valyl)glycerol
Chloromethyl chloroformate (2.70 mL, 30 mmol) was added to a solution of 1,3-
di-
O-(N tert-butoxycarbonyl-L-valyl)glycerol (7.27 g, 14.8 mmol) and pyridine
(7.2
mL, 89 mmol) in 60 mL dry CHZC12, in an ice bath, under N2. After stirring for
1 h
45 min, the reaction mixture was diluted with 100 mL CHZCIz and washed with 40
mL water. The aqueous phase was reextracted with 20 mL H20. The organic phases
were combined, washed with 40 mL saturated NaHC03, followed by 2 x 50 mL
brine, dried over NazS04, and concentrated under vacuum. Flash column
chromatography on silica gel with 2/1 hexane- EtOAc gave 8.03 g (93%) of the
title
compound (alternative nomenclature: 1,3-bis(N-tert-butoxycarbonyl-L-valyloxy)-
2-
propyl chloromethyl carbonate).
d) 2-O-iodomethoxycarbonyl-1,3-di-O-(N tert-butoxycarbonyl-L-
valyl)glycerol.
3o A solution of 2-O-chloromethoxycarbonyl-1,3-di-O-(N tert-butoxycarbonyl-L-
valyl)propane-1,2,3-triol (7.86 g, 13.5 mmol} and NaI (8.09 g, 54.0 mmol) in
135 mL
dry acetonitirile was refluxed at 80 °C for 4 h under N,. The reaction
mixture was
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concentrated under vacuum, and then partitioned between 150 mL diethyl ether
and
SO mL HZO. The aqueous layer was reextracted with 2 x 25 mL ether. The
combined
organic phases were washed successively with 25 mL aqueous NazS2O3 and 50 mL
brine, dried over NazS04, and concentrated. Flash column chromatography
(silica
gel, 2/1 hexane-ethyl acetate gave 8.38 g (92%) title product (alternative
name: 2-
iodomethoxycarbonyloxy-1,3-bis-(N-tert-butoxycarbonyl-L-valyloxy)propane or
1,3-
bis(N-tert-butoxycarbonyl-L-valyloxy)-2-propyl iodomethyl carbonate) as a
white
solid.
'H NMR (250 MHz, CDCl3) S 0.81 (d, 6H), 0.88 (m, 6H), 1.36 (s, 18H), 2.06 (m,
2H), 4.11-4.46 (m, 6H), 5.0 (br d, 2H), 5.12 (m, 1 H), 5.88 (s, 2H).
Example P-1-6
3-benzvloxvcarbon~propionic acid iodomethyl ester
a) Succinic acid monobenzyl ester .
Succinic anhydride (30 g, 300 mmole) was dissolved in methylene chloride (300
ml).
To the solution were added benzyl alcohol (10.2 ml, 100 mmole), 4-
dimethylaminopyridine (1.22 g, 10 mmole) and pyridine (48 ml). After 3 hours
the
reaction mixture was poured in to citric acid aqueous solution. The organic
phase
was concentrated to small volume and sodium hydrogen carbonate and water were
added. Then mixture was stirred for 30 min. The aqueous phase was collected,
and
to it was added citric acid aqueous solution. The product precipitated out,
was
collected and dried. 15.3 g.
b) 3-benzyloxycarbonylpropionic acid chloromethyl ester.
Succinic acid monobenzyl ester (4.16 g, 20 mmole) was dissolved in dioxane (20
ml). To the solution was added tetrabutylammonium hydroxide aqueous solution
(40
%, 11.6 ml, 18 mmole). The solution was dried in vacuo and coevaporated with
toluene several times. The residue was dissolved in methylene chloride (60 ml)
and
then chloroiodomethane (14.5 ml, 200 mmole) was added to the solution. The
reaction solution was stirred for 18 hr and then evaporated and the product
was
isolated with silica gel column chromatography. 3.64 g
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'H-NMR (CDC13): 7.35 (m, SH), 5.67 (s, 2H), 5.13 (s, 2H), 2.72 (s, 4H).
c) 3-Benzyloxycarbonylpropionic acid iodomethyl ester.
3-Benzyloxycarbonylpropionic acid chloromethyl ester (2 g, 1.38 mmole) was
dissolved in acetonitrile (30 ml). Sodium iodide (1.6 g, 10.9 mmole) was added
to
the solution. After reaction at 70° C for 3 hr, the reaction mixture
was filtered and the
residue was dissolved in methylene chloride (20 ml) and refiltered. The
solution was
dried and gave intermediate 3-benzyloxycarbonylpropionic acid iodomethyl ester
in
to quantitative yield. This intermediate is bonded to an accessible function
of a drug,
such as a ring hydroxy or carboxy function using conventional
alkylation/acylation
conditions as described generally herein. Following deprotection of the
terminal
carboxy, a di/trifunctional linker bearing R2, such as 1,3-bis- O-(L-
valyl)glycerol or
iodomethyloxy-L-valyl is acylated/alkylated thereon or RZ amide bonded thereon
by
15 conventional techniques as described herein, such as with DCC coupling
agent.
A-I-1
Iodomethvl 2-methyl-2-(N-benzvloxycarbonyl-L-va~loxvmeth~l propionate
2o a) 4-Methoxybenzyl 2-(hydroxymethyl)-2-methyl propionate.
2-(Hydroxymethyl)-2-methyl propionic acid was esterified by alkylation with 4-
methoxybenzyl chloride by conventional means, namely treatment with aqueous
NaOH, followed by evaporation and dissolution in an organic solvent such as
DMF
to which the 4-methoxybenzyl chloride is added and the reaction warmed and
25 agitated, such as stirring at 60 C for one hour. The reaction mixture is
cooled,
concentrated by rotavapor and the resulting concentrated suspension
partitioned
between water and dichloromethane. The organic phase is evaporated and the
residue
subjected to silica gel column chromatography, for example with 0, 2, 4% EtOH
in
dichloromethane to yield the title compound (7.10 g). Rf (2%MeOH/CHCl3) 0.40.
b) 4-Methoxybenzyl 2-(N- benzyloxycarbonyl-L-valyloxymethyl)-2-
methyl propionate.
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4-Methoxybenzyl 2-(hydroxymethyl)-2-methyl propionate (2.50 g, 10.5 mmol), N-
benzyloxy carbonyl-L-valine (2.SI g, 10 mmole), 4-dimethylaminopyridine (183
mg)
and I-hydroxybenzotriazole (l.3Sg, 10 mmole) were mixed and dissolved in N,N-
dimethylformamide (90 ml). Then dicyclohexyl-carbodiimide (2.47 g 12 mmol) was
S added. After stirring for 3 days at room temperature the suspension was
filtered and
the filtrate evaporated in vacuo. The residue was partitioned between O.1M
citric acid
and dichloromethane. The organic phase was then extracted with aqueous
saturated
NaHC03 and evaporated in vacuo. The residue was silica gel column
chromatographed (0, l, 2, 3% ethanol in dichloromethane). The appropriate
fractions
to were pooled and evaporated in vacuo to give the title compound (2.72 g).
R~. (2%MeOH/CHC13) 0.75.
d) 2-(N- benzyloxycarbonyl-L-valyloxymethyl)-2-methyl propionic acid.
To a solution of 4-methoxybenzyl 2-(N- benzyloxycarbonyl -L-valyloxymethyl)-2-
1S methyl propionate (2.72 g, 5.76 mmole), was added trifluoroacetic acid
(11.5 ml) and
the emerging dark red solution was stirred for 30 min at room temperature. The
solution was evaporated to dryness with dioxane and toluene. The residue was
silica
gel column chromatographed (2, 3, 4% ethanol in dichloromethane). The
appropriate
fractions were pooled and evaporated in vacuo to give the title compound (1.86
g). Rf
20 (2%MeOH/CHCl3) 0.30.
'H-NMR (CDCI3): 7.32 (s, SH), 5.32 (d, 1H), 5.10 (s, 2H), 4.32 (d,d, IH), 4.21
(d,d,
2H), 2.13 (m, 1H), 1.26 (s, 3H), 1.25 (s, 3H), 0.95 (d, 3H), 0.86 (d, 3H).
25 c) Chloromethyl2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-methyl
propionate.
2-(N- benzyloxycarbonyl -L-valyloxymethyl)-2-methyl propionic acid was
esterified
by conventional techniques, namely dissolution in an organic solvent such as
dioxane
and dropwise addition of aqueous tetrabutylammonium hydroxide, followed by
3o evaporation. The residue is dissolved in dichloromethane and then
chloroiodomethane and the mixture stirrred for 6 hours at room temperature,
followed by partition, shaking the filtrate with aqueous sodium thiosulphate.
O.1M,
filtration and evaporation. The title compound (1.40 g) was obtained after
silica gel
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column chromatography (0, 1, 2, 3% ethanol in dichloromethane). Rr
(2%MeOH/CHC13) 0.80.
c) Iodomethyl 2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-methyl
propionate.
Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-methyl propionate was
converted to iodide by conventional techniques, namely addition to NaI in
acetonitrile, stirring and heating, for instance to 75 C for four hours. The
resulting
suspension is filtered and the filtrate evaporated, dissolved in organic
solvent such as
to toluene and shaken with aqueous sodium thiosulphate (O.1M) and evaporation
to give
the title compound (1.25 g) practically pure. Rf (2%MeOH/CHC13) 0.80.
'H-NMR (CDCl3): 7.35 (s, SH); 5.90 (d,d, 2H), 5.24 (d, 1H), 5.10 (s, 2H), 4.31
(d,d,
1H), 4.14 (d,d, 2H), 2.16 (m, 1H), 1.22 (s, 6H), 0.96 (d, 3H), 0.87 (d, 3H).
Example A-I-2
Iodomethvl 2-(N-benzvloxycarbonyl-L-valyloxyl-DL propionate
O CH3
O O~I
CBzNH
a) Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxy)-DL-propionate.
2-(N-benzyloxycarbonyl-L-valyloxy) propionic acid (1 g) was esterified by the
method described in Example A-I-I, step d. The title compound (0.76 g) was
obtained after silica gel column chromatography (0, 1 % ethanol in
dichloromethane).
Rf (2%MeOH/CHC13) 0.75.
b) Iodomethyl2-(N-benzyloxycarbonyl-L-valyloxy)-DL-propionate.
Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-methyl propionate was
converted to iodide by the method described in Example A-I-1, step a to give
the title
compound (0.95 g) practically pure. Rf (2%MeOH/CHC13) 0.75.
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'H-NMR (CDC1,): 7.33 (s, SH), 5.98 (d, 1H), 5.86 (d, 1H), 5.26 (d, 1H), 5.10
(s, 2H),
5.07 (q, 1H), 4.38 (d,d, 1H), 2.30 (m, 1H), 1.49 (d, 3H), 1.03 (d, 3H), 0.95
(d, 3H).
Example A-I-3
Iodomethvl (1,3-di-(N-benzvloxvcarbonyl~L-valyloxy,Lpr~vl carbonate
O
N-CBz-Valyl -O ~
O- _O~I
N-
CBz-Vatyl-O
a) Chloromethyl (1,3-di-(N-benzyloxycarbonyl)-L-valyloxy)-2-propyl
carbonate.
1o To a solution of 1,3-di-((N-benzyloxycarbonyl)-L-valyloxy)propan-2-of (1.34
g, 2.4
mmole) in dichloromethane (10 ml) was added dry pyridine (1.15 ml, 14.4 mmol)
and chloromethyl chloroformate (0.43 ml, 4.8 mmole) at 0°C. The
reaction was then
stirred for 30 min and then poured into aqueous 50% saturated sodium chloride
/
O.1M citric acid solution and extracted with dichloromethane. The organic
phase was
evaporated and the residue silica gel column chromatographed (0, 1, 1.5%
ethanol in
dichloromethane). The appropriate fractions were pooled and evaporated in
vacuo to
give the title compound (1.26g). Rf (2%MeOH/ CHC13) 0.85.
2o b) Preparation of iodomethyl (1,3-di-(N-benzyloxycarbonyl)-L-valyloxy)-
2-propyl carbonate.
Chloromethyl (1,3-di-(N-benzyloxycarbonyl)valyloxy)-2-propyl carbonate was
converted to iodide by the method described in Example A-I-l, step e) to give
the
title compound (1.37 g) practically pure. Rr (2%MeOH/CHC13) 0.85.
'H-NMR (CDC13): 7.34 (s, lOH), 5.93 (d, 1H), 5.89 (d, 1H), 5.21 (m, 3H), 5.11
(s,
4H), 4.50-4.17 (m, 6H), 2.12 (m, 2H), 0.97 (d, 6H), 0.88 (d, 6H).
Example A-I-4
3o Iodomethvl2-lN-benzyloxycarbonyl-L-valyloxy~isobutyrate
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O
CBzNH O O~I
O
a) 4-Methoxybenzyl 2-hydroxyisobutyrate.
2-hydroxy isobutyric acid (1.56 g) was esterified by alkylation with 4-
methoxybenzyl chloride by the method described in Example A-I-1, step a). The
title
s compound (2.65 g) was obtained after silica gel column chromatography (0, 1,
2%
ethanol in dichloromethane). Rf (2%MeOH/CHCI3) 0.45.
b) 4-Methoxybenzyl 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyrate.
4-methoxybenzyl 2-hydroxyisobutyrate was acylated with N-benzyloxycarbonyl-L-
to valine by the method described in Example A-I-1, step b). The title
compound (3.21
g) was obtained after silica gel column chromatography (0; 1, 1.5% ethanol in
dichloromethane). R~ (2%MeOH/CHCl3) 0.70.
c) 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyric acid.
1 s 4-methoxybenzyl 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyrate was de-
esterified
by the method described in Example A-I-1 step c. The title compound (2.01 g)
was
obtained after silica gel column chromatography (2, 10, 20% ethanol in
dichloromethane). Rf (2%MeOH/CHCI,) 0.30.
20 'H-NMR (CDCI3): 7.32 (s, SH), 5.33 (d, 1H), 5.10 (s, 2H), 4.31 (d,d, 1H),
2.22
(m, 1 H), 1.57 (s, 6H), 0.98 (d, 3H), 0.89 (d, 3H).
d) Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyrate.
2-(N-benzyloxycarbonyl-L-valyloxy) isobutyric acid was esterified by the
method
2s described in Example A-I-1, step d. The title compound (1.90 g) was
obtained after
silica gel column chromatography (0, 1, 1.5% ethanol in dichloromethane). Rf
(2%MeOH/CHCI3) 0.80.
e) Iodomethyl 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyrate.
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Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxy) isobutyrate was converted to
iodide by the method described in Example A-I-l, step a to give the title
compound
(2.32 g) practically pure. Rf (2%MeOH/CHC13) 0.80.
'H-NMR (CDC13): 7.33 (s, SH), 5.89 (s, 2H), 5.22 (d, 1H), 5.11 (s, 2H), 4.29
(d,d,
1H), 2.21 (m, 1H), 1.55 (s; 3H), 1.53 (s, 3H), 1.00 (d, 3H), 0.93 (d, 3H).
EXAMPLE A-I-5
Iodomethvl 2-lN-benzvloxvcarbonyl-L-valvloxv)-3-methyl-(S)l+ -butyrate
O
CBzNH O O~I
O
io
a) 4-Methoxybenzyl 2-hydroxy-3-methyl-(S)-(+)-butyrate.
2-hydroxy-3-methyl-(S)-(+)-butyric acid (1.77 g) was esterified by alkylation
with 4-
methoxybenzyl chloride by the method described in Example A-I-1, step a. The
title
compound (3.10 g) was obtained after silica gel column chromatography (0, 1,
2%
~5 ethanol in dichloromethane). Rf (2%MeOH/CHC13) 0.50.
b) 4-Methoxybenzyl2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-
(+)-butyrate.
20 4-Methoxybenzyl 2-hydroxy-3-methyl-(S)-(+)-butyrate was acylated with N-
benzyloxycarbonyl-L-valine by the method described in Example A-I-1, step b.
The
title compound (5.74 g) was obtained after silica gel column chromatography
(0, 1,
1.5% ethanol in dichloromethane). Rf (2%MeOH/CHCI,) 0.70.
25 c) 2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-butyric acid.
4-methoxybenzyl 2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-butyrate
was de-esterified by the method described in Example A-I-1, step c. The title
compound (3.41 g) was obtained after silica gel column chromatography (2, 10,
20%
ethanol in dichloromethane). Rr (2%MeOH/CHC13) 0.45.
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'H-NMR (CDC13): 7.36 (s, SH), 5.38 (d, 1H), S.I 1 (s, 4H), 4.90 (d, 1H), 4.4I
(d,d,
1H), 2.28 (m, 2H), 1.04-0.89 (m, 12H).
d) Chloromethyl2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-
butyrate.
2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-butyric acid was
esterified by
the method described in Example A-I-1, step d. The title compound (2.96 g) was
obtained after silica gel column chromatography (0, 1, 2% ethanol in
dichloromethane). Rf (2%MeOH/CHC13) 0.85.
e) Iodomethyl 2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-
butyrate.
Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-butyrate was
converted to iodide by the method described in Example A-I-1, step a to give
the title
compound (3.64 g) practically pure. Rf (2%MeOH/CHC13) 0.85.
'H-NMR (CDC13): 7.36 (s, SH), 6.00 (d, 1H), 5.83 (d, 1H), 5.28 (d, 1H), 5.11
(s, 4H),
4.83 (d, 1H), 4.41 (d,d, 1H), 2.29 (m, 2H), 1.05-0.90 (m, 12H).
2o EXAMPLE A-I-6
Iodomethvl 2-(N-benzyloxycarbonvl-L-va~Ioxy)-2=phenyl-DL-acetate
O
cBZNH ° o~~
0
I
a) 4-Methoxybenzyl 2-hydroxy-2-phenyl-DL-acetate.
DL-mandelic acid (2.28 g) was esterified by alkylation with 4-methoxybenzyl
chloride by the method described in Example A-I-1, step a. The title compound
(3.69
g) was obtained after silica gel column chromatography (0, 1, 1.5% ethanol in
dichloromethane). Rf (2%MeOH/CHCl3) 0.55.
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b) 4-Methoxybenzyl2-(N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-
acetate.
4-Methoxybenzyl 2-hydroxy-2-phenyl-DL-acetate was acylated with N-
benzyloxycarbonyl-L-valine by the method described in Example A-I-l, step b.
The
title compound (6.50 g) was obtained after silica gel column chromatography
(0, 1,
1.5% ethanol in dichloromethane). Rf (2%MeOH/CHCI,) 0.75.
c) 2-{N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-acetic acid.
4-Methoxybenzyl 2-(N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-acetate was
to de-esterified by the method described in Example A-I-1, step c. The title
compound
{4.75 g) was obtained after silica gel column chromatography (2, 10, 20%
ethanol in
dichloromethane). Rf (2%MeOH/CHCl3) 0.40.
'H-NMR (CDC13): 7.36 (m, lOH), 5.91 (d, 1H), 5.27 (m, 1H), 5.04 (s, 2H), 4.57-
4.40
(2xd,d, 1H), 2.30 (m, 1H), 1.01-0.82 (m, 6H).
d) Chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-
acetate.
2-{N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-acetic acid was esterified by
the
method described in Example A-I-1, step d. The title compound (1.69 g) was
obtained after silica gel column chromatography (0, 1, 2% ethanol in
dichloromethane). R,. (2%MeOH/CHCl3) 0.80.
e) Iodomethyl2-(N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-acetate.
Chloromethyl 2-{N-benzyloxycarbonyl-L-valyloxy)-2-phenyl-DL-acetate was
converted to iodide by the method described in Example A-I-1, step a to give
the title
compound (1.89 g) practically pure. R~ (2%MeOH/CHC13) 0.80.
'H-NMR {CDCl3): 7.36 (m, lOH), 5.94-5.82 (m, 3H), 5.28 (m, 1H), 5.10 (s, 2H),
4.46 (m, 1 H), 2.21 (m, 1 H), 1.08-0.8 S (m, 6H).
Example A-I-7
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Iodomethvl 4-fN-benz r~loxycarbonyl-L-valvloxy) benzoate
O
CBzNH ~
O ~. I O.~ I
i
O
a) 4-Methoxybenzyl 4-hydroxybenzoate.
4-Hydroxybenzoic acid (1.38 g) was esterified by alkylation with 4-
methoxybenzyl
chloride by the method described in Example A-I-1, step a. The title compound
(2.06
g) was obtained after silica gel column chromatography (0, 1, 2, 3% ethanol in
dichloromethane). Rf (2%MeOH/CHC13) 0.40.
b) 4-Methoxybenzyl 4-(N-benzyloxycarbonyl-L-valyloxy) benzoate.
4-Methoxybenzyl 4-hydroxybenzoate was acylated with N-benzyloxycarbonyl-L-
valine by the method described in Example A-I-1, step b. The title compound
(2.71
g) was obtained after silica gel column chromatography {0, 1 % ethanol in
dichloromethane). Rf (2%MeOH/CHCl3) 0.70.
c) 4-(N-benzyloxycarbonyl-L-valyloxy) benzoic acid.
4-Methoxybenzyl 4-(N-benzyloxycarbonyl-L-valyloxy) benzoate was de-esterified
by the method described in Example A-I-1, step c. The title compound (1.86 g)
was
obtained after silica gel column chromatography (2, 10, 20% ethanol in
dichioromethane). Rf (2%MeOH/CHCl3) 0.20.
'H-NMR (CDC13): 8.15 (d, 2H), 7.34 (m, SH), 7.22 {d, 2H), 5.38 (d, 1H), 5.17
(s,
2H), 4.58 (d,d, 1H), 2.34 (m, 1H), 1.12 (s, 3H), 0.96 (d, 3H).
d) Chloromethyl 4-(N-benzyloxycarbonyl-L-valyloxy) benzoate.
4-(N-benzyloxycarbonyl-L-valyloxy) benzoic acid was esterified by the method
described in Example A-I-l, step d. The title compound (0.95 g) was obtained
after
silica gel column chromatography (0, 1 % ethanol in dichloromethane). Rf
(2%MeOH/CHC13) 0.80. -
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e) Iodomethyl 4-(N-benzyloxycarbonyl-L-valyloxy) benzoate.
Chloromethyl 4-(N-benzyloxycarbonyl-L-valyloxy) benzoate was converted to
iodide by the method described in Example A-I-1, step a to give the title
compound
(1.16 g) practically pure. R,. (2%MeOH/CHC13) 0.80.
'H-NMR (CDC13): 8.11 (d, 2H), 7.35 (m, SH), 7.21 (d, 2H), 6.15 (s, 2H), 5.32
(d,
1H), 5.14 (s, 2H), 4.55 (d,d, 1H), 2.34 (m, 1H), 1.10 (s, 3H), 1.03 (d, 3H).
Example A-1-8
to Iodomethyl5-(N-CBz-L-valvloxy~-2,2-dimethylvalerate
O
I ~ O O. Cbz-Val
a) 4-Methoxybenzyl 2,2-dimethyl-4-pentenoate
To a solution of 2,2-dimethyl-4-pentenoic acid (11.5 g, 90 mmol) in DMF (250
mL)
at room temperature, was added potassium tent-butoxide ( 11.1 g, 99 mmol). The
15 reaction mixture was stirred at 60 °C for lh. 4-Methoxybenzyl
chloride (16.9 g, 108
mmol) was added and the reaction mixture was stirred at 60 °C for 4h.
The DMF was
evaporated under vacuum, the residue was dissolved in ether (S00 mL) and
washed
with water (3 x 200 mL). The organic phase was dried with NazS04 and
evaporated
to give 21.4 g of 4-methoxybenzyl 2,2-dimethyl-4-pentenoate.
b) 4-Methoxybenzyl5-hydroxy-2,2-dimethylvalerate.
A mixture of 4-methoxybenzyl 2,2-dimethyl-4-pentenoate (9.50 g, 38 mmol) and 9-
BBN (115 mL, 57 mmol, 0.5 M in THF) was stirred at 60 °C for 60 min,
whereupon
the reaction mixture was cooled to -5 °C. Hz0 (35 mL) was added, the
reaction
mixture was stirred for 5 min at -S °C, an aqueous solution of NaOH (35
mL, 3M)
was added and the reaction mixture was stirred for a further 10 min at -5
°C. An
aqueous solution of H202 (35 mL, 30%) was added dropwise and the temperature
of
the reaction mixture was allowed to assume room temperature, whereupon the
reaction mixture was stirred for 30 min at room temperature. After
evaporation, water
(200 mL) was added and the resulting mixture was extracted with CHzCl2 (5 x
200
mL). The combined organic layers were dried (NaZS04) and concentrated under
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reduced pressure. The crude product was column chromatographed (silica gel,
1-->8% MeOH in CHzCIZ), to give 6.32 g of 4-methoxybenzyl 5-hydroxy-2,2-
dimethylvalerate.
c) 4-Methoxybenzyl5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate.
To a mixture of DCC (9.41 g, 46 mmol), DMAP (0.586 g, 4.8 mmol) and N-CBz-L-
valine (12.1 g, 48 mmol) in CHZCIz (200 mL) at 0 °C, was added dropwise
a solution
of 4-methoxybenzyl 5-hydroxy-2,2-dimethyl-valerate (6.40 g, 24 mmol) in CHZC12
(50 mL). After 1 h at 0 °C, the temperature of the reaction mixture was
allowed to
1o assume room temperature and then the mixture was stirred for Sh at room
temperature. The mixture was filtered through a glass filter and the solvent
was
removed under reduced pressure. The crude product was column chromatographed
(silica gel, 1-~4% MeOH in CH~CIZ), to give 8.61 g 4-methoxybenzyl 5-(N-CBz-L-
valyloxy)-2,2-dimethylvalerate.
d) 5-(N-CBz-L-valyloxy)-2,2-dimethylvaleric acid.
To a solution of 4-methoxybenzyl 5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate
(8.24
g, 16.5 mmol) in CHZCIz (100 mL) at room temperature, was added
trifluoroacetic
acid (5 mL). After lh at room temperature, the reaction mixture was
concentrated
2o under reduced pressure. The crude product was column chromatographed
(silica gel,
3~5% MeOH in CHZC12), to give 6.00 g of 5-(N-CBz-L-valyloxy)-2,2-
dimethylvaleric acid.
'H-NMR (CDC13): 10.94 (br s, 1H), 7.35 (s, SH), 5.45 (d, 1H), 5.11 (s, 2H),
4.30
(dd, 1 H), 4.21-4.00 (m, 2H), 2.28-2.07 (m, 1 H), 1.68-1.51 (m, 4H), 1.21 (s,
6H), 0.97
(d, 3H), 0.89 (d, 3H).
e) Chloromethyl 5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate.
To a solution of 5-(N-CBz-L-valyloxy)-2,2-dimethylvaleric acid (5.88 g, 15.5
mmol)
3o in dioxane (100 mL), was added dropwise a 40% aqueous solution of
tetrabutylammonium hydroxide (10.1 g). After stirnng for S min, the solution
was
evaporated to dryness through co-evaporation with dioxane and toluene. The
residue
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was dissolved in dichloromethane (100 mL) and then chloroiodomethane (11.3 mL,
155 mmol) was added and the solution was stirred for 6h at room temperature.
The
solution was concentrated under reduced pressure and the residue was shaken
with
hexane / ethyl acetate (1:1 v/v, 200 mL). The yellow crystalline solid was
filtered off
5 and the filtrate was washed with aqueous solution of sodium thiosulfate (0.1
M) and
the filtered through anhydrous sodium sulfate and evaporated to dryness. The
residue
was column chromatographed (silica gel, 1-4% MeOH in CHZCIz), to give 3.95 g
of
chloromethyl 5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate.
l0 f) Iodomethyl5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate.
To a solution of chloromethyl 5-(N-CBz-L-valyloxy)-2,2-dimethylvalerate (3.85
g, 9
mmol) in acetonitrile (SO mL), was added sodium iodide (5.40 g, 36 mmol). The
solution was stirred for 4 h at 60 °C. The resulting suspension was
filtered and the
filtrate was evaporated. The residue was dissolved in CHZCIz and washed with
15 aqueous sodium thiosulfate (0.1 M). The organic phase was dried (Na2S04)
and
concentrated under reduced pressure. The crude product was column
chromatographed (silica gel, 1% MeOH in CHZC12), to give 4.26 g of iodomethyl
5-
(N-CBz-L-valyloxy)-2,2-dimethylvalerate
20 'H-NMR (CDC13): 7.34 (s, SH), 5.90 (s, 2H), 5.32 (d, 1H), 5.10 (s, 2H),
4.29 (dd,
1H), 4.18-4.02 (m, 2H), 2.26-2.08 (m, 1H), 1.65-1.50 (m, 4H), 1.17 (s, 6H),
0.97 (d,
3H), 0.89 (d, 3H).
Example A-1-9
25 2-(N-CBz-L-valyloxY)-ethyl iodomethyl carbonate
O
I~O~O~O ~N-Cbz)-Val
a) 2-(N-CBz-L-valyloxy)-ethanol
To a mixture of DCC (11.4 g, 55 mmol), DMAP (0.611 g, 5 mmol) and
ethyleneglycol (55.8 mL, 1 mol) in CH2Cl2 (300 mL) at 0 °C, was added
dropwise a
3o solution ofN-CBz-L-valine (12.6 g, 50 mmol) in CHZC12 (100 mL). After lh at
0 °C,
the temperature of the reaction mixture was allowed to assume room temperature
and
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then the mixture was stirred for Sh at room temperature. The mixture was
filtered
through a glass filter and the solvent was removed under reduced pressure. The
crude
product was column chromatographed (silica gel, 5-X10% MeOH in CHZC12), to
give
12.0 g 2-(N-CBz-L-valyloxy)-ethanol.
b) 2-(N-CBz-L-valyloxy)-ethyl chloromethyl carbonate.
To a mixture of 2-(N-CBz-L-valyloxy)-ethanol (12.0 g, 40.6 mmol) and pyridine
(19.7 mL, 0.24 mmol) in CHzCl2 (300 mL) at 0 °C, was added dropwise
chloromethyl chloroformate (I0.5 g, 81.2 mmol). After 30 min at 0 °C,
the reaction
1 o mixture was washed with H20 (200 mL). The H20 phase was washed with CI
IZCIz
(100 mL) and the solvent of the combined organic phases was removed under
reduced pressure. The crude product was column chromatographed (silica gel,
0.5-~ 1 % MeOH in CHZCl2), to give 8.26 g 2-(N-CBz-L-valyloxy)-ethyl
chloromethyl carbonate.
1s
c) 2-(N-CBz-L-valyloxy)-ethyl iodomethyl carbonate.
To a solution of 2-(N-CBz-L-valyloxy)-ethyl chloromethyl carbonate (3.88 g, 10
mmol) in acetonitrile (50 mL), was added sodium iodide (7.50 g, 50 mmol). The
solution was stirred for 4 h at 60 °C. The resulting suspension was
filtered and the
20 filtrate was evaporated. The residue was dissolved in CHZCIz and washed
with
aqueous sodium thiosulfate (0.1 M). The organic phase was dried (Na2S04) and
concentrated under reduced pressure, to give 4.51 g 2-(N-CBz-L-valyloxy)-ethyl
iodomethyl carbonate.
25 'H-NMR (CDCI,): 7.34 (s, SH), 5.93 (s, 2H), 5.26 (d, 1H), 5.11 (s, 2H),
4.48-4.26
(m, SH) 2.28-2.10 (m, 1H), 0.97 (d, 3H), 0.90 (d, 3H).
Example A-I-10
2,2-dimethyl-3-f N-CBz-D-vary)-propionic acid iodomethyl ester
0
HN- _O
O
O
I~O
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a) 2,2-dimethyl-3-(N-CBz-D-valyloxy)-propionic acid.
To a solution of 2,2-dimethyl propionic acid 4-methoxybenzyl ester (4.7 g, 20
mmole) and N-CBz-D-valine (5.5 g, 22 mmole) in dichloromethane (100 ml) were
added 4-dimethyaminopyridine (305 mg, 2.5 mmole) and DCC (5.15 g, 25 mmole).
After 18 hr, the solution was washed successively with sodium bicarbonate
aqueous
solution, citric acid solution and water. The organic phase was dried and the
residue
was dissolved in dichloromethane (100 ml). To the solution was added
trifluoroacetic
acid ( 10 ml). After 3 hr, it was evaporated and the product was isolated with
silica
to gel column chromatography. 4.5 g.
'H-NMR (CDCl3): 7.36 (m, 5 H) 5.11 (s, 2H) 4.30 (m, 1H) 4.18 (dd, 2H) 2.17 (m,
1H), 1.23 (d, 6 H) 0.93 (m, 6H).
b) 2,2-dimethyl-3- ( N-CBz-D-Valyloxy )-propionic acid chloromethyl
ester.
(2,2-dimethyl-3-(N-CBz-D-valyloxy)-propionic acid (4.5 g, 12.8 mmole) was
dissolved in dioxane (20 ml). To the solution was added tetrabutylammonium
hydroxide aqueous solution (40 %, 8.3 ml, 12.8 mmole). The solution was dried
in
vacuo, and it was coevaporated with toluene several times. The residue was
dissolved
in methylene chloride and then chloroiodomethane (18 ml, 260 mmole) was added
to
the solution. After 18 hr, the reaction solution was evaporated and the
product was
isolated with silica gel column chromatography. 3.5 g.
c) 2,2-dimethyl-3-(N-CBz-D-valyloxy)-propionic acid iodomethyl ester.
2,2-Dimethyl-3-(N-CBz-D-valyloxy)-propionic acid chloromethyl ester (2.4 g, 6
mmole) was dissolved in acetonitrile (30 ml). Sodium iodide (1.26 g, 8.4
mmole)
was added to the solution. After reaction at 70° C for 2 hr, the
reaction mixture was
filtered and the residue was dissolved in methylene chloride (20 ml) and
refiltered.
3o The solution was dried and gave the titled product. 2.688.
'H-NMR (CDC13): 7.36 (m, 5 H) 5.90 (dd, 2H) 5.26 (d, 1H) 5.11 (s, 2H) 4.31
(m, 1 H) 4.15 (dd, 2H) 2.18 (m, 1 H) 1.22 (d, 6H) 0.92 (m, 6H).
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Example A-1-11
4-IN-CBz-L-valvlox~r) butyric acid iodomethyl ester
O
p HN- _O
I/~O O
O I
a) 4-(N-CBz-L-valyloxy) butyric acid t-butyl ester.
N-CBz-L-valine (16.25 g, 65 mmole) was dissolved in DMF (40 ml). To the
solution
was added potassium t-butoxide (7.24 g, 65 mmole). After 10 min, 4-
bromobutyric
acid t-butyl ester (12 g, 53 mmole) was added. The reaction mixture was kept
at 65°
C for 2.5 hr and then poured into sodium bicarbonate aqueous solution and
extracted
1o with dichloromethane. The organic phase was dried and the product was
isolated
with silica gel column chromatography. 20.1 g.
b) 4-(N-CBz-L-valyloxy)butyric acid chloromethyl ester.
4-( N-CBz-L-valyloxy) butyric acid t-butyl ester (20 g, 50.8 mmole) was
treated with
t s trifluoroacetic acid (30 ml) at 0° C for 3 h and then evaporated.
The residue was
coevaporated with toluene several time. The intermediate acid (2.56 g, 7.6
mmole)
was dissolved in dioxane ( 10 ml) and to the solution was added
tetrabutylammonium
hydroxide (40 %, 4.66 ml, 7.2 mmole). The solution was dried and dissolved in
dichloromethane (20 ml) and then chloroiodomethane (10 ml, 144 mmole) was
added
2o to the solution. After 18 hr, the reaction solution was evaporated and the
product was
isolated with silica gel column chromatography. Yield 2.1 g.
c) 4-(N-CBz-L-valyloxy)butyric acid iodomethyl ester.
4-(N-CBz-L-valyloxy) butyric acid chloromethyl ester (1.54 g, 4 mmole) was
2s dissolved in acetonitrile (15 ml). Sodium iodide (840 rng, 5.6 mmole) was
added to
the solution. After reaction at 55° C for 3 hr, the reaction mixture
was filtered and the
residue was dissolved in methylene chloride (20 ml) and refiltered. The
solution was
dried and gave the titled product. Yield 1.9 g.
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'H-NMR (CDC13): 7.36 (m, SH) 5.90 (dd, 2 H) 5.25 (d, 1H) 5.11 (s, 2H) 4.29
(dd, 1H
4.18 (t, 2H) 2.43 (t, 2H) 2.20 (m, 1H) 2.00 (m, 2H) 0.93 (dd, 6 H).
Example A-I-12
s Iodomethvl 3-(N-benzvloxvcarbon 1-L-valvloxy -benzoate
O
O
CBzNH ~ ~ 'OBI
O
a) 4-Methoxybenzyl 3-hydroxybenzoate.
To a solution of 3-hydroxybenzoic acid (6.9g, SO mmole) in DMF (100 ml) was
added potassium-tert.-butoxide (6.17 g, 55 mmole) and the mixture was stirred
at
1 o room temperature for one hour. 4-Methoxybenzyl chloride (9.4g, 60 mmole)
was
added and the mixture was stirred for 16 hours at 60°C. The mixture was
evaporated
under reduced pressure and ethyl acetate (250 ml) were added. The organic
phase
was washed five times with water, dried with sodium sulfate and evaporated
under
reduced pressure. The product was isolated by silica gel column chromatography
15 with toluene/acetone. Yield: 10.5g = 81%
b) 4-Methoxybenzyl 3-(N-benzyloxycarbonyl-L-valyloxy) benzoate.
To a cooled solution of 4-methoxybenzyl 3-hydroxybenzoate (7.7g, 29.8 mmole),
4-
dimethylaminopyridine (0.73g, 6 mmole) and N-benzyloxycarbonyl-L-valine (8.3g,
20 33 mmole) in 100 ml dichloromethane was added dicyclohexyl-carbodiimide
(7.22g,
35 mmole) and the mixture was stirred for 2 days at room temperature. The
mixture
was cooled and the urethane was filtered. The solution was evaporated and
ethyl
acetate (250 ml) was added. The organic phase was washed twice with 5% acetic
acid; 5% sodium hydrogencarbonate and water. The organic phase was dried with
25 sodium sulfate and evaporated under reduced pressure. The product was
isolated by
silica gel column chromatography with hexanelethyl acetate. Yield: 13.9g = 94%
c) 3-(N-benzyloxycarbonyl-L-valyloxy) benzoic acid.
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To a solution of 4-methoxybenzyl-3-(N-benzyloxycarbonyl-L-valyloxy)-benzoate
(13.7g, 27.8 mmole) in dichloromethane (150 ml) was added trifluoroacetic acid
(20
ml) and the mixture was stirred for 2 hours at room temperatwe. The solution
was
evaporated under reduced pressure and the product crystallized from
toluene.Yield:
s 10.1 g = 87%.
'H-NMR (DMSO d-6) 1.01 (m, 6H) 2.21 (m, 1H) 4.17 (d, d, 1H) 5.08 (s, 2H)
7.28-7.96 (m, lOH)
1o d) Chloromethyl3-(N-benzyloxycarbonyl-L-valyloxy)-benzoate.
To a solution of 3-(N-benzyloxycarbonyl-valyloxy)benzoic acid (7.42g, 20
mmole)
in 1,4-dioxane (100 ml) was added a 40% solution of tetrabutylammonium
hydroxide (12.97g, 20 mmole) and the mixture was stirred 2 hours at room
temperature. The mixture was evaporated under reduced pressure and co-
evaporated
15 two times with 1,4-dioxane and two times with toluene. The dried product
was
dissolved in dichloromethane (50 ml) and chloroiodomethane (35.3g, 200 mmole)
was added. The solution was stirred for two days at room temperature and
evaporated
under reduced pressure. Ethyl acetate (100 ml) was added and the organic phase
washed twice with water, dried with sodium sulfate and evaporated under
reduced
2o pressure: The product was isolated by silica gel column chromatography.
Yield: 3.8g
= 45%.
e) Iodomethyl 3-(N-benzyloxycarbonyl-L-valyloxy)-benzoate.
To a solution of chloromethyl 3-(N-benzyloxycarbonyl-L-valyloxy)-benzoate
(2.Og,
25 4.76 mmole) in dry acetone (30 ml) was added sodium iodide (3.15g, 21
mmole) and
the mixture was stirred overnight at room temperature. The mixture was
evaporated
under reduced pressure and extracted with ethyl actate/water. The organic
phase was
washed with a 5% sodium thiosulfate solution, dried with sodium sulfate and
evaporated under reduced pressure. Yield: 2.3g = 94%.
'H-NMR (CDC13) 1.02 (m, 6H) 2.38 (m, 1H) 4.56 (d, d , 1H) 5.14 (s , 2H) 5.30
(d,
1 H) 6.14 (s, 2H) 7.26-7.50 (m, 7H) 7.80(s, 1 H) 7.96 (d, 1 H)
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Example A-I-13
71
Iodomethvl3-lN-benzy_lox carbonyl-L-valyloxy)-,propionate
O O
I~O- v _O NHCBz
a) 3-buten-1-yl-3-(N-benzyloxycarbonyl)-propionate.
To a solution of 3-buten-1-of (2.16g, 30 mmole), N-benzyloxycarbonyl-1-valine
(8.29g, 33 mmole) and 4-dimethylaminopyridine (0.37g, 3 mmole) in
dichloromethane (80 ml) was added dicyclohexyl-carbodiimide (7.228, 35 mmole)
and the mixture was stirred overnight at room temperature. The mixture was
cooled
and the urethane was filtered. The solution was evaporated under reduced
to pressure and ethyl acetate (200 ml) was added. The organic phase was washed
twice
with 5% acetic acid, 5% sodium hydrogencarbonate and water. The organic phase
was dried with sodium sulfate and evaporated under reduced pressure. The
product
was isolated by silica gel column chromatography with hexane/ethyl acetate.
Yield:
8.3g = 90%.
is
b) 3-(N-benzyloxycarbonyl-L-valyloxy)-propanoic acid.
To a solution of 3-buten-1-yl -3-( N-benzyloxycarbonyl-L-valyloxy)-propionate
(9.2g, 30 mmole) in 150 ml benzene was added tetrabutylarnmonium bromide
2o (1.628, 5 mmole) and 100 ml water. The mixture was cooled to about
5°C and
potassium permanganate (14.82g, 90 mmole) was added in portions. The mixture
was stirred 2 hours at room temperature, diluted with water and decoiorized
by the addition of sodium bisulfate. The mixture was acidified with 2M
hydrogen
chloride and extracted 3 times with ethyl acetate. The combined organic phases
were
25 washed with water and dried with sodium sulfate. The solution was
evaporated under
reduced pressure and the product isolated by silica gel column chromatography
with
hexane/ethyl acetate. Yield: 5.4g = SS%.
'H-NMR (DMSO d-6) 0.90 (m, 6H) 2.5 (m, 2H) 3.88 (d, d, 1H) 4.32 (m, 2H) 5.03
30 (s, 2H) 7.36 (m, SH) 7.68 (d, 1H)
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c) Chloromethyl 3-(N-benzyloxycarbonyl-L-valyloxy)-propionate.
To a solution of 3-(N-benzyloxycarbonyl-L-valyloxy)propanoic acid (5.2g, 16.08
mmole) in 1,4-dioxane (50 ml) was added a 40% solution of tetrabutylammonium
hydroxide (10.43g, 16.08 mmole) and the mixture was stirred 2 hours at room
s temperature. The mixture was evaporated under reduced pressure and co-
evaporated
two times with 1,4-dioxane and two times with toluene. The dried product was
dissolved in 40 ml dichloromethane and chloroiodomethane (28.4g. 160 mmole)
was
added. The solution was stirred for two days at room temperature and
evaporated
under reduced pressure. Ethyl acetate (100 ml) was added and the organic phase
to washed twice with water, dried with sodium sulfate and evaporated under
reduced
pressure. The product was isolated by silica gel column chromatography. Yield:
2.2g
= 35%
d) Iodomethyl3-( N-benzyloxycarbonyl-L-valyloxy)-propionate.
1s To a solution of chloromethyl 3-(N-benzyloxycarbonyl-L-valyloxy)-propionate
(2.OSg, 5.51 mmole) in dry acetone (50 ml) was added sodium iodide (4.12g,
27.5
mmole) and the mixture was stirred overnight at room temperature. The mixture
was
evaporated under reduced pressure and extracted with ethyl acetate water. The
organic phase was washed with a 5% sodium thiosulfate solution, dried with
sodium
2o sulfate and evaporated under reduced pressure. Yield: 2.35g = 92%.
'H-NMR (CDC13) 0.94 (m, 6H) 2.17 (m, 1H) 2.68 (t, 2H) 4.40 (m, 3H) 5.12 (s,
2H)
5.91 (s, 2H) 7.26 (m, SH).
25 Example A-I-13A
1.3-bis(N tert-butoxvcarbonvl-L-valylox~r)-2~ropyl 1-iodoethyl carbonate
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a) 1,3-bis(N tent-butoxycarbonyl-1.-valyloxy)-2-propyl 1-chloroethyl
carbonate.
To a solution of 1,3-bis(N tent-butoxycarbonyl-L-valyloxy)-2-propanol (0.545
g,
1.11 mmol) in 5 mL dry CHZCl2 were added pyridine (540 pL, 6.68 mmol), with
cooling and stirring in an ice bath, followed by 1-chloroethyl chloroformate
(242 pL,
2.22 mmol). After 1 h, the reaction mixture was diluted with 5 mL CHZCIz and
washed with water (5 mL) and brine (5 mL). The organic phase was dried over
anhydrous NazS04 and concentrated on a rotavapor, coevapoating several times
with
toluene. Column chromatogaphy (silica, 4/1 petroleum ether - ethyl acetate)
gave the
1o chloride (596 mg, 90%) as a white solid.
b) 1,3-bis(N tert-butoxycarbonyl-L-valyloxy)-2-propyl 1-iodoethyl
carbonate.
A mixture of the chloride (596 mg, 1.0 mmol) from step (a) and NaI (684 mg,
4.57
~s mmol) in 10 ml dry MeCN was refluxed at 80 °C for 4 h. The reaction
mixture was
concentrated under vacuum and then partitioned between 30 mL diethyl ether and
10
mL water. The organic phase was washed with 5% aqueous sodium thiosulfate (2 x
5
mL), and the last aqueous layer was reextracted with ether (5 mL). The organic
phases were combined, washed with brine, dried over NazS04, and concentrated.
2o Flash column chromatography (silica, 4/1 petroleum ether - ethyl acetate)
gave a
fraction (275 mg) containing 80% iodide, as determined from 'H NMR, and small
amounts of the starting chloride and alkene from the elimination side
reaction.
'H NMR (250 MHz, CDCI3) 8 0.81-0.85 (m, 6H), 0.88-0.92 (m, 6H), 1.37 (s, 18H),
25 2.05 (m, 2H), 2.17 (d, 3H, J= 6.1 Hz), 4.12-4.46 (m, 6H), 5.00 (d,. 2H, J=
8.8 Hz),
5.11 (m, 1H), 6.68 and 6.69 (2 sets of q, 1H, J= 6.1 Hz).
L~,."
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Example A-I-14
74
3-(N benzvloxvcarbonvl-L-valvloxy)-2 2-dimethvlpropyl iod-omethvl carbonate
O O
I ~'O~O
HN O
O
(a) 3-(N benzyloxycarbonyl-L-valyloxy)-2,2-dimethyl-1-propanol.
A mixture ofN benzyloxycarbonyl-L-valine (2.50 g, 10.0 mmol), 2,2-dimethyl-1,3-
propanediol (5.30 g, 50.9 mmol), dicyclohexylcarbodiimide (2.60 g, 12.6 mmol),
and
4-dimethylaminopyridine (125 mg, 1.0 mmol) in 100 mL dry CHZCl2 was stirred
for
23 h. The reaction mixture was filtered and washed successively with 50 mL
each of
water, saturated aqueous NH4C1, saturated aqueous NaHC03, and water. The
organic
io phase was dried over anhydrous NazS04 and concentrated. The title compound
(2.99
g, 87%) was isolated by flash column chromatography (silica, 2/1 petroleum
ether -
ethyl acetate) as a white waxy solid.
(b) 3-(N benzyloxycarbonyl-L-valyloxy)-2,2-dimethylpropyl chloromethyl
carbonate.
Chloromethyl chloroformate (1.50 mL, 16.6 mmol) was added to a solution of the
alcohol (2.74 g, 8.12 mmol) from step (a) and pyridine (4.9 mL, 61 mmol) in 40
mL
dry CHZCIz, in an ice bath. After stirnng for 1 h, the mixture was diluted
with CHZC12
and washed successively with water, saturated NaHC03, and brine. The organic
2o phase was dried over anhydrous NazS04 and concentrated, coevaporating
several
times with toluene on a rotavapor. Flash column chromatography (silica, 2/1
petroleum ether - ethyl acetate) gave 3.31 g (95%) of the title compound.
(c) 3-(N benzyloxycarbonyl-L-valyloxy)-2,2-dimethylpropyl iodomethyl
carbonate.
A mixture of the chloride (3.14 g, 7.30 mmol) from step (b) and NaI (4.37 g,
29.2
mmol) in 73 mL dry MeCN was refluxed at 80 °C for 3 h. After removal of
solvent
under vacuum, the mixture was partitioned between 80 mL ethyl acetate and 40
mL
water. The organic phase was washed with 5% NazSz03, and then brine, dried
over
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anhydrous NazS04, and concentrated. Flash column chromatography (silica,
petroleum ether - ethyl acetate) gave 3.68 g (97%) of the title compound.
'H NMR (250 MHz, CDCI,) 8 0.88 and 0.96 (2d, 3H each), 0.98 (s, 6H), 2.18 (m,
IH), 3.94 and 4.02 (2s, 2H each), 4.32 (dd, 1H, J= 9.0, 4.7 Hz), 5.11 {s, 2H),
5.26 (d,
1H), 5.92 and 5.93 (ABq, 2H, JAB = S.lHz), 7.35 (s, SH).
Example A-I-15
1-(N benzvloxvcarbonvl-L-valvloxvl-2-methyl-2-nronvl iodomethvl carbonate
O O
I/'~O~O O .,,, N~O /
O H
io
(a) 1-(N benzyloxycarbonyl-L-valyloxy)-2-methyl-2-propanol
N Benzyloxycarbonyl-L-valine (2.02 g, 8.0 mmol), 4-dimethylaminopyridine (100
mg, 0.8 mmol), and ), and dicyclohexylcarbodiimide (2.04 g, 9.9 mmol, in 20 mL
CHZCIZ) were added to 2-methyl-1,2-propanediol (12.2 mmol) in 30 mL dry
CHZCI2,
1s with cooling in an ice bath. DMF (S mL) was added. After stirring for 5 h
at 10 °C ,
the reaction mixture was filtered, concentrated, and then redissolved in ethyl
acetate.
The organic solution was washed with saturated NaCI, dried over anhydrous
Na2S04,
and concentrated. Flash column chromatography (silica, 2/1 petroleum ether -
ethyl
acetate) gave 2.3 g of the title compound.
(b) 1-(N benzyloxycarbonyl-I,-valyloxy)-2-methyl-2-propyl chloromethyl
carbonate.
All of the alcohol from above was dissolved in 35 mL dry CHZCl2 and cooled in
an
ice bath. Pyridine (3.50 mL, 43.4 mmol) was added, followed by chloromethyl
chloroformate (1.30 mL, 14.4 mmol). After 1 h, the ice bath was removed and
stirring was continued for 2 h at ambient temperature.The mixture was diluted
with
CH,C12 (50 mL) and washed with water (50 mL), and then brine (2 x 25
mL).Drying
over anhydrous NazS04 of the combined organic phases and concentration under
vacuum, coevaporating several times with toluene, gave a yellow-brown oil that
was
3o subjected to flash column chromatography (silica, 2/1 petroleum ether -
ethyl
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acetate) to yield 2.86 g (86% from N benzyloxycarbonyl-L-valine) of the title
compound.
(c) 1-(N benzyloxycarbonyl-L-valyloxy)-2-methyl-2-propyl iodomethyl
carbonate.
A mixture of the chloride (2.84 g, 6.84 mmol) from step (b) and NaI (4.15 g,
27.2
mmol) in 68 mL dry acetonitrile was refluxed at 75 °C for 4 h. After
evaporation of
solvent under vacuum, the residue was partitioned between ethyl acetate (80
mL)
and water (40 mL), and the organic layer was washed with 5% NazSz03 (15 mL)
and
1o brine (25 mL). Drying the organic phase over anhydrous Na2S04 and
concentration
gave a yellow oil that was subjected to flash column chromatography (silica,
2/1
petroleum ether - ethyl acetate) to furnish 3.29 g (95%) of the title
compound.
'H NMR (250 MHz, CDCI,) 8 0.90 and 0.94 (2d, 3H each, J= 6.8 Hz), 1.52 (s,
6H),
2.17 (m, 1H), 4.35 (m, 1H), 4.22 and 4.39 (ABq, 2H, J,,B = 11.7 Hz), 5.10 (s,
2H},
5.30 (d, 1H), 5.86 (s, 2H), 7.34 (s, SH)
Example A-I-16
Iodomethvl 3.4-di-(N-CBZ-L-valvloxv)hvdrocinnamate
O O
CBz-NH O i O~I
CBz-N H
O
a) 4-Methoxybenzyl-3,4-dihydroxyhydrocinnamate.
3,4-Dihydroxycinnamic acid (6.5 g, 35.7 mmol) was dissolved in DMF (50 ml) and
cooled to 0°C on an ice-bath. 4-Potassium tert-butoxide (35.7 mmol),
was then added
and the mixture was left for approximately 30 min at 0°C, followed by
dropwise
adition of 4-methoxy-benzylchloride (39 mmol) in DMF (25 ml). The mixture was
allowed to reach room temperature and left over-night. The solvent was then
evaporated and the crude product was purified by chromatography (ethyl acetate-
hexane, 1:1) to give 6 g ofthe title compound (55%).
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b) 4-Methoxybenzyl-3,4-di-(N-CBZ-L-valyloxy)hydrocinnamate.
4-Methoxybenzyl-3,4-dihydroxyhydrocinnamate (5 g, 16.5 mmol), N,N
dimethylaminopyridine (2g, 16.5 mmol), N,N'- dicychlohexyl carbodiimide (8.5
g,
41.3 mmol) and Cbz-L-valine (10.4 g, 4I.3 mmol) were dissolved in
dichloromethane (50 ml). After 4 h, the the mixture was filtered and
evaporated onto
silica gel and purified by chromatography (hexane-EtOAc, 5:2 -a 3:2) to give
pure
title product (10.1 g, 79 %).
c) 3,4-Di-(N-CBZ-L-valyloxy)hydrocinnamic acid.
4-Methoxybenzyl-3,4-di-(N-CBZ-L-valyloxy)hydrocinnamate (10 g, 13 mmol) was
dissolved in dichloromethane and 1,1,1 trifluoroacetic acid (30 ml) and left
at
ambient temperature for 3.5 h. Evaporation under reduced pressure and
purification
by chromatography (chloroform-methanol, 10:1 ) yielded 6.7 g (80%) pure title
product.
'H NMR (CDC13 45 °C): 7. 24-7.0 (m, 13H), 5.65 (br s, 1H), 5.55 (hr s,
1H), 5.1 (m,
4H), 4.46 (m, 2H), 2.95 (t, 2H), 2.66 (t, 2H), 2.35 (m, 2H).
d) Chloromethyl 3,4-di-(N-CBZ-L-valyloxy)hydrocinnamate.
3,4-Di-(N-CBZ-L-valyloxy)hydrocinnamic acid (4.2 g, 6.47 mmol) was dissolved
in
2o dioxane (70 ml). Tetrabutylammonium hydroxide was added dropwise until
pH=8.
The solvent was then removed under reduced pressure The solid was redissolved
in
dioxane (30 ml) and toluene (30 ml) and evaporated. The procedure was repeated
twice (for removal of water). Dichloromethane (60 ml) and chloro-iodomethane
was
added in one portion and the mixture was left at ambient temperature for 6 h.
Evaporation of the solvent and purification by chromatography yielded 1.7 g
title
product (38%).
e) Iodomethyl 3,4-di-(N-CBZ-L-valyloxy)hydrocinnamate.
Chloromethyl 3,4-di-(N-CBZ-L-valyloxy)hydrocinnamate (1.9 g, 2.7 mmol) and
3o sodium iodide (2 g, 13.3 mmol) were dissolved in acetonitrile (50 ml) and
heated to
65° C for 60 min. The solvent was removed under reduced pressure and
the residue
was taken up in dichloromethane and filtrated. Removal of the solvent and
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purification by chromatography (ethyl acetate-hexane, 2:5) gave pure title
product
( 1.9 g, 90 %)
'H NMR (CDC13 45 °C): 7.34-7.02 (m, 13H), 5.89 {s, 2H), 5.64 (br s,
2H), 5.14-5.02
(m, 4H), 4.47 (m, 2H), 2.96 (t, 2H), 2.64 (t, 2H), 2.33 (m, 2H), 1.08-0.99 (m,
12H)
Example A-I-17
3-(N-CBZ-L-valyloxYlphenvl iodomethyl carbonate
O
O ~ O~O~I
CBz-NH
1o a) 3-(N-CBz-L-valyloxy)phenol.
CBz-L-valine (10 g, 40 mmol), 1,3-dihydroxybenzene (8.7g, 79 mmol)
N,N'dicychlohexylcarbodiimide (10.2g, 44 mmol) and 4-dimethylaminopyridine
(2.4
g, 20 mmol) were dissolved in DMF (50 ml) and left at ambient temperature
overnight. The reaction mixture was filtered, the solvent removed under
reduced
15 pressure and the crude product was taken up in dichloromethane and
filtered.
Removal of the solvent followed by purification by chromatography (chloroform-
methanol, 10:1) yielded pure title product (10.9 g, 79%).
b) (N-CBZ-L-valyloxy)phenyl chloromethyl carbonate.
20 3-(N-CBz-L-valyloxy)phenol (5.4 g, 15 7 mmol) was dissolved in
dichloromethane
(70 ml) and cooled in an ice-bath. Pyridine (1.2 g, 23.5 mmol was added
followed by
dropwise addition of 1-chloro-methylchloroformate (2.3 g, 18.8 mmol) in
dichloromethane (10 ml). The mixture was left at room temperature for 4 h.
Water
(25 ml) was then added and the phases were separated. The organic layer was
washed
25 with 0.01 M aqueous hydrochloric acid (25 ml). Purification by
chromatography
(ethyl acetate-hexane, 1:1 ) gave the title compound (4.5 g, 65 %)
c) 3-(N-CBZ-L-valyloxy)phenyl iodomethyl carbonate.
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(N-CBZ-L-valyloxy)phenyl chloromethyl carbonate (l.Sg, 3.44 mmol) and sodium
iodide (2 g, 13.3 mmol) were stirred at 60°C in acetonitrile (50 ml}
for 4.5 h.The
mixture was filered, the solvent removed and the crude product was taken up in
100
ml hexane-ethyl acetate, 1:1, and filtered through a sintered glass funnel,
packed with
2 cm silica gel. Removal of the solvent yielded pure title product (1.68 g,
92%)
'H NMR (CDCl3 45 °C): 7.38-7.02 (m, 9H), 6.03 (s, 2H), 5.2 (br s, 1H),
5.14 (s, 2H),
4.48 (m, 1 H), 2.3 0 (m, 1 H), 1.09-1.01 (m, 6H)
Example A-I-18
to Iodomethyl2-lN-CBZ-L-valvloxy}phenylacetate
O
O O~I
O
CBz-NH
a) 4-Methoxybenzyl 2-hydroxyphenylacetate.
2-hydroxyphenylacetic acid (10 g, 66 mmol) was dissolved in N,N dimethyl-
formamide (100 ml) and cooled on ice-bath. Potassium tert-butoxide (8.85 g, 78
15 mmoi) was added. The mixture was left for 30 min and allowed to reach room
temperature. 4-Methoxy-benzylchloride (11.7 g, 72 mmol) in N,N dimethyl-
formamide (30 ml) was then added dropwise, under nitrogen atmosphere and left
over-night. The solvent was evaporated under reduced pressure and the crude
mixture
was dissolved in ether (100 ml) and washed with water (25 ml), brine and dried
over
2o sodium sulphate. Chromatography (hexane-ethyl acetate, 2:1) followed by
recrystallization (hexane-ethyl acetate) gave the title compound (7.6 g, 42%).
b) 4-Methoxybenzyl2-(N-CBz-L-valyloxy)phenylacetate.
4-Methoxybenzyl 2-hydroxyphenylacetate 3g, 11 mmol), N,N,-dichyclohexyl-
25 carbodiimide (2.7 g, 13.2 mmol), dimethylaminopyridine (0.134 g, 1.1 mmol)
and
CBz-L-valine (3.3 g, 13.2 mmol) were dissolved in dichloromethane (50 ml).
After
the weekend the solid was filtered off, the solvent removed under reduced
pressure
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and the crude product purified by chromatography (ethyl aceta~,'ne~c~ne; ~ :2)
to give
the title compound (5.2 g, 93%).
c) 2-(N-CBz-L-valyloxy)phenylacetic acid.
5 4-Methoxybenzyl 2-(N-CBz-L-valyloxy)phenylacetate (4.25 g, 8.4 mmol), was
dissolved in dichloromethane (40 ml). Triflouroacetic acid (8 ml) was added
with
cooling on ice. The mixture was allowed to reach room temperature and stirred
for 40
min. The solvent was removed under reduced pressure and the crude product was
recrystallized twice (hexan-ethyl acetate + a small amount of dichloromethane)
to
1o give the title compound (2.6 g, 80 %).
'H NMR (CDCI, 45 °C): 7.35-7.08 (m, 9H), 5.35 (br s, 1H), 5.13 (s, 2H),
4.48
(m, 1H), 3.57 (s, 2H), 2.33 (m, 1H), 1.08 (d, 3H), 1.02 (d, 3H).
15 d) Chloromethyl2-(N-CBZ-L-valyloxy)phenylacetate.
This compound was prepared in poor yield from 2-(N-CBz-L-valyloxy)phenylacetic
acid (5.5 g, 14.3 mmol) by an unoptimized procedure essentially as described
in
Example A-I-16 d). Yield: 0.265 g
20 'H NMR (CDC13 45 °C): 7.28-7.01 (m, 9H), 5.55 (s, 2H), 5.2 (br s,
1H), 5.07 (s, 2H),
4.43 (m, 1H), 3.53 (s, 2H), 2.26 (m, 1H), 1.02 (d, 3H), 0.95 (d, 3H).
e) Iodomethyl 2-(N-CBZ-L-valyloxy)phenylacetate.
Chloromethyl 2-(N-CBZ-L-valyloxy)phenylacetate is treated with NaI and
purified
25 as described in the Examples above to yield the title compound.
Example A-I-19
Iodomethyl 4-lN-CBZ-L-valyloxyxy~phenvlacetate
O
O ~ O
CBz-NH
O~I
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io
a) 4-Methoxybenzyl 4-hydroxyphenylacetate.
Prepared from 4-hydroxyphenylacetic acid (10 g, 65.7 mmol) in 70 % yield by
the
same procedure as for Example A-I-18 a) above, but wherein the solvent for the
recrystallization was changed to hexane-ether.
b) 4-Methoxybenzyl4-(N-CBz-L-valyloxy)phenylacetate.
Prepared from 4-methoxybenzyl 4-hydroxyphenylacetate (3 g, 11 mmol) by the
same
procedure as for Example A-I-18 b) in 87 % yield. Solvent for chromatography:
ethyl
acetate-hexane, 1:2.
c) 4-(N-CBZ-L-valyloxy)phenylacetic acid.
Prepared in 82 % yield from 4-methoxybenzyl 4-(N-CBz-L-valyloxy)phenylacetate
(1.6 g, 288 mmol) by the procedure described for Example A-I-18 c). Solvent
for
recrystallization: hexane-ether and a small amount of dichloromethane.
'H NMR (CDC13 45 °C): 7.36-7.27 (m, 7H), 7.02 (d, 2H), 5.25 (d, 1H),
5.14 (s, 2H),
4.52 (m, 1H), 3.64 (s, 2H), 2.3 (m, 1H), 1.08 (d, 3H), 1.02 (d, 3H).
d) Chloromethyl4-(N-CBZ-L-valyloxy)phenylacetate.
Prepared from 4-(N-CBZ-L-valyloxy)phenylacetic acid (3 g, 7.8 mmol) in 26
2o yield by the same procedure as described for Example A-I-18 d). Solvent for
chromatography: hexane-ether, 3:2.
e) Iodomethyl 4-(N-CBZ-L-valyloxy)phenylacetate.
Chloromethyl 4-(N-CBZ-L-valyloxy)phenylacetate (0.83 g, 1.9 mmol) and sodium
iodide (1.15 g, 7.6 mmol) were heated in acetonitril (45 ml) for 5 h. The
mixture was
filtrated, the solvent removed, taken up in dichloromethane and filtrated
again.
Evaporation and purification by chromatography (ether-hexane, 2:3) yielded the
title
product (0.8 g, 80 %).
'H NMR (CDCI, 45 °C): 7.38-7.09 (m, 4H), 5.$4 (s, 1H), 5.30 (br s, 1H),
5.15
(s, 2H), 4.5 (m, 1H), 3.56 (s, 2H), 2.36 (m, 1H), 1.10 (d, 3H), 1.00 (d, 3H).
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Example A-I-20
Iodometh~l 4-(2-N-benzylox~rcarbonyl-L-val ,~loxyethyl benzoate
a) 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl~toluene.
To a cooled solution of 4-methylphenylethanol-2 (S.Og, 36.7 mmole), 4
dimethylaminopyridine (0.98g, 8 mmole) and N-benzyloxycarbonyl-L-valine
( 10.05g, 40 mmole) in dichloromethane ( 120 ml) was added dicyclohexyl-
carbodiimide (9.1 g, 44 mmole) and the mixture was stirred overnight at room
temperature. The mixture was cooled and the urethane was filtered. The
solution was
1o evaporated under reduced pressure and ethyl acetate (250 ml) was added. The
organic
phase was washed twice with 5% acetic acid, 5% sodium hydrogencarbonate and
water. The organic phase was dried with sodium sulfate and evaporated under
reduced pressure. The product was isolated by silica gel column chromatography
with toluene/acetone. Yield: 13.3g = 97%
b) 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl)- benzoic acid.
To a cooled mixture of chromic anhydride (7.558, 75 mmole) in acetic acid (100
ml)
was added dropwise a solution of 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl)-
toluene (9.3g, 25.1 mmole) in acetone (50 ml). The mixture was stirred at room
temperature for 3 days and reduced to about 100 ml. 600m1 10% sodium chloride
2o solution was added and the mixture was extracted four times with ethyl
acetate. The
organic phase was washed with brine and dried with sodium sulfate. The
solution
was evaporated under reduced pressure and the product was islolated by silica
gel
column chromatography with dichloromethane/methanol. Yield : 2,1 g = 21 %.
'H-NMR (CDC13) 0.79 (d, 3H) 0.90 (d, 3H) 2.08 (m,lH) 3.04 (t, 2H) 4.28 (d,
d,lH)
4.39 (m, 2H) 5.11 (s, 2H) 5.26 (d, 1H) 7.34 (m, 7H) 8.04 (d, 2H)
c) Chloromethyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl)benzoate.
To a solution of 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl)benzoic acid (2.Og,
5.0
3o mmole) in 1,4-dioxane (20 ml)was added a 40% solution of tetrabutylammonium
hydroxide (3.1 g, 4.75 mmole) and the mixture was stirred 2 hours at room
temperature. The mixture was evaporated under reduced pressure and
coevaporated
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two times with 1,4-dioxane and two times with toluene. The dried product was
dissolved in dichloromethane (10 ml) and iodochloromethane (I3.2g, 75 mmole)
was
added The solution was stirred overnight at room temperature and evaporated
under
reduced pressure. About 50 ml ethyl acetate were added and the organic phase
washed twice with water, dried with sodium sulfate and evaporated under
reduced
pressure. The product was isolated by silica gel column chromatography.Yield:
O.Sg
=23%
d) Iodomethyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl) benzoate.
o To a solution of chloromethyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethyl)
benzoate
(O.Sg, 1.11 mmole). In dry acetone (10 ml) was added sodium iodide (0.75g, 5.0
mmole) and the mixture was stirred overnight at room temperature. The mixture
was
evaporated under reduced pressure and extracted with ethyl actate/water. The
organic
phase was washed with a 5% sodium thiosulfate solution, dried with sodium
sulfate
is and evaporated under reduced pressure.Yield: 0.53g = 88%.
'H-NMR (CDCI,) 0.88 (d, 3H) 0.90 (d, 3H) 2.08 (m, 1H) 3.02 (t, 2H) 4.28 (d, d,
1H)
4.38 (m, 2H) 5.10 (s, 2H) 5.22 (d, 1H) 6.15 (s, 2H) 7.35(m, 7H) 7.98 (d, 2H )
20 Example A-I-21
Iodometh,~(N-benzvloxycarbonyl-L-isoleucvloxvmethvll
2-meth~,rl,propionate.
a) 4-methoxybenzyl 2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl)-
2s 2-methyl propionate.
To a cooled solution of 4-methoxybenzyl 2-(hydroxymethyl)-2-methyl propionate
(6.Og, 25 mmole), 4-dimethylaminopyridine (0.61 g, 5 mmole) and N-
benzyloxycarbonyl-L-isoleucine (6.90g, 26 mmole) in dichloromethane (100 ml)
was
added dicyclohexyl-carbodiimide (6.2g, 30 mmole) and the mixture was stirred
3o overnight at room temperature.The mixture was cooled and the urethane was
filtered.
The solution was evaporated and 200 ml ethyl acetate was added, The organic
phase
was washed twice with S% acetic acid, 5% sodium hydrogencarbonate and water.
The organic phase was dried with sodium sulfate and evaporated under reduced
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pressure. The product was isolated by silica gel column chromatography with
toluene/acetone.Yield: 11.7g = 96%.
2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl)-2-methyl) propionic acid.
To a solution of 4-methoxybenzyl 2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl}-
2-methyl propionate (ll.Og, 22.6 mmole) in 100 ml dichloromethane was added
trifluoroacetic acid (15 ml) and the mixture was stirred overnight at room
temperature. The solution was evaporated under reduced pressure and
coevaporated
two times with toluene. The residue was stirred 1 hour with 100 ml ethanol and
the
1o white solid was filtered (byproduct). The solution was evaporated under
reduced
pressure and the product was isolated by silica gel column chromatography with
hexane/ethyl acetate. Yield: 7.4g = 89%.
'H-NMR (CDCI,) 0.90 (m, 6H) 1.26 (m, 8H) 1.88 (m, 1H) 4.12 (d, d, 2H) 4.38
(d, d, 1H) 5.10 (s, 2H) 5.32 (d, 1H) 7.28 (m, 5H)
c) Chloromethyl 2-(N-benzyloxycarbonyl-L-isoleucyloxy)-2-methyl
propionate.
To a solution of 2-(N-benzyloxycarbony-L-isoleucyloxymethyl)-2-methyl
2o propionic acid (7.Og, 19 mmole) in 80 ml 1,4-dioxane was added a 40%
solution of
tetrabutylammonium hydroxide (12.4g, 19 mmole) and the mixture was stirred 2
hours at room temperature. The mixture was evaporated under reduced pressure
and
co-evaporated two times with 1,4-dioxane and two times with toluene. The dried
product was dissolved in 25 ml dichloromethane and iodochloromethane (33.7g,
190
mmole) was added . The solution was stirred overnight at room temperature and
evaporated under reduced pressure. About 100 ml ethyl actate was added and the
organic phase washed twice with water, dried with sodium sulfate and
evaporated under reduced pressure. The product was isolated by silica gel
column
chromatography with toluene/acetone.Yield: 4.2 = 54%
d) Iodomethyl2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl)-2-methyl
propionate.
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To a solution of chloromethyl 2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl)-2-
methyl propionate (3.Og, 7.2 mmole) in 50 ml dry acetone was added sodium
iodide
(4.8g, 32 mmole) and the mixture was stirred overnight at room temperature.
The
mixture was evaporated under reduced pressure and extracted with ethyl actate
5 water. The organic phase was washed with a 5% sodium thiosulfate solution,
dried
with sodium sulfate and evaporated under reduced pressure. Yield: 3.3g = 90%.
'H-NMR (CDC13) 0.93 (m, 6H) 1.23 (m, 8H) 4.12 (m, 2H) 4.38 {d, d, 1H) 5.10
(s, 2H) 5.26 (d, 1H) 5.92 (m, 2H) 5.35 (m, SH)
Example A-I-22
Iodometh 1~4- N-benzylox cy arbonyl-L-va~loxy)cvclohexanoate.
a) 4-Methoxybenzyl 4-hydroxycyclohexanoate.
To a solution of ethyl 4-hydroxycyclohexanoate (8.61g, 50 mmole) in 50 ml
ethanol
was added a solution of potassium hydroxide 85% (3.63g, 55 mmole) and the
mixture was stirred for 6 hours at 70°C. The mixture was evaporated
under reduced
pressure, coevaporated two times with N,N-dimethylformamide and reduced to
about
100 ml. 4-Methoxybenzyl chloride (9.4g, 60 mmole) was added and the mixture
was
2o stirred for 18 hours at 60°C. The mixture was evaporated under
reduced pressure and
250 ml ethyl acetate was added. The organic phase was washed five times with
water, dried with sodiun sulfate and evaporated under reduced pressure. Yield:
13.2g
=100% (crude)
b) 4-methoxybenzyl4-(N-benzyloxycarbonyl-L-valyloxy)-
cyclohexanoate.
To a cooled solution of 4-methoxybenzyl 4-hydroxycyclohexanoate (7.Sg, 28
mmole), 4-dimethylaminopyridine (0.73g, 6 mmole) and N-benzyloxycarbonyl-L-
valine (7.54g, 30 mmole) in dichloromethane (90 ml) was added dicylohexyl-
3o carbodiimide (6.8g, 33 mmole) and the mixture was stirred for 2 days at
room
temperature. The mixture was cooled and the urethane was filtered. The
solution was
evaporated and 250 ml ethyl acetate was added.The organic phase was washed
twice
with 5% acetic acid, 5% sodium hydrogencarbonate and water. The organic phase
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was dried with sodium sulfate and evaporated under reduced pressure. The
product
was isolated by silica gel column chromatography with toluene/acetone.Yield :
13g
= 93%
c) 4-(N-benzyloxycarbonyl-L-valyloxy) cyclohexanoic acid.
To a solution of 4-methoxybenzyl 4-(N-benzyloxycarbonyl-L-valyloxy)-
cyclohexanoate (12g, 24.1 mmole} in dichloromethane (100 ml) was added
trifluoroacetic acid (20 ml) and the mixture was stirred for 3 hours at room
temperature. The solution was evaporated under reduced pressure and
coevaporated
1o two times with toluene. The residue was stirred 1 hour with about 100 ml
ethanol and
the white solid was filtered (byproduct). The solution was evaporated under
reduced
pressure and the product was isolated by silica gel column chromatography with
toluene/acetone. Yield: 6.8g = 74%.
'H-NMR (CDC13) 0.91 (m, 6H) 1.52-2.54 (m, lOH) 4.28 (m, 1H) 4.82-5.08 (m, 1H)
5.11 (s, 2H) 5.28 (d, 1 H) 7.36 (m, SH)
d) Chloromethyl4-(N-benzyloxycarbonyl-L-valyloxy)-cyclohexanoate.
To a solution of 4-(N-benzyloxycarbonyl-L-valyloxy) cyclohexanoic acid (6.6g,
20
2o mmole) in 1,4-dioxane (70 ml) was added a 40% solution of
tetrabutylammonium
hydroxide {11.34g, I7.5 mmole) and the mixture was stirred 2 hours at room
temperature. The mixture was evaporated under reduced pressure and co-
evaporated
two times with 1,4-dioxane and two times with toluene. The dried product was
dissolved in 60 ml dichloromethane and iodochloromethane (30.98, 175 mmole)
was
added. The solution was stirred for two days at room temperature and
evaporated
under reduced pressure. About 100 ml ethyl actate was added and the organic
phase
washed twice with water, dried with sodium sulfate and evaporated under
reduced
pressure. The product was isolated by silica gel column chromatography with
toluene/acetone. Yield: 4.1g = 55%.
e) Iodomethyl 4-(N-benzyloxycarbonyl-L-valyloxy)-cyclohexanoate.
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To a solution of chloromethyl 4-{N-benzyloxycarbonyl-L-valyloxy)-
cyclohexanoate
(4.Og, 9.4 mmole) in dry acetone (50 ml) was added sodium iodide (6.3g, 42
mmole)
and the mixture was stirred overnight at room temperature. The mixture was
evaporated under reduced pressure and extracted with ethyl actate water. The
organic phase was washed with a 5% sodium thiosulfate solution, dried
with sodium sulfate and evaporated under reduced pressure.Yield 4.Sg = 93%.
'H-NMR (CDC13) 0.90 (m, 6H) 1.52-2.02 (m, 8H) 2.18 (m, 1H) 2.43 {m, 1H) 4.30
(m, 1H) 4.76-5.08 (m, 1H) 5.11 (s, 2H) 5.26 (d, 1H) 5.91 (d, 2H) 7.34 (m, SH)
to
Example A-I-23
Iodomethyl 2-(N-benzyloxycarbonyl-L-valyloxymeth~)-2-ethyl but~rrate
O
O~I
N-CBz-Valy1-'O
a) 2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-ethylbutan-1-ol.
To a cooled solution of 2-ethyl-2-hydroxymethyl-butan-1-of (33.1 g, 250
mmole), 4
dimethylaminopyridine ( 1.22g, 10 mmole) and N-benzyloxycarbonyl-L-valine
(12.6g, 50 mmole) in 350 ml dichloromethane was added dropwise a solution of
dicyclohexyl-carbodiimide ( 12.4g, 60 mmole) in 50 ml dichloromethane. The
mixture was stirred 2 days at room temperature and cooled. The urethane was
filtered
2o and the solution evaporated under reduced pressure. 350 ml ethyl
acetate was added and the organic phase was washed twice with 5% acetic acid,
5%
sodium-hydrogencarbonate and water. The organic phase was dried with sodium
sulfat and evaporated under reduced pressure. The product was isolated by
silica gel
column chromatography with dichloromethane/methanol. Yield: 16.4g = 90%.
c) 2-(N-benzyloxycarbonyl-L-valyloxymethyl )-2-ethyl-butyric acid.
To a cooled mixture of chromic anhydride (B.Sg, 85,2 mmole) in 100 ml acetic
acid
was added dropwise a solution of 2-(N-benzyloxycarbonyl-L-valyoxymethyl)-2-
ethyl-butan-1-of (10.4g, 28.4 mmole) in 50 ml acetone and the mixture was
stirred 24
3o hours at room temperature. The mixture was added to 1000 ml 10% sodium
chloride
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solution and extracted four times with ethyl acetate. The organic phase was
washed
twice with brine, dried with sodium sulfate and evaporated under reduced
pressure.
The product was isolated by silica gel column chromatography with hexane/ethyl
acetate. Yield: 7g = 65%.
'H-NMR (CDCI,) 0.88 (m, 12H) 1.67 (m, 4H) 2.14 (m, 1H) 4.26 (m, 3H) 5.10
(s, 2H) 5.30 (d, 2H) 7.34 (m, SH)
d) Chloromethyl 2-(N-benzyoxycarbonyl-L-valyloxymethyl -2-ethyl
1o butyrate.
To a solution of 2-(N-benzyloxycarbony-L-valyloxymethyl~2-ethyl-butyric acid
(7.2g,18,9 mmole) in 1,4-dioxane (80 ml) was added a 40% solution of
tetrabutylammonium hydroxide (12.26g, 18.9 mmole) and the mixture was stirred
2
hours at room temperature. The mixture was evaporated under reduced pressure
and
co-evaporated once with 1,4-dioxane and two times with toluene. The dried
product
was dissolved in 30 ml dichloromethane and iodochloromethane (49.48, 280
mmole)
was added. The solution was stirred for two days at room temperature and
evaporated
under reduced pressure. About 100 ml ethyl actate were added and the organic
phase
washed twice with water, dried with sodium sulfate and evaporated under
reduced
2o pressure. The product was isolated by silica gel column chromatography.
Yield: 5.2g
= 63%.
e) Iodomethyl 2-( N-benzyloxycarbonyl-L-valyloxymethyl)-2-ethyl
butyrate.
To a solution of chloromethyl 2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-ethyl
butyrate (S.Og, 11.7 mmole) in dry acetone (60 ml) was added sodium iodide
(7.Sg,
50 mmole) and the mixture was stirred overnight at room temperature. The
mixture
was evaporated under reduced pressure and extracted with ethyl actate water.
The
organic phase was washed with a 5% sodium thiosulfate solution, dried with
sodium
3o sulfate and evaporated under reduced pressure. Yield: 5.4g = 90%.
'H-NMR (CDC13) 0.92 (m, 12H) 1.65 (m, 4H) 2.18 (m, 1H) 4.28 (m, 3H) 5.10
(s, 2H) 5.22 (d, 1 H) 5.92 (s, 2H) 7.36 (m, SH)
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Example A-I-24
2JN-(,iodomethoxycarbonyl -~aminol-2-meth 1-y 1-(N-benzyloxvcarbon
val~yLpropane
O
o NJL.o~i
CBzN H o H
a) 2-(N-tert.-butyloxycarbonylamino)-2-methyl-1-(N-benzyloxycarbonyl-
L-valyloxy}-propane.
To a cooled solution of 2-(N-(tert.-butyloxycarbonyl)-amino)-2-methylpropan-1-
oI
(J. Am. Chem. Soc 113 (1991) p 8883) (4.73g, 25 mmole), 4-dirnethylamino-
to pyridine (0.61g, 5 mmole) and N-benzyloxycarbonyl-L-valine (6.28g, 25
mmole) in
dichloromethane (70 ml) was added dicyclohexyl-carbodiimide (6.19g, 30 mmole)
and the mixture was stirred 2 days at roommtemperature. The mixture was
cooled,
the urethane was filtered and the solution evaporated under reduced pressure.
Ethyl
acetate (200 ml) was added and the organic phase was washed twice with 5%
acetic
1s acid, 5% sodium hydrogencarbonate and water. The organic phase was dried
with,sodium sulfate and evaporated under reduced pressure. The product was
isolated
by silica gel column chromatography with hexane/ethyl acetate.Yield: 10.28 =
96%.
b) 2-amino-2-methyl-1-(N-benzyloxycarbonyl-L-valyloxy)-propane.
2o To a solution of 2-(N-(tert.-butyloxycarbonyl)-amino-2-methyl-1-(N-
benzyloxycarbonyl-L-valyloxy)-propane (lOg, 23 mmole) in dichloromethane (150
ml) was added trifluoroacetic acid (30 ml) and the mixture was stirred for 1
hour at
room temperature. The solution was evaporated under reduced pressure and 10%
sodium carbonate solution was added. The product was extracted four times with
2s dichloromethane, dried with sodium sulfate and evaporated under reduced
pressure.
The product was isolated by silica gel column chromatography with
dichloromethane/methanol. Yield: 3.Og = 40% (crude)
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c) 2-(N-(chloromethoxycarbonyl)-amino)-2-methyl-1- (N-
benzyloxycarbonyl-L-valyloxy)-propane.
To a solution of 2-amino-2-methyl-1-(N-benzyloxycarbonyl-L-valyloxy)-propane
(2.9g, 9 mmole) and pyridine (2 ml) in dichloromethane (50 ml) was added
5 chloromethyl chloroformate(I.SSg, 12 mmole) and the mixture was stirred for
3
hours at room temperature. The mixture was evaporated under reduced pressure
and
ethyl acetate was added. The organic phase was washed with water, dried with
sodium sulfate and evaporated under reduced pressure. The product was isolated
by
silica gel column chromatography with hexane/ethyl acetate.Yield: l.lg = 29%.
d) 2-(N-(iodomethoxycarbonyl)-amino)-2-methyl-1-(N-
benzyloxycarbonyl-L-valyloxy)-propane.
To a solution of 2-(N-(chloromethoxycarbonyl)-amino)-2-methyl-1-(N-
benzyloxycarbonyl-L-valyloxy)propane (I.OSg, 2.53 mmole) in dry acetone (20
ml)
was added sodium iodide (1.8g, 12 mmole) and the mixture was stirred for 36
hours
at room temperature. The mixture was evaporated under reduced pressure and
ethyl acetate and water were added. The organic phase was washed with 10%
sodium
thiosulfate solution and water. The organic phase was dried with sodium
sulfate and
evaporated under reduced pressure.Yield: 1.04g = 81%.
'H-NMR (CDCI,) 0.92 (m, 6H) 1.35 (s, 6H) 2.10 (m,lH) 3.88 (m, 1H) 4.35
(m, 2H) 5.11 (s, 2H) 5.32 (d, 1H) 5.82 (s, 1H) 5.91 (s, 2H) 7.35 (m, SH)
Example A-I-25
1-(2-N-CBz-L-valvlox~~)-6-oxo-1,6-dihydro~vridine-3-carboxylic acid
iodometh- Iy ester
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O
O
O~N~
O-Cbz-Val
a) 6-oxo-1,6-dihydro-pyridine-3-carboxylic acid 4-methoxybenzyl ester.
To a solution of 6-hydroxynicotinic acid (4.87 g, 35 mmol) in DMF (100 mL) at
room temperature, was added potassium tert-butoxide (3.93 g, 35 mmol). The
reaction mixture was stirred at 60 °C for lh. 4-Methoxybenzylchloride
(8.30 g, 53
mmol) was added and the reaction mixture was stirred at 60 °C for 4h.
The DMF was
evaporated under vacuum, the residue was dissolved in ether (200 mL) and
washed
with water (3 x 100 mL). The organic phase was dried with NazS04 and
evaporated
to give 4.41 g of 6-oxo-1,6-dihydro-pyridine-3-carboxylic acid 4-methoxybenzyl
1 o ester.
b) 1-(2-Hydroxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic acid 4-
methoxybenzyl ester.
To a solution of 6-oxo-1,6-dihydro-pyridine-3-carboxylic acid 4-methoxybenzyl
is ester (4.41 g, 17 moral) and KzC03 (2.58 g, 18.7 mmol) in DMF (100 mL) at
room
temperature, was added 2-bromoethanol (2.02 g, 16.2 mmol). The reaction
mixture
was stirred at 80 °C for 30h, whereupon the DMF was evaporated under
vacuum.
The crude product was column chromatographed (silica gel, 2-~S% MeOH in
CHzCl2), to give 3.91 g of 1-(2-hydroxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
2o carboxylic acid 4-methoxybenzyl ester.
c) 1-(2-N-CBz-L-valyloxyethyl )-6-oxo-1,6-dihydro-pyridine-3-carboxylic
acid 4-methoxybenzyl ester.
To a mixture of DCC (5.06 g, 24.5 mmol), DMAP (318 mg, 2.6 mmol) and N-CBz-
2s L-valine (6.48 g, 25.8 mmol) in CHzCIz (200 mL) at 0 °C, was added
dropwise a
solution of 1-(2-hydroxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic acid 4
methoxybenzyl ester (6.40 g, 24 mmol) in CHZCl2 (200 mL). After lh at 0
°C, the
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temperature of the reaction mixture was allowed to assume room temperature and
then the mixture was stirred for Sh at room temperature. The mixture was
filtered
through a glass filter and the solvent was removed under reduced pressure. The
crude
product was column chromatographed (silica gel, 2-~5% MeOH in CHZC12), to give
6.81 g 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic
acid
4-methoxybenzyl ester.
d) 1-(2-N-CBz-L-valyloxyethyl)-2-pyridone-5-carboxylic acid.
To a solution of 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
to carboxylic acid 4-methoxybenzyl ester (6.46 g, 12 mmol) in CHzCl2 (85 mL)
at room
temperature, was added trifluoroacetic acid (15 mL). After lh at room
temperature,
the reaction mixture was concentrated under reduced pressure. The crude
product
was column chromatographed (silica gel, 3-~6% MeOH in CHZCIz), to give 4.91 g
1-
(2-N-CBz-L-valyloxyethyl)-2-pyridone-5-carboxylic acid.
i5
'H-NMR (CDC13): 12.15 (br s, 1H), 8.29 (d, J= 2.2 Hz, 1H), 7.93 (dd, J= 9.5,
2.2
Hz, 1 H), 7.31 (m, SH), 6.69 (d, J = 9.5 Hz, 1 H), 5.53 (d, 1 H), 5.07 (s,
2H), 4.52-
4.05 (m, SH), 2.20-2.00 (m, 1H), 0.90 (d, 3H), 0.81 (d, 3H).
2o e) 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic
acid chloromethyl ester.
To a solution of 1-(2-N-CBz-L-valyloxyethyl)-2-pyridone-S-carboxylic acid
(4.91 g,
11.8 mmol) in dioxane (200 mL), was added dropwise a 40% aqueous solution of
tetrabutylammonium hydroxide (7.65 g). After stirring for 5 min, the solution
was
25 evaporated to dryness through co-evaporation with dioxane and toluene. The
residue
was dissolved in dichloromethane (200 mL) and then chloroiodomethane (8.74 mL,
120 mmol) was added and the solution was stirred for 12h at room temperature:
The
solution was concentrated under reduced pressure and the residue was shaken
with
hexane / ethyl acetate (1:1 v/v, 200 mL). The yellow crystalline solid was
filtered off
30 and the filtrate was washed with aqueous solution of sodium thiosulfate
(0.1 M) and
the filtered through anhydrous sodium sulfate and evaporated to dryness. The
residue
was column chromatographed (silica gel, 2-4% MeOH in CH,CIz), to give 1.80 g
of
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1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic acid
chloromethyl ester.
f) 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic
acid iodomethyl ester
To a solution of 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
carboxylic acid chloromethyl ester (1.80 g, 3.87 mrnol) in acetonitrile (30
mL), was
added sodium iodide (2.32 g, 15.5 mmol). The solution was stirred for 4 h at
60 °C.
The resulting suspension was filtered and the filtrate was evaporated. The
residue
to was dissolved in CHZC12 and washed with aqueous sodium thiosulfate (0.1 M).
The
organic phase was dried (Na2S04) and concentrated under reduced pressure. The
crude product was column chromatographed (silica gel, 1% MeOH in CHZC12), to
give 2.04 g 1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
carboxylic
acid iodomethyl ester.
'H-NMR (CDC13): 8.19 (d, J= 2.5 Hz, 1H), 7.79 (dd, J= 9.6, 2.5 Hz, 1H), 7.32
(m,
SH), 6.52 (d, J = 9.6 Hz, 1 H), 6.04 (s, 2H), 5.3 8 (d, 1 H), 5.07 (s, 2H),
4.54-4.06
(m, SH), 2.20-2.00 (m, 1H), 0.91 (d, 3H), 0.81 (d, 3H).
2o Example A-I-26
Iodomethvl 5-ffN benzylox carbonyl-L-valvloxvlmethyl]-2-furoate
O
\ !
O- 'NH O--~
/ O
O O
O
(a) 5-[(N Benzyloxycarbonyl-L-valyloxy)methyl]-2-furaldehyde.
A solution of 5-(hydroxymethyl)-2-furaldehyde (1.00 g, 7.69 mmol) in 5 mL dry
CHZC12 was added to a mixture of N benzyloxycarbonyl-L-valine (2.40 g, 9.57
mmol), N,N'-dicyclohexylcarbodiimide (2.00 g, 9.69 mmol), and 4-dimethyl-
aminopyridine ( 117 mg, 0.96 mmol) in 45 mL CHzCIz. After stirnng overnight,
the
reaction slurry was filtered, concentrated under vacuum, and subjected to
flash
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column chromatography (silica, 2/1 petroleum ether - ethyl acetate to give the
valine
ester (quantitative yield).
(b) 5-[(N Benzyloxycarbonyl-L-valyloxy)methyl]-2-furoic acid
A solution of NaClOz (2.8 mmol) in 3 mL water was added dropwise to a stirred
solution of 5-[(N benzyloxycarbonyl-L-valyloxy)methylJ-2-furaldehyde (798 mg,
2.22 mmol) from step (a) in 3 mL MeCN, with cooling in an ice bath. After 2.5
h, the
ice bath was removed, 2 mL more MeCN was added, and the two-phase liquid
reaction mixture was stirred at room temperature for 25 h. The reaction
mixture was
io diluted with water, made basic with saturated NaHC03, and extracted with
ethyl
acetate (3 x SO mL). The separated aqueous solution was acidified to pH 2 with
5%
aqueous HCl and extracted with ethyl acetate (3 x 50 mL). This second ethyl
acetate
solution was washed with brine, dried over anhydrous Na2S04, and evaporated to
dryness under vacuum to give the carboxylic acid (287 mg, 34%) which was used
in
the next step without further purification.
(c) Chloromethyl 5-[(N benzyloxycarbonyl-L-valyloxy)methylj-2-furoate.
Tetrabutylammonium hydroxide (40 wt. % solution in water, 0.55 mL, 0.84 mmol)
was added to the carboxylic acid (286 mg, 0.76 mmol) from step (b) in 5 mL
2o dioxane. The yellow solution was concentrated under vacuum, coevaporating
several
times with dioxane, toluene, and, lastly, CHZC12. The residue was charged with
10
mL dry CHZCIz and chloroiodomethane (0.55 mL, 7.55 mmol) was added. After
stirring for 20.5 h, the reaction mixture was concentrated and subjected to
flash
column chromatography (silica, 2/1 petroleum ether - ethyl acetate) to give
the
chloromethyl ester (137 mg, 42%).
(d) Iodomethyl 5-[(N benzyloxycarbonyl-L-valyloxy)methyl]-2-furoate.
All of the chloromethyl ester (137 mg, 0.32 mmol) from step (c) was refluxed
with
NaI (195 mg, 1.3 mmol) in 3.2 mL dry MeCN at 70 °C for 4 h. The
solvent was
3o removed under vacuum and the residue was subjected to flash column
chromatography (silica, 3/1 petroleum ether - ethyl acetate) to give the
iodomethyl
ester (152 mg, 92%).
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'H NMR (250 MHz, CDC13) 8 0.84 and 0.93 (2d, 3H each, J= 6.8 Hz), 2.16 (rn,
1H),
4.33 (dd, 1H, J= 9.1, 4.7 Hz), 5.09-5.21 (m, 4H), 5.36 (d, 1H, J= 9.1 Hz),
6.08 (s,
2H), 6.52 (d, 1H, J= 3.4 Hz), 7.19 (d, 1H, J= 3.5 Hz), 7.33 (s, SH).
Example A-I-27
Iodomethyl 4-(2-N-benzylox can 1-~ L-valyloxyethoxyl benzoate
a) 4-Methoxybenzyl 4-(2-hydroxyethoxy)benzoate.
To a solution of 4-methoxybenzyl 4-hydroxybenzoate (7.Og, 27 mmole) in dry N,N-
io dimethylformamide (50 ml) was added potassium carbonate (4.15g, 30 mmole)
and
2-bromoethanol.The mixture was stirred 48 hours at 80°C, evaporated
under reduced
pressure and ethyl acetate and water were added. The organic phase was washed
five
times with water and dried with sodium sulfate. The solution was evaporated
under
reduced pressure and the product was isolated by silica gel column
chromatography
15 with hexane/ethyl acetate.Yield: 6.8g = 83%.
b) 4-methoxybenzyl4-(2-N-benzyloxycarbonyl-L-
valyloxyethoxy)benzoate.
To a solution of 4-methoxybenzyl 4-(2-hydroxyethoxy) benzoate (6.6g, 21.8
mmole), 4-dimethylaminopyridine (0.61g, 5 mmole) and N-benzyloxycarbonyl-L-
2o valine (6.3g, 25 mmole) in dichloromethane (80 ml) was added dicyclohexyl-
carbodiimide (5.2g, 25 mmole) and the mixture was stirred overnight at room
temperature. The mixture was cooled and the urethane was filtered.The solution
was
evaporated and ethyl acetate (200 ml) was added. The organic phase was washed
twice with 5% acetic acid, 5% sodium hydrogencarbonate and water. The organic
25 phase was dried with sodium sulfate and evaporated under reduced pressure.
The
product was isolated by silica gel column chromatography with
dichloromethane/methanol. Yield: 10.6g = 90 %.
c) 4-(2-N-benzyloxycarbonyl-L-valyloxyethoxy)-benzoic acid.
30 To a solution of 4-methoxybenzyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethoxy)
benzoate ( 10.2g, 19.04 mmole) in dichloromethane ( 100 ml) was added
trifluoroacetic acid (20 ml) and the mixture was stirred 3 hours at room
temperature.
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The solution was evaporated under reduced pressure and co-evaporated two times
with toluene. The product was isolated by silica gel column chromatography.
Yield:
6.9g = 87%.
'H-NMR (CDCl3) 0.94 (m, 6H) 2.18 (m, 1H) 4.22- 4.68 (m, SH) 5.10 (s, 2H)
6.94 (d, 2H) 7.35 (m, SH) 8.05 (d, 2H)
d) Chloromethyl 4-( 2-N-benzyloxycarbonyl-L-valyloxyethoxy)
1o benzoate.
To a solution of 4-(2- N-benzyloxycarbonyl-L-valyloxyethoxy) benzoic acid
(6.7g,
16.1 mmole) in 1,4-dioxane (80 ml} was added a 40% solution of
tetrabutylammonium hydroxide (9.74g, 15 mmole) and the mixturewas stirred 2
hours at room temperature. The mixture was evaporated under reduced pressure
and
~5 coevaporated two times with 1,4-dioxane and two times with toluene. The
dried
product was dissolved in dichloromethane (30 ml) and iodochloromethane (42.5g,
241 mmole) was added. The solution was stirred overnight at room temperature
and
evaporated under reduced pressure. About 150 ml ethyl actate were
added and the organic phase washed twice with water, dried with sodium sulfate
and
2o evaporated under reduced pressure. The product was isolated by silica gel
column
chromatography with hexane/ethyl acetate. Yield: 1.2g = 17%
e) Iodomethyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethoxy) benzoate.
To a solution of chloromethyl 4-(2-N-benzyloxycarbonyl-L-valyloxyethoxy)
25 benzoate (l.lg, 2.37 mmole) in dry acetone (40 ml) was added sodium iodide
(l.Sg,
10.0 mmole) and the mixture was stirred overnight at room temperature. The
mixture
was evaporated under reduced pressure and extracted with ethyl actate water.
The
organic phase was washed with a 5% sodium thiosulfate solution, dried with
sodium
sulfate and evaporated under reduced pressure.Yield: 1.3g = 98%.
'H-NMR (CDC13) 0.88 (d, 3H) 0.95 (d, 3H) 2.18 (m, 1H) 4.22 (m, 2H) 4.32 (d, d,
1H) 4.50 (m, 2H) 5.10 {s, 2H) 5.22 (d, 1H) 6.14 (s, 2H) 6.90 (d, 2H) 7.35(m,
7H)
7.98 (d, 2H)
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Example A-I-28
Iodomethvl 4-f 3-(Cbz-L-valyloxy)-1-propel]Ibenzoate
O
O O
N-
O
a) t-Butyl4-[3-(Cbz-L-valyloxy)-1-propyl]benzoate.
t-Butyl 4-(3-hydroxy-1-propyl)benzoate (Taylor et al .l. Org. Chem. 1995, 60,
7947)
(3.3 g, 14 mmol), N,N'-dicychlohexyl carbodiimide (3.3 g, 16.8 mmol), CBZ-L-
valine (4.2 g, 16.8 mmol) and N,N dimethylaminopyridine (0.85 g, 7 mmol) were
to dissolved in a minimum amount of dichloromethane and left at ambient
temperature
over-night. The slurry was then filtrated and the solvent evaporated.
Purification by
chromatography (dichloromethane-ether, 20:1) gave pure title compound (7 g,
100
%).
Z 5 b) Chloromethyl 4-[3-(Cbz-L-valyloxy)-1-propyl]benzoate
t-Butyl 4-[3-(Cbz-L-valyloxy)-1-propyl]benzoate (6.5 g, 13.8 mmol) was
dissolved
in dichloromethane (50 ml). Triflouroacetic acid (10 ml) was added and the
mixture
was left over-night. The solvent was then removed and the crude residue co-
evaporated with toluene and 1,4-dioxan and dried in vacuum over-night. The
dried
2o product was dissolved in 1,4-dioxane (100 ml) and tetrabutyl
ammoniumhydroxide
(40 w/w % in water) was added dropwise until the pH = 7-8. The solvent was co-
evaported with toluene four times, and then dissolved in dichloromethane (100
ml)
followed by addition of 100 g of molecular sieve (4~) and the mixture was
stirred
for 30 minutes. Chloroiodomethane (15 ml) was added and the reaction mixture
was
25 left for five hours. Filtration and purification by chromatography (hexane-
ethyl
acetate, 3:1 ) gave pure title product (3.3 g, 52 %).
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c) Iodomethyl4-[3-(cbz-L-valyloxy)-1-propyl]benzoate
Chloromethyl 4-[3-(Cbz-L-valyloxy)-1-propyl]benzoate (3 g, 6.5 mmol) and
sodium
iodide (4.9 g, 32 mmol) were dissolved in acetonitril (SO ml) and heated to 65
C for 4
hours. The solvent was removed under reduced pressure and the residue was
taken up
in dichloromethane and filtrated. Removal of the solvent and purification by
chromatography (dichloromethane-ether, 20:1) gave pure title product (2.4 g,
67 %).
'H NMR (CDC13 45 °C): 7.95 (d, 2H), 7.36-7.24 (m, 7H), 6.15 (s, 2H),
5.2 (br s, 1H),
i o 5.12 (s, 2H), 4.330-4.20 (m, 1 H), 4.15 (t, 2H), 2.74 (t, 2H), 2.2-2.1 (m,
1 H), 1.99 (m,
t 2H), 0.98 (d, 3H), 0.91 (d, 3H)
Example A-I-29
Iodomethvl 4-f 3-(Cbz-L-isoleucyloxv)-1-propvl]Ibenzoate
0 o~i
i
0 0
O
O
a) t-Butyl4-[3-(Cbz-L-isoleucyloxy)-1-propyl]benzoate.
Prepared from t-butyl 4-(3-hydroxy-1-propyl)benzoate (3.5 g, 14.8 mmol) by the
same procedure as described for Example A-I-28 to give the title compound (7
g,
97%).
b) Chloromethyl4-[3-(Cbz-L-isoleucyloxy)-1-propyl]benzoate.
Prepared from the material of step a) (6.9 g, 14.4 mmol) by the procedure
described
in Example A-I-28 to give the title compound (3.8 g, 55%) as a colourless oil.
c) Iodomethyl4-[3-(Cbz-L-isoleucyloxy)-1-propyl]benzoate.
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Prepared from the product of step b (3.26 g, 6.8 mmol) by the same procedure
as for
Example A-I-28 to give pure title compound. (3.46 g, 90 %).
'H NMR (CDC13 45 °C): 7.98-7.93 {m, 2H), 7.35-7.23 (m, 7H), 6.15 (s,
2H), 5.25-
5.15 (br s, 1H), 5.13 (s, 2H), 4.32 (m, 1H), 4.12 (t, 2H), 2.74 (t, 2H), 1.94
(m, 2H),
1.86 (m, 1H), 1.5-1.4 (m, 1H), 1.25-1.15 (m, 1H), 0.95-0.86 (m, 6H).
Example A-I-30
Iodomethvl 2-( N-benzylox carbonylamino-2-meth~propionvlox~
1 o methyl 1-2-methyl propionate
a) 2-benzyloxycarbonylamino-2-methyl propionic acid.
To a solution of sodium carbonate (13.3g, 125 mmole) in water (100 ml) was
added
2-amino-2-methylpropionic acid (S.Og, 50 mmole) and the mixture was stirred
for 30
minutes. The solution was cooled to about 10°C and a 50% solution of
benzyl
chloroformiate (20.Sg, 60 mmole) was added dropwise. The mixture was stirred
at
roomtemperature overnight and acidified with 2M hydrochloric acid. The mixture
was extracted two times with ethyl acetate. The organic phase was washed with
water, dried with sodium sulfate and evaporated under reduced pressure. The
product
2o was isolated by silica gel column chromatography with hexane/ethyl acetate.
Yield:
7.7g = 64%
b) 4-methoxybenzyl2-(2-benzyloxycarbonylamino-2-
methylpropionyloxymethyl)-2-methyl propionate.
To a cooled solution of 4-methoxybenzyl 2-(hydroxymethyl)-2-methyl propionate
(3.6g, 15 mmol), 4-dimethylaminopyridine (0.37g, 3 mmole) and 2-
benzyloxycarbonylamino-2-methylpropionic acid (3.8g, 16 m-mole) in
dichloromethane (60 ml) was added dicyclohexyl-carbodiimide (3.7g, 18 mmole)
and
the mixture was stirred overnight at room temperature.The mixture was cooled
and
3o the urethane was filtered. The solution was evaporated and 150 ml ethyl
acetate were
added, The organic phase was washed twice with 5% acetic acid, 5% sodium
hydrogencarbonate and water. The organic phase was dried with sodium sulfate
and
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evaporated under reduced pressure. The product was isolated by silica gel
column
chromatography with toluene / acetone. Yield: 6,2g =91
c) 2-(benzyloxycarbonylamino-2-methylpropionyloxymethyl )-2-
methylpropionic acid.
To a solution of 4-methoxybenzyl 2-(2-benzyloxycarbonylamino-2-
methylpropionyloxymethyl)-2methyl propionate (6,1 g, 13,3 mmole)
dichloromethane (50 ml) was added trifluoro acetic acid (10 ml) and the
mixture was
stirred 3 hours at room temperature. The solution was evaporated under reduced
1o pressure and coevaporated two times with toluene. The product was isolated
by
silica gel column chromatography with dichloromethane/methanol. Yield: 4.1g =
91%
'H-NMR (CDCI,) 1.22 (s, 6H) 1.52 (s, 6H) 4.13 (m, 2H) 5.07 (s, 2H) 5.44 (s,
1H)
i5 7.33 (m, SH)
d) Chloromethyl 2-(2-benzyloxycarbonylamino-2-
methylpropionyloxymethyl)-2-methyl propionate.
To a solution of 2-(2-benzyloxycarbonylamino-2-methylpropionyloxymethyl)-2-
2o methyl propionic acid (4.Og, 11.8 mmole) in 80 ml 1,4-dioxane was added a
40%
solution of tetrabutylammonium hydroxide (7.Sg, 11.5 mmole) and the mixture
was
stirred 2 hours at room temperature. The mixture was evaporated under reduced
pressure and co-evaporated two times with 1,4-dioxane and two times with
toluene.
The dried product was dissolved in 20 ml dichloromethane and iodochloromethane
25 (31.7g. 180 mmole) was added. The solution was stirred overnight at room
temperature and evaporated under reduced pressure. About 100 ml ethyl actate
were
added and the organic phase washed twice with water, dried with sodium sulfate
and
evaporated under reduced pressure. The product was isolated by silica gel
column
chromatography with toluene/acetone. Yield: 2.Og = 45%.
e) Iodomethyl 2-( N-benzyloxycarbonylamino-2-
methylpropionyloxymethyl)-2-methyl propionate.
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To a solution of chloromethyl 2-(N-benzyloxycarbonylamino-2-
methylpropionyloxymethyl)-2-methyl propionate (1.9g, 4.92 mmole) in 50 ml dry
acetone was added sodium iodide (3.Og, 20 mmole) and the mixture was stirred
overnight at room temperature. The mixture was evaporated under reduced
pressure
and extracted with ethyl actate water. The organic phase was washed with a 5%
sodium thiosulfate solution, dried with sodium sulfate and evaporated under
reduced
pressure.
Yield: 2,1 g.
to 'H-NMR (CDC13) 1.20 (s, 6H) 1.53 (s, 6H) 4.13 (s, 2H) 5.07 (s, 2H) 5.30 {s,
1H)
5.90 (s, 2H) 7.35 (m, SH)
Example A-I-31
Iodomethvl2-methvl-2-(N-benzvlox carbonyl-L-alanyloxvmethyl)pro~ionate
1S
The title compound (alternative nomenclature 2,2-dimethyl-3-(CBz-alanyloxy)-
propioic acid iodomethyl ester) is prepared in a similar manner as described
in
Example A-I-10 using N-CBz protected L-alanine in place of N-CBz-D-valine.
20 'H-NMR. (CDCI3): 7.33 (m, 5H), 5.91 (m, 2H), 5.11 (s, 2H), 4.38 (m, 1H),
4.14
(m, 2H), 1.40 (d, 3H), 1.21 (d, 6H).
Example A-I-32
Iodomethyl 3-(N-CBz-L-valvloxy)-2-metl~l-propionate
O
N-CBz-L-Val-O O~I
a) 3-(N-CBz-L-valyloxy)-2-methyl-1-propano I .
To a mixture of DCC (16.5 g, 80 mmol), DMAP (0.977 g, 8 mmol) and N-CBz-L-
3o valine {20.1 g, 80 mmol) in CHzCIz {400 mL)-at 0 °C, was added
dropwise a solution
of 2-methyl-1,3-propanediol (72.1 g, 800 mrnol) in CHZCIz (100 mL). After lh
at 0
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°C, the temperature of the reaction mixture was allowed to assume room
temperature
and then the mixture was stirred for 16h at room temperature. The mixture was
filtered through a glass filter and the solvent was removed under reduced
pressure.
The crude product was column chromatographed (silica gel, 5% MeOH in CHZCIz},
to give 20.3 g 3-(N-CBz-L-valyloxy)-2-methyl-1-propanol.
b) 3-(N-CBz-L-valyloxy)-2-methyl-propionic acid.
To a solution of Cr03 (4.29 g, 42.9 mmol) in HOAc (50 mL) at 20 °C,
was added
dropwise a solution of 3-(N-CBz-L-valyloxy)-2-methyl-1-propanol (4.61 g, 14.3
1 o mmol) in aceton (25 mL). After stirnng for 24h, water (300 mL) was added
and the
reaction mixture was extracted with CHZCIz (4x200 mL) . The combined organic
layers was concentrated under reduced pressure. The residue was column
chromatographed (silica gel, 5-10% MeOH in CHzCIz), to give 3.10 g of 3-(N-CBz-
L-valyloxy)-2-methyl-propionic acid.
'H-NMR (CDC13): I I.54 (br s, IH), 7.33 (s, SH), 5.48 (d, 1H), S.11 (s, 2H),
4.40-
4.00 (m, 3H), 2.95-2.70 (m, 1H), 2.25-2.05 (m, IH), 1.30-1.12 (m, 3H), 0.95
(d, 3H),
0.87 (d, 3H).
2o c) Chloromethyl3-(N-CBz-L-valyloxy)-2-methyl-propionate.
To a solution of 3-(N-CBz-L-valyloxy)-2-methyl-propionic acid (3.10 g, 9.2
mmol)
in dioxane (100 mL), was added dropwise a 40% aqueous solution of
tetrabutylammonium hydroxide (5.97 g). After stirring for 5 min, the solution
was
evaporated to dryness through co-evaporation with dioxane and toluene. The
residue
was dissolved in dichloromethane (100 mL) and then chloroiodomethane (6.70 mL,
92 mmol) was added and the solution was stirred for 6h at room temperature.
The
solution was concentrated under reduced pressure and the residue was shaken
with
hexane / ethyl acetate (1:1 v/v, 200 mL). The yellow crystalline solid was
filtered off
and the filtrate was washed with aqueous solution of sodium thiosulfate (0.1
M) and
3o the filtered through anhydrous sodium sulfate and evaporated to dryness.
The residue
was column chromatographed (silica gel, 0.5-2% MeOH in CHZC12), to give 1.90 g
of
chloromethyl 3-(N-CBz-L-valyloxy)-2-methyl-propionate.
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d) Iodomethyl 3-(N-CBz-L-valyloxy)-2-methyl-propionate.
To a solution of chloromethyl 3-(N-CBz-L-valyloxy)-2-methyl-propionate (1.84
g,
4.77 mmol) in acetonitrile (50 mL), was added sodium iodide (2.86 g, 19.0
mmol).
The solution was stirred for 3.5 h at 60 °C. The resulting suspension
was filtered and
the filtrate was evaporated. The residue was dissolved in CHZC12 and washed
with
aqueous sodium thiosulfate (0.1 M). The organic phase was dried (Na2S04) and
concentrated under reduced pressure. The crude product was column
chromatographed (silica gel, 1% MeOH in CHZCIz), to give 1.78 g of iodomethyl
3-
(N-CBz-L-valyloxy)-2-methyl-propionate.
'H-NMR (CDCl3): 7.35 (s, SH), 5.90 (s, 2H), 5.25 (d, 1H), 5.10 (s, 2H), 4.36-
4.18
(m, 3H), 2.94-2.73 (m, 1H), 2.26-2.08 {m, 1H), 1.28-1.18 (m, 3H), 0.96 (d,
3H), 0.87
(d, 3H).
IS
Example A-I-33
Iodometh~(N-CBz-L-t-butyl~lvcvloxy,)-2 2-dimeth~rlnropionate
O O~~
CBzHN
O O
a) 4-Methoxybenzyl 3-(N-CBz-L-t-butylglycyloxy)-2,2-
2o dimethylpropionate.
To a mixture of DCC (4.95 g, 24 mmol), DMAP (293 mg, 2.4 mmol) and N-CBz-L-
t-butylglycine (6.37 g, 24 mmol) in CHzCl2 (100 mL) at 0 °C, was added
dropwise a
solution of 4-methoxybenzyl 2,2-dimethyl-3-hydroxypropionate (2.86 g, I2 mmol)
in CHzCl2 (50 mL). After lh at 0 °C, the temperature of the reaction
mixture was
25 allowed to assume room temperature and then the mixture was stirred for 48h
at
room temperature. The mixture was filtered through a glass filter and the
solvent was
removed under reduced pressure. The crude product was column chromatographed
(silica gel, 1-2% MeOH in CHZC12), to give 5.60 g 4-methoxybenzyl 3-(N-CBz-L-t-
butylglycyloxy)-2,2-dimethylpropionate.
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b) 3-(N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionic acid
To a solution of 4-methoxybenzyl 3-(N-CBz-L-t-butylglycyloxy)-2,2-
dimethylpropionate (5.60 g, 11.5 mmol) in CHzCl2 (90 mL) at room temperature,
was
added trifluoroacetic acid (10 mL). After lh at room temperature, the reaction
mixture was concentrated under reduced pressure. The crude product was column
chromatographed (silica gel, 3--~S% MeOH in CHZC12), to give 2.70 g of 3-(N-
CBz-
L-t-butylglycyloxy)-2,2-dimethylpropionic acid.
'H-NMR (CDCl3): 11.2 (br s, 1H), 7.34 (s, SH), 5.44 (d, 1H), 5.11 (s, 2H),
4.25-4.03
to (m, 3H), 1.32-1.22 (m, 6H), 0.97 (s, 9H).
c) Chloromethyl 3-(N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionate.
To a solution of 3-(N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionic acid (2.70
g,
7.38 mmol) in dioxane (100 mL), was added dropwise a 40% aqueous solution of
15 tetrabutylammonium hydroxide (4.79 g). After stirnng for S min, the
solution was
evaporated to dryness through co-evaporation with dioxane and toluene. The
residue
was dissolved in dichloromethane (100 mL) and then chloroiodomethane (5.37 mL,
73.8 mmol) was added and the solution was stirred for 6h at room temperature.
The
solution was concentrated under reduced pressure and the residue was shaken
with
2o hexane / ethyl acetate (1:1 v/v, 200 mL). The yellow crystalline solid was
filtered off
and the filtrate was washed with aqueous solution of sodium thiosulfate (0.1
M) and
the filtered through anhydrous sodium sulfate and evaporated to dryness. The
residue
was column chromatographed (silica gel, 0.5-1% MeOH in CH2C12), to give 2.44 g
of
chloromethyl 3-(N-CBz-L-t-butylglycyloxy}-2,2-dimethylpropionate.
d) Iodomethyl3-(N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionate.
To a solution of chloromethyl 3-(N-CBz-L-t-butylglycyloxy)-2,2-
dimethylpropionate
(2.44 g, 5.90 mmol) in acetonitrile (SO mL), was added sodium iodide (3.54 g,
23.6
mmol). The solution was stirred for 3.S h at 60 °C. The resulting
suspension was
3o filtered and the filtrate was evaporated. The residue was dissolved in
CHZCIz and
washed with aqueous sodium thiosulfate (0.1 M). The organic phase was dried
(Na2S04) and concentrated under reduced pressure. The crude product was column
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chromatographed (silica gel, 1 % MeOH in CHzCIz), to give 2.61 g of iodomethyl
3-
(N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionate.
'H-NMR (CDCI,): 7.32 (s, SH), 5.89-5.82 (m, 2H), 5.40 (d, 1H), 5.07 (s, 2H),
4.17
(d, 1H), 4.11 (s, 2H), 1.22-1.19 (m, 6H), 0.95 (s, 9H).
Example A-I-34
N benzyloxycarbonvl-traps-4-(N be loxycarbonvl-L-valyloxy~ L~roline
iodometh, 1 ester.
O ~CBz
O
O
to CBzNH O'/I
(a) N benzyloxycarbonyl-traps-4-hydroxy-L-proline p-methoxybenzyl
ester.
Cesium carbonate ( 1.23 g, 3.78 mmol) was added to a stirred solution of N
benzyloxycarbonyl-traps-4-hydroxy-L-proline (2.00 g, 7.54 mmol) in 20 mL dry
15 DMF. After 15 min, p-methoxybenzyl chloride ( 1.25 mL, 9.21 mmol) was added
and
the mixture was stirred for 17 h. The solvent was evaporated under vacuum, and
the
residue was partitioned between EtOAc (100 mL) and water (50 mL).The organic
phase was washed with water (2 x 25 mL), dried over anhydrous Na2S04, and
concentrated. Flash column chromatography (silica gel, 2/1 EtOAc - petroleum
ether)
2o gave 2.69 g (92%) of the p-methoxybenzyl ester.
(b) N benzyloxycarbonyl-traps-4-(N benzyloxycarbonyl-L-valyloxy)-L-
proline p-methoxybenzyl ester.
A mixture of N benzyloxycarbonyl-traps-4-hydroxy-L-proline p-methoxybenzyl
25 ester (2.54 g, 6.59 mmol), N benzyloxycarbonyl-L-valine (1.82 g, 7.24
mmol), 4-
dimethylaminopyridine (90 mg, 0.7 mmol), and dicyclohexylcarbodiimide (1.64 g,
7.95 mmol) in 26 mL dry CHZCIz was stirred overnight. The slurry was filtered,
and
the filtrate was concentrated under vacuum to give an oil that was subjected
to flash
column chromatography (silica gel, 3/1 and~then 2/1 petroleum ether - EtOAc)
to
3o yield 3.70 g ( 91 %) of the title compound.
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(c) N benzyloxycarbonyl-traps-4-(N benzyloxycarbonyl-L-valyloxy)-L-
proline chloromethyl ester.
The p-methoxybenzyl group was removed by stirnng a solution of the compound
(2.47 g, 4.0 mmol) from step (b) and 8 mL CF,COOH in 40 mL CHZCl2 for 30 min.
The reaction mixture was concentrated under vacuum, coevaporating several
times
with more CHzCl1 and toluene. A second sample (1.14 g, 1.84 mmol) was treated
similarly. The crude products obtained from both samples were combined and
subjected to flash column chromatography (silica gel, 15% methanol in CHzCh to
to give 2.58 g of cream-colored solids containing N benzyloxycarbonyl-traps-4-
(N
benzyloxycarbonyl-L-valyloxy)-1.-proline (TLC 15/85 MeOH/CHZCIz Rf = 0.25).
This material ( 1.14 g) was dissolved in 10 mL dioxane. Tetrabutylammonium
hydroxide ( 1.15 mL of a 40 wt % solution in H20, 1.76 mmol) was added, the
mixture was evaporated to dryness, and the residue was coevaporated several
times
15 with toluene and, lastly, CHzCl2. The resulting. Q salt was stirred with
chloroiodomethane (1.30 mL, 17.8 mmol) in dry CHZCIz (20 mL) for 20 h. The
solvent was removed under vacuum, 25 mL of 2/1 petroleum ether - EtOAc was
added, and the precipitates which formed were filtered. Concentration of the
filtrate,
followed by flash column chromatography (silica gel, 4/1 and then 2/1
petroleum
2o ether - EtOAc) gave the chloromethyl ester (580 mg) as white solids.
(d) N benzyloxycarbonyl-traps-4-(N benzyloxycarbonyl-L-valyloxy)-L-
proline iodomethyl ester.
The chloromethyl ester (533 mg, 0.974 mmol) from step (c) and NaI (594 mg,
3.96
25 mmol) were refluxed in dry MeCN (10 mL) at 75 °C for 4 h. After
removal of solvent
under vacuum, the mixture was partitioned between 20 mL EtOAc and 10 mL water.
The organic phase was washed with S% NazSz03 and then brine, dried over
anhydrous NazS04, and concentrated. Flash column chromatography (silica gel,
3/1
petroleum ether - EtOAc) gave the iodomethyl ester (526 mg, 84%) as white
solids.
'H NMR (2S0 MHz, CDC1,) 8 0.86 and 0.94 (2d, 3H each, J= 6.8 Hz), 2.13 (m,
2H),
2.31 (m, 1H), 3.65-3.80 (m, 2H), 4.24 (m, 1H), 4.43 (m, 1H), 5.03-5.19 (m,
4H), 5.30
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(br s, 1H), 5.54 (br d, 1H), 5.70 and 5.83-5.97 (s and ABq, 2H total), 7.30-
7.32 (m,
1 OH).
Prodru~~s of the invention
EXAMPLE A-1
4-amino-1-hydroxvbutvlidene-1 1-bisphosphonic acid tri(2 meth 1
valvloxvmethyl~propionvloxvmeth 11 ester.
to a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid, tri(2-methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl) propionyl-
oxymethyl) ester and 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic acid, di(2-methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl)
propionyloxymethyl) ester.
15 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid is
prepared
by the methodology in Kieczykowski et al, J Org Chem 1995, 60, 8310-8312, and
the
4-amino group CBz protected as shown in US 5 227 506. To a solution thereof
(195
mg, 0.51 mmole) in dry N,N-dimethylformamide (2 m1), was added
diisopropylethyl-
amine (0.27 ml, 1.53 mmole), followed by an injection of a solution of
iodomethyl 2-
2o methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl) propionate (626 mg, 1.27
mmole) in N,N-dimethylformamide (2 mI). After stirnng under argon for 2,5 h at
room temperature, the solution was concentrated on rotavapor and treated with
ethyl
acetate (10 ml). Crystals were f Itered off and the filtrate was extracted
with brine
containing a small amount of sodium thiosulfate. The organic phase was
filtered
25 through anhydrous sodium sulfate and evaporated. The title compounds were
isolated
by silica gel column chromatography (0-4, 7-8, 20-30% ethanol in
dichloromethane).
Triester (70 mg). Rf ( 10%MeOH/ CHC13) 0.45. 'H-NMR (CDC13): 7.30 (m, 20H),
5.85-5.43 (m, 9H), 5.08 (m, 8H), 4.36-3.95 (m, 9H), 3.10 (m, 2H), 2.15-1.75
(m,
30 7H), 1.19 (s, 18H), 0.86 (m, 18H).3'P-NMR (CDC13+1%CD30D) (H3P04
reference):
8 23.8 (d), 11.8(d);
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Diester (185 mg), R~(10%MeOH/ CHC13) 0.10 (at the center of oval spot from
baseline). 'H-NMR (CDC13+1 %CD30D): 7.31 (m, 15H), 5.79-5.63 (m, 4H), 5.08 (m,
6H), 4.35-4.10 (m, 6H), 3.10 (m, 2H), 2.18-1.70 (m, 6H), 1.19 (m, 12H), 0.87
(m,
12H). 3'P-NMR (CDC13+1%CD,OD)(H3P04 reference): 8 16.6 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, tri(2-methyl-2-
(L-valyloxymethyl) propionyloxymethyl) ester.
A solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid tri(2-methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl) propionyloxymethyl)
to Ester (203 mg, 0.136 mmol) in methanol / ethyl acetate / acetic acid (2:1:1
v/v/v) (8.7
ml) was hydrogenated over a Pd-black catalyst (93 mg) at 40 psi of hydrogen
for 16
h. The suspension was filtered through Celite on a fine pore sized glassinter
and
washed with methanol/ethyl acetate (2:1 ). The filtrate was evaporated to
dryness in
vacuo and the title compound as the tetra acetate was obtained as a white
solid after a
1 s few co-evaporations with dioxane and hexane.
3'P-NMR (CDC13+5%CD,OD)(H3P04 reference): 8 23.1 (m), 11:1 (m).
Example A-2
20 4Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid dil2 methyl 2 (L
val loxYmethyl~ronionvloxvmethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid, di(2-
methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl) propionyloxymethyl) ester (130
25 mg, 0.112 mmol) was hydrogenated over Pd-black (48 mg) by the method of
Example A-1 b), to give the title compound as the triacetate as a white solid
(90 mg).
3'P-NMR (CDC13+5%CD30D)(H3P04 reference}: 8 16.2 (br, s).
3o Example A-3
4-Amino-1-hydroxvbutylidene-1 1-bisphOSphonic acid di(2 methyl 2 ~~L va~loxyl
propionyloxymethyll ester
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a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid, di(2-methyl-2-(N-benzyloxycarbonyl-L-valyloxy) propionyloxymethyl)
ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (306 mg,
0.80 mmole) was esterified by the method described in Example A-1-a. After
silica
gel column chromatography (2-4, 6-12, 15-20% ethanol in dichloromethane), the
pure fractions containing the title compound were pooled together and
evaporated.
The residue was then dissolved in ethyl acetate and the solution extracted
twice with
1o aqueous saturated sodium bicarbonate and then twice with 5% aqueous EDTA-
disodium salt. (116 mg of title compound). Rf. (20%MeOH/CHC13) 0.20 (at the
center of oval spot from baseline).
'H-NMR (CDC13+1%CD,OD): 7.28 (m, 15H), 5.60 (m, 4H), 5.05 (m, 6H), 4.13 (m,
15 2H), 3.09 (m, 2H), 2.19-1.72 (m, 6H), 1.49 (m, 12H), 0.89 (m, 12H). "P-NMR
(CDCl3+1 %CD30D)(H3P04 reference): 8 15.3 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, di(2-methyl-2-
(L-valyloxy) propionyloxymethyl) ester.
20 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid, di(2-
methyl-2-(N-benzyloxycarbonyl-L-valyloxy) propionyloxymethyl) ester (116 mg,
0.107 mmol) was hydrogenated over Pd-black (46 mg) by the method of Example A-
1-b, to give the title compound as the triacetate as a white solid (71 mg).
25 3'P-NMR (CDC13+S%CD,OD)(H3P04 reference): 8 14.9 (s).
Example A-4
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosnhonic acid di (2-~L valyloxy~ 3 methyl
~S)~+~utyr5rloxymethXl) ester.
a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-l,l-bisphosphonic
acid, di (2-(N-benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-
butyryloxymethyl)
ester.
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4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (383 mg, 1
mmole) was esterified by the method described in Example A-3-a to yield 184 mg
of
title compound. Rf (20%MeOH/CHC13) 0.20 (at the center of oval spot from
baseline).
'H-NMR (CDCl3+ 1 %CD30D): 7.27 (m, 15H), 5.62 (m, 4H), 5.15-4.72 (m, 8H),
4.32 (m, 2H), 3.08 (m, 2H), 2.16-1.73 (m, 6H), 0.88 (rn, 24H). "P-NMR
(CDCI,+1%CD30D)(H3P04 reference): 8 15.5 (s).
io b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, di (2-(L-
valyloxy)-3-methyl-(S)-(+)-butyryloxymethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid, di (2-(N-
benzyloxycarbonyl-L-valyloxy)-3-methyl-(S)-(+)-butyryloxymethyl) ester (184
mg,
0.166 mmol) was hydrogenated over Pd-black (71 mg) by the method of Example A-
i5 1-b, to give the title compound as the triacetate as a white solid (95 mg).
3'P-NMR (CDC13+5%CD30D)(H3P04 reference): b 14.6 (s).
Example A-5
20 4-amino-1-hvdroxvbutvlidene-1 1-bisph~ osnhonic acid mono 2 methyl 2 (L
val~ymethyl) nropionyloxymethyl~ ester.
a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid, tribenzyl mono (2-methyl-2-(N-benzyloxycarbonyl-L-valyloxymethyl)
25 propionyloxymethyl) ester.
To a solution of 4-benzyloxycarbonylamino-I-hydroxybutylidene-1,1-
bisphosphonic
acid (1.54 g, 4 mmole) in dry N,N-dimethylformamide (24 ml), heated at 50
°C, was
added diisopropylethylamine (2.78 ml, 16 mmole), followed by dropwise addition
of
benzylbromide ( 1.9 ml, 16 mmole). After stirring under argon for 4 h, the
solution
3o was concentrated on rotavapor and treated with ethyl acetate (20 ml).
Crystals were
filtered off and the filtrate was extracted with brine. The organic phase was
filtered
through anhydrous sodium sulfate and evaporated. The 4-benzyloxycarbonylamino-
1-hydroxybutylidene-1,1-bisphosphonic acid, tribenzylester was isolated by
silica geI
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column chromatography (2-4, 7-10, 15-20% ethanol in dichloromethane). The pure
fractions containing the pure triester were pooled together and evaporated.
The
residue was then dissolved in ethyl acetate and the solution extracted three
times with
2M aqueous solution of citric acid. Triester (990 mg); Rf (20%MeOH/ CHC13)
0.15
(at the center of oval spot from baseline);
3'P-NMR (CDC13) (H3P04 reference): 8 20.4(d), 13.3 (d); 'H-NMR (CDCI3): 7.35-
7.10 (m, 20H), 5.20-4.91 (m, 8H), 4.60 (br, 1H), 3.00 (m, 2H), 2.12-1.75 (m,
4H).
1o b) Dried tribenzyl ester (395 mg) was dissolved in dry N,N-
dimethylformamide (3 ml), followed by addition of diisopropylethylamine (99
ml)
and a solution of iodornethyl 2-methyl-2-(N-benzyloxycarbonyl-L-
valyloxymethyl)
propionate (737 mg) in N,N-dimethylformamide (1 ml). After stirring under
argon
for 4 h at 30 °C, the solution was concentrated to dryness on rotavapor
and treated
with ethyl acetate (10 ml). Crystals were filtered off and the filtrate was
extracted
with brine brine containing a small amount of sodium thiosulfate. The organic
phase
was filtered through anhydrous sodium sulfate and evaporated. The title
compound
(84 mg) was isolated by silica gel column chromatography (1, 2, 3% ethanol in
dichloromethane). Rf (2%MeOH/ CHCI3) 0.60;
3'P-NMR (CDC13) (H3P04 reference): 8 16.4(m). 'H-NMR (CDC13): 7.28 (m, 25H),
5.22 (d, 1H), 5.62-5.53 (m, 3H), 5.07, 5.04 (2xs, lOH), 4.93 (br, 1H), 4.27
(d,d, 1H),
4.1 S (d,d, 2H), 3.11 (m, 2H), 2.13-1.77 (m, SH), 1.17 (s, 6H), 0.92 (d, 3H),
0.83 (d,
3H).
c) 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid, mono (2-
methyl-2-(L -valyloxymethyl) propionyloxyrnethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid tribenzyl
mono (2-methyl-2-(N-benzyloxycarbonyl-L-valyloxyrnethyl) propionyloxymethyl)
3o ester (84 mg, 0.083 mmol) was hydrogenated over Pd-black (60 mg) by the
method
of Example A-1-b, to give the title compound as a white solid (35 mg).
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3'P-NMR (CDC13+5%CD30D)(H3P04 reference): 8 14.5 (m).
Example A-6
4-amino-1-hvdroxvbutvlidene-1 1-bis~hosphonic acid di l2-l-L valyloxy) 2
phenyl
DL-acetyloxvmethyll ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (192 mg,
0.5 mmole) was esterified with the corresponding iodomethyl ester by the
method
described in Example A-3-a to yield 72 mg of the title compound as the tri-N-
CBz
protected form, ready for deprotection. Rf (20%MeOH/CHC13) 0.20 (at the center
of
oval spot from baseline).
'H-NMR (CDCl3+ 1 %CD30D): 7.44-7.10 (m, 25H), 5.94 (m, 2H), 5.59 (m, 2H),
5.18-4.85 (m, 6H), 4.36 (m, 2H), 3.00 (m, 2H), 2.12-1.63 (m, 6H), 0.95 (m,
12H).
3'P-NMR (CDC13+1%CD,OD)(H3P04 reference): 8 15.5 (s).
Example A-7
4-amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di 111 3 di
valvloxy)prop,Yl 2
ox carbonyloxy methvll ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (141 mg,
0.37 mmole) was esterified from the corresponding iodomethyl ester by the
method
described in Example A-1-a to yield 90 mg of title compound as the tri CBz
protected form, ready for deprotection. Rf (10%MeOH/CHC13) 0.20 (at the center
of
oval spot from baseline). (153 mg of mixture of the diester and triester).
'H-NMR (CDC13+ 1 %CD30D) of title compound: 7.29 (m, 25H), 5.65 (m, 4H), 5.14-
4.85 (m, 12H), 4.45-4.05 (m, 12H), 3.11 (rn, 2H), 2.14-1.76 (m, 8H), 0.87 (m,
24H).
3'P-NMR (CDCl3+1 %CD,OD)(H3PO4 reference): 8 16.7 (s).
Example A-8
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di l2-L-valyloxYl DL
propionyloxymethyl) ester.
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4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (158 mg,
0.41 mmole) was esterified from the corresponding iodomethyl ester by the
method
described in Example A-3-a to yield 110 mg of the title compound as the tri N-
Boc
protected from, ready for deprotection. Rf (20%MeOH/CHCl3) 0.15 (at the center
of
oval spot from baseline).
'H-NMR (CDCI,+1%CD30D): 7.29 (m, 15H), 5.65 (m, 4H), 5.15-4.95 (m, 8H), 4.33
(m, 2H), 3.09 (m, 2H), 2.22-1.74 (m, 6H), 1.52 (m, 6H), 0.92 (m, 12H). 3'P-NMR
(CDC13+1 %CD30D)(H,P04 reference): 8 16.8 (s).
Example A-9
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di l5 ~L valYloxv) 2 2
dimethvlvalervloxXlneth Il ester
a) 4-Benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di-(S-(N-CBz-L-valyloxy)-2,2-dimethylvaleryloxymethyl) ester
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
acid (575 mg, 1.50 mmol) in DMF (10 mL), was added diisopropylamine (0.78 mL,
4.5 mmol), followed by an injection of a solution of give iodomethyl 5-(N-CBz-
L-
valyloxy)-2,2-dimethylvalerate (1.95 g, 3.75 mmol) in DMF (5 mL). After
stirnng
under argon for 1.5 h at room temperature, the solution was concentrated on
rotavapor and treated with ethyl acetate (100 mI,). Crystals were filtered off
and the
filtrate was extracted with brine containing a small amount of sodium
thiosulfate.
The organic phase was filtered through anhydrous sodium sulfate and
evaporated.
After silica gel column (silica gel, 4-X20% MeOH in CHZCl2), the pure
fractions
containing the title compound were combined and evaporated. The residue was
then
dissolved in ethyl acetate and the solution extracted twice with aqueous
saturated
sodium bicarbonate and then twice with 5% aqueous EDTA-disodium salt. The
ethyl
3o acetate phase was evaporated, to give 171 mg of 4-benzyloxy carbonylamino-1
hydroxybutylidene-1,1-bisphosphonic acid di-(5-(N-CBz-L-valyloxy)-2,2
dimethylvaleryloxymethyl) ester.
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'H-NMR (CDC13): 7.30 (br s, 15H), 5.85-5.25 (m, 4H), 5.20-4.95 (m, 6H), 4.30-
3.95 (m, 6H), 3.18-3.00 (m, 2H), 2.20-1.75 (m, 6H), 1.7-1.4 (m, 8H), 1.3-1.0
(s,
12H), 1.0-0.8 (m, 12H).
s "P-NMR (CDC13)(H,PO4 reference): 16.0 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid di-(5-(L-
valyloxy)-2,2-dimethylvaleryloxymethyl) ester.
to A solution of 4-benzyloxy carbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
acid di-(5-(N-CBz-L-valyloxy)-2,2-dimethylvaleryloxymethyl) ester (171 mg,
0.147
mmol) in methanol / ethyl acetate / acetic acid (2:1:1 v/v/v) (20 mL) was
hydrogenated over a Pd-black catalyst (30 mg) at 40 psi of hydrogen for 6 h.
The
suspension was filtered through celite and the filtrate was evaporated to
dryness
15 under reduced pressure, to give 95 mg of 4-amino-1-hydroxybutylidene-1,1-
bisphosphonic acid di-(5-(L-valyloxy)-2,2-dimethylvaleryloxymethyl) ester was
obtained as a white solid.
'H-NMR (CDCI,): 5.75-5.30 (m, 4H), 5.20-4.95 (m, 6H), 4.20-3.80 (m, 6H), 3.00-
20 2.80 (m, 2H), 2.20-1.40 (m, 14H), 1.3-1.0 (m, 12H), 1.0-0.8 (m, 12H).
3'P-NMR (CDC13+CD3pD)(H,P04 reference): 8 17.3 (br s)
Example A-10
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di-((2 (L valylo~)
25 ethox carbonvloxv) methyl) ester
a) 4-benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di-((2-(N-CBz-L-valyloxy)-ethoxycarbonyloxy) methyl) ester
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
3o acid (550 mg, 1.44 mmol) in DMF (10 mL), was added diisopropylamine (0.75
mL,
4.32 mmol), followed by an injection of a solution of 2-(N-CBz-L-valyloxy)-
ethyl
iodomethyl carbonate (1.40 g, 3.60 mmol) in DMF (5 mL). After stirnng under
argon for 1.5 h at room temperature, the solution was concentrated on
rotavapor and
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treated with ethyl acetate (100 mL). Crystals were filtered off and the
filtrate was
extracted with brine containing a small amount of sodium thiosulfate. The
organic
phase was filtered through anhydrous sodium sulfate and evaporated. After
silica gel
column (silica gel, 4-X20% MeOH in CHZC12), the pure fractions containing the
title
compound were combined and evaporated. The residue was then dissolved in ethyl
acetate and the solution extracted twice with aqueous saturated sodium
bicarbonate
and then twice with 5% aqueous EDTA-disodium salt. The ethyl acetate phase was
evaporated, to give 160 mg of 4-benzyloxy carbonylamino-1-hydroxybutylidene-
1,1-
bisphosphonic acid di-((2-(N-CBz-L-valyloxy}-ethoxycarbonyloxy) methyl) ester.
'H-NMR (CDC13): 7.29 (br s, 15H), 5.90-5.30 (m, 4H), S.1 S-4.90 (m, 6H), 4.50-
4.00 (m, 10H), 3.18-3.00 (m, 2H), 2.20-1.50 (m, 6H), 1.05-0.80 (m, 12H).
"P-NMR (CDCl3)(H,P04 reference): 16.5 (s}.
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid di-((2-(L-
valyloxy)-ethoxycarbonyloxy) methyl) ester.
A solution of 4-benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di-((2-(N-CBz-L-valyloxy)-ethoxycarbonyloxy) methyl) ester (160 mg, 0.147
mmol) in methanol / ethyl acetate / acetic acid (2:1:1 v/v/v) (20 mL) was
2o hydrogenated over a Pd-black catalyst (30 mg) at 40 psi of hydrogen for 7
h. The
suspension was filtered through celite and the filtrate was evaporated to
dryness
under reduced pressure, to give 100 mg of 4-amino-1-hydroxybutylidene-1,1-
bisphosphonic acid di-((2-(L-valyloxy)-ethoxycarbonyloxy) methyl) ester was
obtained as a white solid.
'H-NMR (CDC13): 5.80-5.40 (m, 4H), 4.70-4.05 (m, l OH), 4.4-4.2 (m, 4H), 3.00-
2.80 (m, 2H), 2.20-1.50 (m, 6H), 1.05-0.80 (m, 12H).
3'P-NMR (CDC13+CD30D)(H3P04 reference): b 17.5 (br s).
3o Example A-11
4-Amino-1-hvdroxvbutvliden-1 1-bisphosphonic acid bis j2 2 dimethvl 3 (D
valvloxyl-~ropion~vmethyl~! ester
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a) 4-Benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic acid bis
[2,2-dimethyl-3-(N-CBZ-D-valyloxy)-propionyloxymethylJ ester
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic
acid (382 mg, 1 mmole) and diisopropylethyl (0.43 ml, 2.5 mmole) in DMF (3 ml)
at
-40° C was added 2,2-dimethyl-3-(N-CBz-D-valyloxy)-propionic acid
iodomethyl
ester (1.23 g, 2.5 mmole) in DMF (4 ml). The reaction was kept at 0° C
for 2.5 hr
and then at 4° C for 18 hr. The reaction mixture was evaporated in
vacuo and ethyl
acetate (20 ml) was added. The precipitate was filtered off and the organic
phase was
to washed with sodium bicarbonate aqueous solution and dried. The product was
isolated with silica gel column chromatography. 125 mg.
'H-NMR (CDCI3): 7.31 (m, 15 H) 5.71 (m, 4 H) 5.58 (d, 2 H) 5.12 (s, 4H) 5.05
(s, 2H) 4.30 (dd, 2H) 4.12 (m, 4 H) 3.18 (m, 2H) 2.05 (m, 6 H) 0.92 (dd, I2
H).
3'P-NMR (CDC13): 15.1
b) 4-Amino-1-hydroxybutyliden-1,1-bisphosphonic acid bis [2,2-dimethyl-3-
(D-valyloxy)-propionyloxymethyl] ester.
4-Benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic acid bis [2,2-
dimethyl-3-(N-CBZ-D-valyloxy)-propionyloxymethyl) ester (130 mg) was dissolved
in a mixed solvent of EtOAc/MeOH/AcOH (6 ml/3 ml/1.5 ml). To the solution was
added palladium black (60 mg). The reaction was kept under hydrogen atmosphere
(40 psi) until sampling showed the complete deprotection of the
benzyloxycarbonyl
groups. The reaction mixture was filtered, and then dried and coevaporated
with
2s toluene and methanol, giving the titled product. 102 mg.
3'P-NMR ( CDC13 + CD30D ) : 14.1
Example A-12
4-Amino-I-h~droxybutvliden-1 1-bisnhosnhonic acid bis j4 (N CBz L valvloxv)
butanoyloxvmethyl]! ester
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a) 4-Benzyloxycarbonylamino-1-hydroxybutyliden-I,1-bisphos~t~ni~~tcitt'tiiS
[4-(N-CBz-L-valyloxy)-butanoyloxymethyl] ester.
4-Benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic acid (573 mg,
1.5 mmole) was dissolved in dioxane (10 ml). To the solution was added
tetrabutylammonium hydroxide (40 %, 2.43 ml, 3.75 mmole). The solution was
evaporated and coevaporated with DMF several times. The residue was dissolved
in
DMF (5 ml). To the solution was added 4-(N-CBz-L-valyloxy) butyric acid
iodomethyl ester ( 1.79 g, 3.75 mmole) in DMF (S ml) portionwise in one hour.
The
reaction was kept at room temperature for 3 hr and then evaporated in vacuo.
Later,
1 o ethyl acetate (20 ml) was added. The precipitate was filtered off and the
organic
phase was washed with sodium bicarbonate aqueous solution and dried. The
product
was isolated with silica gel column chromatography. 135 mg.
'H-NMR (CDC13):7.25 (m, 15 H) 5.60 (m, 6 H ) 5.05 (m, 8H) 4.30-3.90 (m, 6 H)
3.10 (m, 2 H) 2.50-1.80 (m, 14 H) 0.85 (m, I2 H).
3'P-NMR (CDC13): 13.7.
b) 4-amino-1-hydroxybutyliden-I,I-bisphosphonic acid bis 4-(L-
valyloxy)-butanoyloxymethyl] ester.
4-Benzyloxycarbonylamino-I-hydroxybutyliden-I,1-bisphosphonic acid bis [4-(N-
CBz-L-valyloxy)-butanoyloxymethyl] ester (100 mg) was dissolved in a mixed
solvent of EtOAc/MeOH/AcOH (6 ml/3 ml/1.5 ml). To the solution was added
palladium black (80 mg). The reaction was kept under hydrogen atmosphere (40
psi)
until sampling showed the complete deprotection of the benzyloxycarbonyl
groups.
The reaction mixture was filtered, and then dried and coevaporated with
toluene and
methanol, giving the titled product. 70 mg.
3'P-NMR (CD,OD): 17.7
3o Example A-13
4-amino-1-hydroxybutvliden-1 1-bisphosphonic acid di (3 (L valyloxy)
benzovlox~methyl) ester
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a) 4-B enzyloxycarbonylamino- I -hydroxybutyliden-1,1-bisphonic
acid, di-(3-(N-benzyloxycarbonyl-L-valyloxy) benzoyloxymethyl) ester.
To a solution of4-benzyloxycarbonylamino-1-hydroxybutyliden-1,1-biphosphonic
acid (0.59g, I.5 mmole) and diisopropylethyl-amine (0.64g, S mmole) in N,N-
dimethylformamide (40 ml) was added dropwise a solution of iodomethyl-3-(N-
benzyloxycarbonyl-L-valyloxy)-benzoate (2.2g, 4.3 mmole) in N,N-
dimethylformamide (5 ml). The mixture was stirred 2 hours at room temperature
under argon. The mixture was evaporated under reduced pressure. Ethyl acetate
(50
1o ml) was added and the mixture was filtered after 2 hours. The organic phase
was
washed twice with 5% sodium hydrogencarbonate solution and dried with sodium
sulfate. The product was isolated by silica gel column chromatography.
Yield: 0.23g = 15%
'H-NMR (CDCI3 + 5% CD30D) 0.89 (m, 12H) 1.58-2.28 (m, 6H) 2.92 (m, 2H)
4.26 (m, 2H) 5.00 (m, 6H) 5.46-6.02 (m, 4H) 6.78- 7.86 (8m, 23H)
3'P-NMR (CDCl3 + 5% CD30D) 16.5 (s)
b) 4-amino-1-hydroxybutyliden-1,1-bisphosphonic acid, di-(3-(L-
2o valyloxy) benzoyloxymethyl) ester.
Deprotection of the CBz groups of 4-benzyloxycarbonylamino-1-hydroxybutyliden-
1,1-bisphonic acid, di-(3-(N-benzyloxycarbonyl-L-valyloxy) benzoyloxymethyl)
ester using mild conditions as specified in Greene, "Protecting Groups in
Organic
Synthesis, (John Wiley & Sons, New York, 1981 ) yields the title compound.
Example A-14
4-Amino-1-hvdroxvbutvliden-1 1-bisnhosnhonic acid di l3 fL valvloxvl
~ropionyloxymethyl] ester
3o a) 4-Benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic
acid, di-(3-( N-benzyloxycarbonyl-L-valyloxy)-propionyloxymethyl) ester.
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutyliden-I,I-bisphosphonic
acid (0.88g, 2,5 mmole) and diisopropylethyl-amine (0.78g, 6 mmole) in N,N-
dimethylformamide {40 ml) was added dropwise a solution of iodomethyI-3-(N-
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benzyloxycarbonyl-L-valyloxy)-propionate (2.3g, 4.95 mmole) in N,N-
dimethylformamide (5 ml). The mixture was stirred 2 hours at room temperature
under argon and evaporated under reduced pressure. Ethyl acetate (50 ml) was
added
and the mixture was filtered after 2 hours. The organic phase was washed twice
with
S% sodium hydrogencarbonate solution and dried with sodium sulfate. The
product
was isolated by silica gel column chromatography.Yield: 0.19g = 8%
'H-NMR {CDC13 + 5% CD30D) 0.89 (m, 12H) 1.62-2.16 (m, 6H) 2.60 (m, 4H) 3.08
(m, 2H) 4.12 (m, 2H) 4.30 (m, 4H) 5.02 (m, 6H) 5.42-5.64 (m, 4H) 7.24 (m, 15H)
to "P-NMR (CDC13 + 5% CD30D) 16,9 (s)
b) 4-Amino-I-hydroxybutyliden-1,1-bisphosphonic acid, di-(3-(L-
valyloxy)-propionyloxymethyl) ester.
Deprotection ofthe CBz groups of4-benzyloxycarbonylamino-1-hydroxybutyliden-
1,1-bisphosphonic acid, di-(3-(N-benzyloxycarbonyl-L-valyloxy)-
propionyloxyrnethyl) ester using mild conditions as specified in Greene,
"Protecting
Groups in Organic Synthesis, (John Wiley & Sons, New York, 1981 ) yields the
title
compound.
2o Example A-15
4-amino-1-hydroxvbutvlidene-1 1-bisnhosnhonic acid di (4 lL va~lox~
benzoyloxvmethvll ester
a) 4-B enzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
2s acid, di-(4-(N-benzyloxycarbonyl-L-valyloxy) benzoyloxymethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (14I mg,
0.37 mmole) was esterified from the con esponding iodomethyl ester by the
method
described in Example A-3-a) to yield 55 mg of title compound. Rf
(20%MeOH/CHCI,) 0.15 (at the center of oval spot from baseline).
'H-NMR (CDC13+ I %CD30D): 7.82 (m, 4H), 7.29 (m, I 5H), 6.97 (m, 4H), 5.85 (m,
4H), 5.11 (m, 6H), 4.46 (m, 2H), 3.10 (m, 2H), 2.30-1.77 (m, 6H), 1.52 (m,
6H), 0.99
(m, 12H).
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3'P-NMR (CDCI,+1%CD30D)(H3P04 reference): 8 15.6(s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, di-(4-(L-
valyloxy) benzoyloxymethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid, di-(4-(N-
benzyloxycarbonyl-L-valyloxy) benzoyloxymethyl) ester is CBz deprotected using
mild conditions as prescribed in Greene, "Protecting Groups in Organic
Synthesis,
(John Wiley & Sons, New York, 1981 ) to yield the title compound.
to Example A-16
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di !3 !3 4 di !L va~loxv)
phenyl) propionvlox~nethyl~ ester
a) 4-benzyloxycarbonylamino-1-hydroxybutylidene- l , l -bisphosphonic
15 acid, di-(3-(3,4-di(N-benzyloxycarbonyl-L-valyloxy) phenyl) propionyloxy-
methyl)
ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (143 mg,
0.37 mmole) was esterified from the corresponding iodoemthyl ester by the
method
described in Example A-3-a) to yield 169 mg of the title compound. Rr
20 (20%MeOH/CHCI3) 0.15 (at the center of oval spot from baseline).
'H-NMR (CDCI3+ 1%CD30D): 7.40-6.85 (m, 31H), 5.62 (m, 4H), 5.02 (m, lOH),
4.43 (m, 4H), 3.10 (m, 2H), 2.84 (m, 4H), 2.61 (m, 4H), 2.35-1.73 (m, 8H),
1.52 (m,
6H), 0.99 (m, 24H). "P-NMR (CDCl3+1 %CD30D)(H3P04 reference): 8 14.3 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, di-(3-(3,4-di-
(L-valyloxy) phenyl) propionyloxymethyl) ester.
4-Benzyloxycarbonylamino-1-hydroxybutyIidene-1,1-bisphosphonic acid, di-{3-
(3,4-
di(N-benzyloxycarbonyl-L-valyloxy) phenyl) propionyloxy- methyl) ester
3o is CBz deprotected using mild conditions as prescribed in Greene,
"Protecting
Groups in Organic Synthesis, (John Wiley & Sons, New York, 198I) to yield the
title
compound.
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Example A-17
I21
4-Amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid tril3 lL valvloxyl 2 2
dimethvlpropoxycarbonvloxymethyl,fester
a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid tri(3-(N benzyloxycarbonyl-L-valyloxy)-2,2-dimethylpropoxycarbonyl
oxymethyl) ester.
To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid (174 mg, 0.45 mmol) in dry DMF (1 mL) were added diisopropylethylamine
(240 p,L, 1.38 mmol), followed by 3-(N benzyloxycarbonyl-L-valyloxy)-2,2-
1o dimethylpropyl iodomethyl carbonate (592 mg, 1.14 mmol). After stirring for
5 h at
ambient temperature, under nitrogen, the reaction mixture was concentrated on
a
rotavapor, treated with ethyl acetate (15 mL), and filtered. The organic
solution was
washed with 5% NazS203, followed by brine. Drying over anhydrous NazS04 and
concentration gave a yellow oil that was subjected to column chromatography
(silica,
15 2/1 petroleum ether - ethyl acetate, 2.5 - 20% methanol in CHZCIz) to give
fine,
white solids (147 mg) enriched in the triester. The solids were dissolved in
ethyl
acetate, washed twice with 5% aqueous EDTA-disodium salt, dried over anhydrous
NazSO,, and evaporated to dryness under vacuum.
Rf (10% methanol in CHzCl2) 0.30; ' H NMR (250 MHz, CDCl3 + 1 % CD30D) 8
20 0.85-0.95 (m, 36 H), 1.70-2.20 (m, 7H), 3.10 (br s, 2H), 3.85-3.95 (br, 12
H), 4.25
(m, 3H), 5.05 (s, 8H), 5.52-6.0 (m, lOH), 7.30 (s, 20H); 3'P NMR (101 MHz,
CDC13
+ 1% CD30D) 8 13.6 and 24.6 (2d, .I= 47 Hz). The diester was a minor
component:
"p NMR 8 18.6 (s).
b) Removal of benzyloxycarbonyl protecting groups.
25 The triester (110 mg} from step (a) was hydrogenated at 40 psi over Pd
black (14 mg)
in 4.2 mL solvent (2/1/1 ethyl acetate - methanol - acetic acid) for 18.5 h.
The
suspension was filtered through a small column of celite and washed with ethyl
acetate - methanol. The filtrate was evaporated to dryness under vacuum to
give
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white solids (97 mg). Because proton NMR showed incomplete deprotection, the
material was resubmitted for hydrogenation (14 mg Pd, 10 mL solvent) overnight
to
give the deprotected triester (as the acetate salt, 79 mg) as shown by the
disappearance of peaks for the benzyloxy group at 8 5.05 and 7.30.
"P NMR ( 1 O 1 MHz, CDC13) 8 12.5 and 24Ø
Example A-18
4-Amino-1-hvdroxvbutvlidene-1 1-bisnhosnhonic acid di j2-methyl 1 ~ valvloxvl
2-propoxycarbonYloxymethyll ester
a) 4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid
to di(1-(N benzyloxycarbonyl-L-valyloxy)-2-methyl-2-propoxycarbonyloxymethyl)
ester.
4-Benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (460 mg,
1.2 mmol) was esterified by the method described in Example A-17 a) with 1-(N
benzyloxycarbonyl-L-valyloxy)-2-methyl-2-propyl iodomethyl carbonate (1.54 g,
3.0
15 mmol) for 2h. After column chromatography (silica, 4-20% methanol in
CHZC12), the
pure fractions containing the title compound were pooled together and
concentrated.
The residue was dissolved in ethyl acetate, washed twice with 5% aqueous EDTA-
disodium salt, and then, water, dried over NazS04, and evaporated to dryness
under
vacuum to give the diester as off white solids (92 mg).
20 "P NMR (101 MHz, CDC13 + 1% CD30D) 8 19.5 (s);'H NMR (250 MHz, CDCl3 +
1% CD30D) 8 0.76-1.41 (m, 24H), 1.74 (br s, 4H), 2.05 (m, 2H), 3.02 (br s,
CHIN),
3.90-4.30 (m, CHZOC=O and CHa valine), 4.93-5.01 (m, 6H), 5.30-5.90 (m, OCH~O
and NHC=O), 7.22 (s, 15H).
(b ) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid, di(2-methyl-1-
25 (L-valyloxy)-2-propoxycarbonyloxymethyl) ester.
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The benzyloxycarbonyl-protected diester (86 mg) from step (a) was hydrogenated
by
the method described in Example A-17 b) to give the title compound (as the
acetate
salt) as a white powder (72 mg).
3'P NMR (101 MHz, CDC13) b 19.2 (s).
Example A-19
4-amino-1-hydroxybutylidene-1 1-bisphosphonic acid di (2 methyl 2 (L
isoleuc~ox~!nneth~l)propion~loxvmethvll ester.
a) 4-N-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di (2-methyl-2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl)propionyl-
oxymethyl) ester.
To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid (824mg, 2.1 mmole) and diisopropylethylamine (0.8g, 6.3 mmole) in dry N,N-
dimethylformamide (15 ml) was added dropwise a solution of iodomethyl 2-methyl-
is 2-(N-benzyloxycarbonyl-L-isoleucyloxymethyl) propionate (3.1g, 5.21 mmole)
in
N,N-dimethylformamide (6 ml). The mixture was stirred 2 hours at room
temperature
and evaporated under reduced pressure. Ethyl acetate (70 ml) was added and
after 1
hour the crystals were filtered. The organic phase was washed two times with
saturated sodium hydrogencarbonate solution, dried with sodium sulfate and
2o evaporated under reduced pressure. The product was isolated by silica gel
column
chromatography with dichloromethane/methanol.Yield: 0.36g
'H-NMR (CDCl3) 0.86(m, 12H) 1.20 (m, 16H) 1.60-2.20 (m, 6H) 3.10 (m, 2H) 3.80-
4.40 (m, 6H) 5.08 (m, 6H) 5.45 (m, 4H) 7.29 (m, 15H)
25 "P-NMR (CDCl3 +S~~o CD30D) 14.2 (s)
b) 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid di (2-methyl-
2-(isoleucyloxymethyl)propionyloxymethyl) ester.
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
3o acid di (2-methyl-2-(N-benzyloxycarbonylamino-L-
isoleucyloxymethyl)propionyl-
oxymethyl) ester (0.195g, 0.171 mmole) in ethyl acetate (10 ml), methanol (10
ml)
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and acetic acid (S ml) was added palladium black (100mg). The mixture was
hydrogenated overnight at 45 psi. The catalyst was filtered and washed with
ethyl
acetate, methanol and acetic acid. The solution was evaporated under reduced
pressure and the product was dried in vacuo to yield the title compound as the
triacetate salt. Yield: 150mg.
"P-NMR (CDC13 +S% CD30D) 18.1 (s)
Example A-20
4-amino-1-hydroxybutvlidene-1 1-bisphosnhonic acid di (4 fL valyloxyl
~yclohexano Ioxmethyl) ester
a) 4-N-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di (4-(N-benzyloxycarbonyl-L-valyloxy)-cyclohexanoyloxymethyl) ester.
To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-biphosphonic
acid (0.706g, 1.8 mmole) and diisopropylethylamine (07g, 5.4 mmole) in N,N-
dimethylformamide (15 m1) was added dropwise a solution of iodomethyl 4-(N-
benzyloxycarbonyl-L-valyloxy)-cyclohexanoate (2.35g, 4.5 mmole) in N,N-
dimethylformamide (5 ml). The mixture was stirred 2 hours at room temperature
2o under argon. The mixture was evaporated under reduced pressure. Ethyl
acetate (60
ml) was added and the solid was filtered after 2 hours. The organic phase was
washed
twice with saturated sodium hydrogencarbonate and brine. The organic phase was
dried with sodium sulfate and evaporated under reduced pressure. The
products were isolated by silica gel column chromatography with
dichloromethane/
methanol. After silica gel column chromatography the fractions were dissolved
in
ethyl acetate and washed three times with 5% aqueous EDTA-disodium salt
solution,
dried with sodium sulfate, evaporated under reduced pressure and dried in
vacuo to
yield 298mg
'H-NMR (CDC13) 0.84 (m, 12H) 1.35-2.35 (m, 24H) 3.10 (m, 2H) 4.08 (m, 2H) 5.02
(m, 8H) 5.55 (m, 4H) 7.24 (m, 15H)
3'P-NMR (CDCI3 + S% CD30D) 15.2 (s) -
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b) 4-amino-I-hydroxybutylidene-1,1-bisphosphonic acid di (4-(L-
valyloxy)-cyclohexanoyloxymethyl) ester.
4-N-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di (4-(N-benzyloxycarbonyl-L-valyloxy)-cyclohexanoyloxymethyl) ester is
de-
CBz protected using conventional conditions as exemplified above to yield the
title
compound.
Example A-21
4-amino-I-hydroxvbutvlidene-1 1-bisphosnhonic acid tri l4 (L valvloxy,~
to cvclohexano loxvmethvll ester
The appropriate fraction of Example A-20, step a) was dissolved in ethyl
acetate and
washed three times with 5% aqueous EDTA-disodium salt solution, dried with
sodium sulfate, evaporated under reduced pressure and dried in vacuo to yield
320
15 mg which is deprotected using conventional conditions as exemplified above
to yiled
the title compound.
'H-NMR (protected form) (CDCl3) 0.86 (m, 18H) I.3-2.5 (m, 34H) 3.10 (m, 2H)
4.22 (m, 3H) 4.55-5.10 (m, 11H) 5.50 (m, 6H) 7.28 (m, 20H)
20 "P-NMR (CDCI, + 5% CD30D) 20.8 (d) 10.0 (d)
Example A-22
4amino-1-hvdroxvbutvlidene-1 1-bisph_ osnhonic acid di 11V (2 L val ~~loxv 1 1
dimethvl-ethyl)aminocarbonvloxvmethvllester.
a) 4-N-benzyloxycarbonylamino- I -hydroxybutylidene-1,1-bisphosphonic
acid di [N-(2-L-valyloxy-1,1-dimethyl-ethyl)aminocarbonyloxymethyl]ester.
To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidenel,l-biphosphonic
aid (O.lg, 0. mmole) and diisopropylethylamine (0.31g, 2.4 mmole) in N,N-
3o dimethylformamide (5 ml) was added dropwise a solution of 2-(N-(
iodomethoxy-
carbonyl)-amino)-2-methyl-1-(N-benzyloxycarbonyl-L-valyloxy)-propane in N,N-
dimethylformamide (2.5 ml). The mixture was stirred 2 hours at room
temperature
under argon. The mixture was evaporated under reduced pressure. Ethyl acetate
(40
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ml) was added and the solid was filtered after 2 hours. The organic phase was
washed
twice with saturated sodium hydrogencarbonate and brine. The organic phase was
dried with sodium sulfate and evaporated under reduced pressure. The product
was
isolated by silica gel column chromatography with dichloromethane/methanol
acetic
acid. After silica gel column chromatography the fractions were dissolved in
ethyl
acetate and washed three times with 5% aqueous EDTA-disodium salt solution,
dried
with sodium sulfate, evaporated under reduced pressure and dried in vacuo.
Yield: 165mg.
'H-NMR (CDC13) 0.80 (m, 12H) 1.20 (m, 12H) 1.88 (m, 6H) 3.00 (m, 2H) 3.64 (m,
4H) 4.20 (m, 2H) 5.00 (m, 6H) 5.50 (m, 4H) 7.26 (m, 15H)
"P-NMR (CDCl3 + 5% CD30D) 14.8 (s)
b) 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid di [N-(2-L-
valyloxy-1,1-dimethyl-ethyl)aminocarbonyloxymethyl] ester.
A solution of 4-N-benzyloxycarbonylamino-1-hydroxybutyliden-1,1-bisphosphonic
acid di[N-(2-L-valyloxy-1,1-dimethyl-ethyl)aminocarbonyloxymethyl] ester
(0.16g
0.14mmole) in ethyl acetate (Sml), methanol (10 ml) and acetic acid (5 ml) was
hydrogenated with palladium black O.lg) at room temperature and 45 psi
overnight .
2o The catalyst was filtered and washed with ethyl acetate and methanol. The
solution
was evaporated under reduced pressure and the product was dried in vacuo to
yield
the triacetate salt. Yield: 100mg
3'P-NMR (CDCl3 + 5% CD30D) 14.7 (s)
Example A-23
4-amino-1-h~droxvbutylidene-1 1-bisphOSDhonic acid di 1 (2 L valylox~th ly-) 6
oxo-1.6-dihvd. ro-nyridine-3-carbonyloxymeth~) ester
3o a) 4-Benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di-(1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
carbonyloxymethyl) ester.
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To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid (565 mg, 1.44 mmol) in DMF (10 mL), was added diisopropylamine (0.75 mL,
4.32 mmol), followed by an injection of a solution of give 1-(2-N-CBz-L-
valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carboxylic acid iodomethyl ester
(2.00
g, 3.60 mmol) in DMF (5 mL). After stirring under argon for 1.5 h at room
temperature, the solution was concentrated and treated with ethyl acetate (100
mL).
Crystals were filtered off and the filtrate was extracted with brine
containing a small
amount of sodium thiosulfate. The organic phase was filtered through anhydrous
sodium sulfate and evaporated. After silica gel column (silica gel, 4-a20%
MeOH in
1o CHZCIZ), the pure fractions containing the title compound were combined and
evaporated. The residue was then dissolved in ethyl acetate and the solution
extracted
twice with aqueous saturated sodium bicarbonate and then twice with 5% aqueous
EDTA-disodium salt. The ethyl acetate phase was evaporated, to give 205 mg of
4-
benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid di-(1-(2-N-
CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester.
'H-NMR (CDCI3): 8.35-7.95 (m, 2H), 7.85-7.50 (m,2H), 7.26 (br s, 15H), 6.60-
6.20
(m,2H), 5.90-5.35 (m, 4H), 5. i 5-4.80 (m, 6H), 4.50-4.00 (m, l OH), 3.18-3.00
(m,
2H), 2.45-1.55 (m, 6H), 1.00-0.80 (m, 12H).
3'P-NMR (CDCI,)(H3P0, reference): 16.8 (s).
b) 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid di-(1-(2-L-
valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-carbonyloxymethyl) ester.
A solution of 4-benzyloxy carbonylamino-1-hydroxybutylidene-l,l-bisphosphonic
2s acid di-(1-(2-N-CBz-L-valyloxyethyl)-6-oxo-1,6-dihydro-pyridine-3-
carbonyloxymethyl) ester ( 180 mg, 0. i 45 mmol) in methanol / ethyl acetate /
acetic
acid (2:1:1 v/v/v) (20 mL) was hydrogenated over a Pd-black catalyst (30 mg)
at 40
psi of hydrogen for 10 h. The suspension was filtered through celite and the
filtrate
was evaporated to dryness under reduced pressure, to give 85 mg of 4-amino-1-
3o hydroxybutylidene-1,1-bisphosphonic acid di-(1-(2-L-valyloxyethyl)-6-oxo-
1,6
dihydro-pyridine-3-carbonyloxymethyl) ester was obtained as a white solid.
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'H-NMR (CDC13): 8.65-8.20 (m, 2H), 7.95-7.65 (m, 2H), 6.65-6.30 (m, 2H), 5.90-
5.35 (m, 4H), 4.70-4.00 (m, lOH), 3.18-3.00 (m, 2H), 2.45-1.55 (m, 6H), 1.00-
0.80
(m, 12H).
3'P-NMR (CDC13)(H3P04 reference): 13.9 (s).
Example A 24
4-Amino-1-hydroxvbutvlidene-1 1-bisnhosphonic acid di (fl 3 di
(valvloxvlnronvl
2-oxycarbonvloxy methyl) ester
to a) 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di ((1,3-di-(N-benzyloxycarbonyl)valyloxy)propyl-2-oxycarbonyloxy methyl)
ester.
4-benzyloxycarbonylamino-1-hydroxybutylidene-l,l-bisphosphonic acid (141 mg,
0.37 mmole) was esterified by the method described in Example A-1 a using the
1 s corresponding iodomethyl ester to give 90 mg of the title compound.
Rf (10%MeOH/CHC13) 0.20 (at the center of oval spot from baseline).
'H-NMR (CDC13+ 1 %CD30D): 7.29 (m, 25H), 5.65 (m, 4H), 5.14-4.85 (m, 12H),
4.45-4.05 (m, 12H), 3.11 (m, 2H), 2.14-1.76 (m, 8H), 0.87 (m, 24H). "P-NMR
20 (CDC13+1 %CD30D)(H,P04 reference): 8 16.7 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid di ((1,3-di-
(valyloxy)propyl-2-oxycarbonyloxy methyl) ester.
A solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
25 acid di ((1,3-di-(N-benzyloxycarbonyl)valyloxy)propyl-2-oxycarbonyloxy
methyl)
ester (162 mg, 0.1 mmol) in methanol / ethyl acetate / acetic acid (2:1:1
v/v/v) (6 ml)
was hydrogenated over a Pd-black catalyst (70 mg) at 40 psi of hydrogen for 16
h.
The suspension was filtered through Celite on a fine pore sized glass sinter
and
washed with methanol/ethyl acetate (2:1). The filtrate was evaporated to
dryness in
3o vacuo and the title compound was obtained as the pentacetate, a white solid
after a
few co-evaporations with dioxane and hexane.
3'P-NMR (CDCl3+ 5%CD30D)(H3P04 reference): b 13.5 (br).
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Example A 25
129
4-Amino-1-hvdroxvbutyliden-1 1-bisnhosphonic acid di~2 lL valvloxymethyl) 2
ethyl butvrovloxymethyl) ester
a) 4-N-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di-(2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-ethyl )-butyroyloxymethyl)
ester.
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutyliden-1,1-biphosphonic
acid (0.777g, 2.0 mmole) and diisopropylethylamine (0.78g, 6.0 mmole) in N,N-
to dimethylformamide (15 ml) was added dropwise a solution of iodomethyl 2-(N-
benzyloxycarbonyl-L-valyloxymethyl)-2-ethyl butyrate (2.6g, 5 mmole) in N,N-
dimethylformamide (5 ml). The mixture was stirred 2 hours at room temperature
under argon. The mixture was evaporated under reduced pressure. Ethyl acetate
(60
ml) was added and the solid was filtered after 2 hours. The organic phase was
washed
1 s twice with saturated sodium hydrogencarbonate and brine. The organic phase
was
dried with sodium sulfate and evaporated under reduced pressure. The products
were
isolated by silica gel column chromatography with dichloromethane/methanol.
After
silica gel column chromatography the~pure fractions were dissolved in ethyl
acetate
and washed three times with S% aqueous EDTA-disodium salt solution, dried with
2o sodium sulfate, evaporated under reduced pressure and dried in vacuo.Yield:
320mg
'H-NMR (CDC13) 0.84 (m, 24H) 1.5-2.2 (m, 14H) 3.10 (m, 2H) 4.20 (m, 2H) 5.0
(m, 6H) 5.50 (m, 4H) 7.26 (m, 15H) 3'P-NMR (CDC13 + 5% CD30D) 14.5 (s)
2s b) 4-amino-1-hydroxybutyliden-1,1-bisphosphonic acid di-(2-(L-
valyloxymethyl)-2-ethyl butyroyloxymethyl) ester.
A solution of4-benzyloxycarbonylamino-1-butylidene-1,1-bisphosphonic acid di-
(2-(N-benzyloxycarbonyl-L-valyloxymethyl)-2-ethyl butyroyloxymethyl) ester
(0.195g, 0.167 mmole) in methanol (15 ml), ethyl acetate (5 ml) and acetic
acid (10
3o ml) was hydrogenated with palladium black (120mg) at room temperature
overnight
with 45 psi. The catalyst was filtered and washed with ethyl acetate, methanol
and
acetic acid. The solution was evaporated under reduced pressure and solidified
with
ether. The product was filtered and dried in vacuo to yield the tri acetate
salt.
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130
Yield: 0.125g. "P-NMR (CDC13 + 5% CD30D) 14.9 (s)
Example A 26
4-Amino-1-hvdroxybutvlidene-1 1-bisphosphonic acid bis l3 fL valvloxvl 2
methvl-propionvloxymethvl) ester
a) 4-Benzyloxy carbonylamino-I-hydroxybutylidene-I,1-bisphosphonic
acid bis (3-(N-CBz-L-valyloxy)-2-methyl-propionyloxymethyl) ester.
to To a solution of4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
acid (580 mg, 1.48 mmol) in DMF (10 mL), was added diisopropylamine (0.773 mL,
4.44 mmol), followed by an injection of a solution of iodomethyl 3-(N-CBz-L-
valyloxy)-2-methyl-propionate ( 1.77 g, 3.70 mmol) in DMF (5 mL). After
stirring
under argon for 1.5 h at room temperature, the solution was concentrated on
rotavapor and treated with ethyl acetate (100 mL). Crystals were filtered off
and the
filtrate was extracted with brine containing a small amount of sodium
thiosulfate.
The organic phase was filtered through anhydrous sodium sulfate and
evaporated.
After silica gel column (silica gel, 4-X20% MeOH in CHZCIz), the pure
fractions
containing the title compound were combined and evaporated. The residue was
then
2o dissolved in ethyl acetate and the solution extracted twice with aqueous
saturated
sodium bicarbonate and then twice with 5% aqueous EDTA-disodium salt. The
ethyl
acetate phase was evaporated, to give 189 mg of 4-benzyloxy carbonylamino-1-
hydroxybutylidene-1,1-bisphosphonic acid bis (3-(N-CBz-L-valyloxy)-2-methyl-
propionyloxymethyl) ester.
'H-NMR (CDC13+1 %CD30D): 7.32 (br s, I SH), 5.85-5.30 (m, 4H), 5.18-4.93 (m,
6H), 4.30-3.95 (m, 6H), 3.22-3.00 (m, 2H), 2.93-2.70 (m, 2H), 2.20-1.65 (m,
6H),
1.35-1.05 (m, 6H), 1.00-0.75 (m, 12H).
3'P-NMR (CDCl3)(H3P04 reference): 15.1 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid bis (3-(L-
valyloxy)-2-methyl-propionyloxymethyl) ester.
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A solution of4-benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid bis (3-(N-CBz-L-valyloxy)-2-methyl-propionyloxymethyl) ester (189 mg,
0.175
mmol) in methanol / ethyl acetate / acetic acid (2:1:1 v/v/v) (20 mL) was
hydrogenated over a Pd-black catalyst (35 mg) at 50 psi of hydrogen for 7h.
The
suspension was filtered through celite and the filtrate was evaporated to
dryness
under reduced pressure, to give 110 mg of 4-amino-1-hydroxybutylidene-1,1-
bisphosphonic acid bis (3-(L-valyloxy)-2-methyl-propionyloxymethyl) ester as
the
acetate salt, a white solid.
3'P-NMR (CDCI,+CD30D)(H3P04 reference): & 14.7 (br s)
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Example A 27
132
4-Amino-1-hvdroxvbutvlidene-1 1-bisnhosphonic acid bis l3 (L t butvlQlvcvloxv)
2 2-dimethvlnronionvloxvmethvl) ester
a) 4-Benzyloxy carbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid bis (3-( N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionyloxymethyl) ester.
To a solution of 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-
bisphosphonic
acid (588 mg, 1.50 mmol) in DMF (10 mL), was added diisopropylamine (0.783 mL,
4.50 mmol), followed by an injection of a solution of iodomethyl 3-(N-CBz-L-t-
1o butylglycyloxy)-2,2-dimethylpropionate (1.90 g, 3.75 mmol) in DMF (5 mL).
After
stirring under argon for 1.5 h at room temperature, the solution was
concentrated on
rotavapor and treated with ethyl acetate (100 mL). Crystals were filtered off
and the
filtrate was extracted with brine containing a small amount of sodium
thiosulfate.
The organic phase was filtered through anhydrous sodium sulfate and
evaporated.
After silica gel column (silica gel, 520% MeOH in CHZCIz), the pure fractions
containing the title compound were combined and evaporated. The residue was
then
dissolved in ethyl acetate and the solution extracted twice with aqueous
saturated
sodium bicarbonate and then twice with 5% aqueous EDTA-disodium salt. The
ethyl
acetate phase was evaporated, to give 158 mg of 4-benzyloxy carbonylamino-1-
2o hydroxybutylidene-1,1-bisphosphonic acid bis (3-( N-CBz-L-t-butylglycyloxy)-
2,2-
dimethylpropionyloxymethyl) ester.
'H-NMR (CDCl3+1 %CD30D): 7.26 (br s, 1 SH), 5.85-5.40 (rn, 4H), S.15-4.88 (m,
6H), 4.20-3.95 (m, 6H), 3.20-3.00 (m, 2H), 2.10-1.65 (m, 4H), 1.22-1.05 (m,
12H),
1.00-0.75 (m, 18H).
3'P-NMR (CDCI3)(H3P04 reference): 15.1 (s).
b) 4-Amino-1-hydroxybutylidene-1,1-bisphosphonic acid bis (3-( L-t-
butylglycyloxy)-2,2-dimethylpropionyloxymethyl) ester.
3o A solution of 4-benzyloxy carbonylamino-1-hydroxybutylidene-l,l-
bisphosphonic
acid bis (3-( N-CBz-L-t-butylglycyloxy)-2,2-dimethylpropionyIoxymethyl) ester
(158 mg, 0.139 mmol) in methanol / ethyl acetate / acetic acid (2:1:1 v/v/v)
(20 mL)
was hydrogenated over a Pd-black catalyst {40 mg) at 50 psi of hydrogen for
16h.
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The suspension was filtered through celite and the filtrate was evaporated to
dryness
under reduced pressure, to give 100 mg of 4-amino-1-hydroxybutylidene-1,1-
bisphosphonic acid bis (3-( L-t-butylglycyloxy)-2,2-
dimethylpropionyloxymethyl)
ester as a white solid.
3'P-NMR (CDCl3+CD3pD)(H3P04 reference): 8 14.6 (br s)
Example A 28
4-amino-1-hvdroxvbutvlidene-1 1-bisphosphonic acid di~trans 4 (L valvloxvl L
1 o prolyoxvmeth~] ester
a) 4-benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic
acid di[N benzyloxycarbonyl-traps-4-(N benzyloxycarbonyl-L-valyloxy)-L-
prolyoxymethyl] ester.
15 Diisopropylethylamine (195 p,L, 1.12 mmol) was added to a solution of4-
benzyloxycarbonylamino-1-hydroxybutylidene-1,1-bisphosphonic acid (144 mg,
0.38 mmol) in 1 mL dry DMF, followed by the dropwise addition of a solution of
the
iodomethyl ester (520 mg, 0.814 mmol) from step (d) in 3 mL DMF. After
stirring at
ambient temperature for 5.5 h, the reaction mixture was concentrated on a
rotavapor,
2o treated with EtOAc (20 mL), and filtered. The precipitates were washed with
10 mL
more EtOAc. The organic solution was washed with 10 mL saturated aqueous
NaHC03 containing a small amount of NazS203, followed by reextraction of the
aqueous phase with 10 mL EtOAc. The organic phases were combined, washed with
mL brine, dried over anhydrous NazS04, and concentrated. Flash column
25 chromatography (silica gel, 5 - 25 % MeOH in CHZClZ (20 mL) yielded a
fraction
that gave light yellow solids (59 mg) enriched in the diester. The solids were
dissolved in 25 mL EtOAc, washed with 5% aqueous EDTA-disodium salt (2 x 2
mL), dried over anhydrous Na2S04, and evaporated to dryness under vacuum.
30 'H NMR (250 MHz, CDCI, + 1% CD30D) 8 0.81-0.97 (m, 12H), 1.60-2.50 (br,
l OH), 3.09 (br s, 2H), 3.55-3.77 (m, 4H), 4.20 (br s, 2H), 4.43 (m, 2H), 4.80-
5.40 (m,
12H), 5.64 (br, 4H), 7.28-7.35 (m, 25H);
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3'P NMR (101 MHz, CDC13 + 1% CD30D) 8 18.3.
PGT/SE99/00528
b) 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid di[traps-4-(N
benzyloxycarbonyl-L-valyloxy)-L-prolyoxymethyl] ester.
The product of step a) is deCBz protected by gentle conditions as described in
Greene supra to yield the title compound.
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Biological Example 1
135
The bioavailability of prodrugs of the invention built on the bis-phosphonate
alendronate was assayed in rats. The prodrugs of the invention were orally
administered at a dose corresponding to 0.1 mmol/kg to triplicate rats in a
propylene
glycol vehicle. The mother compound was also administered IV at 1/10 of that
does
for the calculation of absolute oral bioavailability.
Urine was collected over 24 hours in a metabolic cage and analysed as shown in
to Kline et al J Chromat. 534 (1990) 139-149, but modified as follows: 1 ml of
urine is
mixed with 50 p,l or 1.25 M calcium chloride and 100 p.l of 1 M sodium
hydroxide.
After centrifugation, the urine was aspirated off and the pellet redissolved
in 0.8 ml
0.2 M acetic acid, 0.4 ml of 0.01 M EDTA and 0.4 ml of 0.2 m sodium acetate. 1
ml
of water was added and the solution loaded onto a preconditioned DEA
cartridge.
i5 The cartridge was washed with 1 ml of water and alendronate eluted with 1
ml of 1M
carbonate buffer, pH 10.4. A part of the eluent, 150 ~l was mixed with 5 p,l
of
0.05 M potassium cyanide and 5 p,l of NDA solution (1 mg/ml) in methanol. 50
p,l
was injected into the chromatograph.
2o Absolute bioavailability is determined as the ratio between the AUC for the
mother
compound administered N against the AUC for the prodrug administered orally.
Results are presented in table 1 below:
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Table 1
136
compound absolute bioavailability (%)
alendronate 2.2
Example A-2 42
Example A-11 34
Example A-3 25
Example A-S 14
Example A-4 12
Example A-18 10
Example A-10
Example A-19 g
Example A-24 11
Example A-22 g
Example A-23 10
Example A-12 g
The compounds of the invention exhibit substantially enhanced oral
bioavailability
relative to the alendronate. The pivaloyloxymethyl alendronate ester presented
as the
preferred prodrug in US 5 227 506 provided no better bioavailability than the
mother
compound in this assay (results not shown).
Although the invention has been illustrated by reference to a number of
examples
1 o built on the phosphorous-containing drug alendronate and employing various
linkers
and amino acids, it will be appreciated that the invention is not so limited
to these
values but extends throughout the spirit and scope of the accompanying claims.
