SYNTHESIS OF DGJNAc PROM D-GLUCURONOLACTONE AND USE
TO INHIBIT alpha-N-ACETYLGALACTOSAMINIDASES
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Patent Application
No. 611282,393, filed February 2,2010, the entire disclosures of which are incorporated by
reference.
FIELD
[0002] The present disclosure relates in general to the use and synthesis of iminosugars for
medical purposes and, in particular, to the use of iminosugars for inhibiting a-Nacetylgalactosaminidases
(GalNAcases) or P-hexosaminidases, as well as treatments for
diseases associated with these enzymes.
SUMMARY
[0003] According to one embodiment, the current invention discloses a method for
synthesizing DGJNAc or a DGJNAc derivative from D-glucuronolactone. Synthesis of
DGJNAc comprises introducing nitrogen at C5 of glucuronolactone, inversion of
configuration of the hydroxyl group at C3 and formation of the piperidine ring by
introduction of nitrogen between C6 and C2.
[0004] According to another embodiment, numerous novel DGJNAc derivatives are
disclosed. These compositions include a compound of the formula,,
R 3
or a pharmaceutically acceptable salt thereof, wherein R is selected from the group consisting
of substituted or unsubstituted alkyl groups, substituted or unsubstituted cycloalkyl groups,
substituted or unsubstituted aryl groups, and substituted or unsubstituted oxaalkyl groups; or
wherein R is
R1 is a substituted or unsubstituted alkyl group;
XI-5 are independently selected from H, N02, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than carbonyl; and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and provided
that when Z is NH, Y is a substituted or unsubstituted C -alkyl group, other than carbonyl.
[0005] In another embodiment, the current invention is drawn to a method of inhibiting a-
N-acetylgalactosaminidases (GalNAcases) or P-hexosaminidases, comprising addition of a
compound of the formula,
or a pharmaceutically acceptable salt thereof, wherein R is selected from the group consisting
of hydrogen, substituted or unsubstituted alkyl groups, substituted or unsubstituted cycloalkyl
groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted oxaalkyl
groups; or wherein R is
R1 is a substituted or unsubstituted alkyl group;
are independently selected from H, NO2, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than carbonyl; and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and provided
that when Z is NH, Y is a substituted or unsubstituted C1-alkyl group, other than carbonyl,
to a composition comprising a-N-acetylgalactosaminidases (GalNAcases) or Phexosaminidases.
In a further embodiment, R is hydrogen and the compound is DGJNAc.
[0006] In another embodiment, the current invention is drawn to a method of treating or
preventing a disease associated with a-N-acetylgalactosaminidases (GalNAcases) or Phexosaminidases
activity comprising: administering to a subject in need thereof an effective
amount of a compound of the formula,
R 3
or a pharmaceutically acceptable salt thereof, wherein R is selected from the group consisting
of hydrogen, substituted or unsubstituted alkyl groups, substituted or unsubstituted cycloalkyl
groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted oxaalkyl
groups; or wherein R is
R1 is a substituted or unsubstituted alkyl group;
are independently selected from H, NO2, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than carbonyl; and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and provided
that when Z is NH, Y is a substituted or unsubstituted C1-alkyl group, other than carbonyl,
[0007] In a further embodiment, the subject is a human being and the disease is Schindler
disease. In addition, R may be hydrogen and the compound DGJNAc
DETAILED DESCRIPTION
I. Iminosugars
[0008] Iminosugars, compounds in which the ring pyranose or furanose oxygen has been
replaced by nitrogen, are the archetype for interaction with carbohydrate processing enzymes.
(1). However, among the myriad of sugar mimics reported, there is not a single example of
efficient inhibition of a-N-acetyl-galactosaminidases (GalNAcases).
. .
11. Overview
[0009] The current invention reports DGJNAc 1D and its derivatives as the first potent,
specific and competitive inhibitors of GalNAcases. In addition, D-glucuronolactone 2D, a
well-established chiron for the synthesis of many homochiral targets including amino acids
(2) and iminosugars, (3) can be used as the starting material for an efficient synthesis of
DGJNAc [2-acetamido-1 ,2-dideoxy-D-galacto-nojirimycin] 1D in an overall yield of 20%.
The L-enantiomers of many iminosugars have surprising biological activities compared to
their D-natural products. (4) The synthesis of L-DGJNAc 2L from the readily available (5) Lglucuronolactone
2L is also provided. The only previous synthesis of 1D starts from
deoxynojirimycin (6) and a racemic mixture of 1D and 1L has also been prepared. (7)
However, no investigations of the glycosidase inhibitory properties of 1D have previously
been identified.
. . .
111. Synthesis of DGJNAc from glucuronolactone
[0010] The synthesis of DGJNAclD requires introduction of nitrogen at C5 of
glucuronolactone (Scheme I), inversion of configuration of the hydroxyl group at C3 and
formation of the piperidine ring by introduction of nitrogen between C6 and C2 (with
inversion of configuration).
1 D 2D 2L 1 L 3
[0011] Scheme 1: Strategy for synthesis of DGJNAc 1 enantiomers - numbering of C in
DGJNAc derived from that in glucuronolactone 2
H q OH
H AcHN
P h H N ~ O
4 5
0
6
Figure: Potent P-hexosaminidase inhibitors
iv. Schindler Disease
[0012] Schindler disease, a congenital metabolic disorder, is a lysosomal storage disorder
caused by a deficiency in the alpha-NAGA (alpha-N-acetylgalactosaminidase) enzyme. This
lysosomal storage disorder is also known as Kanzaki disease and Alpha-Nacetylgalactosaminidase
deficiency. Mutations to the NAGA gene on chromosome 22 lead to
a build up of glycoproteins in the lysosomes and an accumulation of glycosphingolipids
throughout the body. This accumulation of sugars causes the clinical features associated with
this disease. Schindler disease is an autonomic recessive disorder.
[0013] There are three main types of the disease. In the Type I infantile form, babies
develop normally until about a year old. Afterwards, the child begins to lose previously
acquired skills associated with the coordination of physical and mental behaviors. Additional
neurological and neuromuscular symptoms including diminished muscle tone, weakness,
involuntary rapid eye movements, vision loss, and seizures may be present. Over time
symptoms worsen and children experience a decreased ability to move certain muscles due to
muscle rigidity and the ability to respond to external stimuli decreases. Other symptoms
include neuroaxonal dystrophy from birth, discoloration of skin, Telangiectasia or widening
of blood vessels.
[0014] In Type I1 adult form, symptoms are milder and may not appear until the mid 30s.
Angiokeratomas, an increased coarsening of facial features, and mild intellectual impairment
are typical symptoms. Type I11 form is considered an intermediate disorder with varying
symptoms among patients. Severe symptoms include seizures and mental retardation. Less
severe symptoms include delayed speech, mild autistic like presentation, and/or behavioral
problems.
[0015] Other lysosomal enzymes are also well known for their role in numerous diseases.
One example of these enzymes are P-hexosaminidases. Selective inhibition of Phexosaminidases
are useful for the study of osteoarthritis, (8) allergy, (9) Alzheimer's
disease, (10) 0-GlcNAcase inhibition, (1 1) cancer metastasis, (12) type I1 diabetes, (13)
genetic diseases such as Tay-Sachs and Sandhoff diseases, (14) and of plant regulation. (15)
The synthetic piperidine analogue of N-acetylglucosamine DGJNAc 3 (16) and its N-alkyl
derivatives (17) are potent inhibitors of P-hexosaminidases. The natural product nagstatin 4,
(1 8) with a galacto-configuration, does not inhibit GalNAcases even though it is a potent
inhibitor of P-hexosaminidases. (19) The synthetic analogue with a gluco-configuration 5
(20) together with PUG derivatives 6 (21) and GlcNAc-thiazolines 7 (22) are very potent
inhibitors of P-hexosaminidases. A rare example of a pyrrolidine potent hexosaminidase
inhibitor is LABNAc 8; (23) the first pyrrolizidine P-hexosaminidase inhibitor, pochonicine 9
[or its enantiomer], has been isolated from a fungal strain Pochonia suchlasporia var.
suchlasporia TAMA 87. (24) Some seven membered-ring imino sugars also display potent
inhibition. (25)
[0016] Unless otherwise specified "a" or "an" means one or more.
vi. DGJNAc and its DGJNAc derivatives
[0017] The present inventors discovered that certain iminosugars, such as DGJNAc and
DGJNAc derivatives, may be effective in the inhibition of GalNAcases or Phexosaminidases.
In particular, the DGJNAc and DGJNAc derivatives may be useful for
treating or preventing a disease or condition caused by or associated with GalNAcases) or Phexosaminidases.
[0018] In many embodiments, the iminosugar is DGJNAc or a DGJNAC derivative.
DGJNAc (2-acetamido- 1, 2-dideoxy-D-galacto-nojirimycini)s a compound of the formula
H
[0019] A "DGJNAc derivative" is a derivative of DGJNAc wherein the ring nitrogen is not
substituted with a hydrogen atom.
[0020] In general, DGJNAc and DGJNAc derivatives can be represented by the formula
R
[0021] wherein R may be selected from hydrogen (for DGJNAc), substituted or
unsubstituted alkyl groups, substituted or unsubstituted cycloalkyl groups, substituted or
unsubstituted aryl groups, or substituted or unsubstituted oxaalkyl groups.
[0022] In some embodiments, R may be substituted or unsubstituted alkyl groups and/or
substituted or unsubstituted oxaalkyl groups comprise from 1 to 16 carbon atoms, from 4 to
12 carbon atoms or from 8 to 10 carbon atoms. The term "oxaalkyl" refers to an alkyl
derivative, which may contain from 1 to 5 or from 1 to 3 or from 1 to 2 oxygen atoms. The
term "oxaalkyl" includes hydroxyterminated and methoxyterrninated alkyl derivatives.
[0023] In some embodiments, R may be selected from, but is not limited to -(CH2)60CH3,
-(CH2)60CH2CH3, -(CH2)60(CH2)2CH3, -(CH2)60(CH2)3CH3, -(CH2)20(CH2)5CH3,
-(CH2)20(CH2)6CH3;-(CH2)20(CH2)7CH-(3C; H2)yOH; -(CH2)90CH3.
[0024] In some embodiments, R may be an branched or unbranched, substituted or
unsubstituted alkyl group, which may contain up 20 carbon atoms. In some embodiments,
the alkyl group may C2-C12 or C3-C7 alkyl group.
[0025] In certain embodiments, the alkyl group may be a long chain alkyl group, which
may be C6-C20 alkyl group; C8-C16 alkyl group; or C8-C10 alkyl group. In some
embodiments, R may be a long chain oxaalkyl group, i.e., a long chain alkyl group, which
may contain from 1 to 5 or from 1 to 3 or from 1 to 2 oxygen atoms.
[0026] In some embodiments, R may have the following formula
where R1 is a substituted or unsubstituted alkyl group;
XI-5 are independently selected from H, N02, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than carbonyl; and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and provided
that when Z is NH, Y is a substituted or unsubstituted C1-alkyl group, other than carbonyl.
[0027] In some embodiments, Z is NH and R1-Y is a substituted or unsubstituted alkyl
group, such as C2-C20 alkyl group or C4-C12 alkyl group or C4-C10 alkyl group.
[0028] In some embodiments, X1 is NO2 and X3 is N3. In some embodiments, each of X2,
Xq and X5 is hydrogen.
vii. Salts, Prodrugs, Pharmaceutical Compositions
[0029] In some embodiments, the iminosugar may be in a form of a salt derived from an
inorganic or organic acid. Pharmaceutically acceptable salts and methods for preparing salt
forms are disclosed, for example, in Berge et al. (J. Pharm. Sci. 66:l-18, 1977). Examples of
appropriate salts include but are not limited to the following salts: acetate, adipate, alginate,
citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate,
camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate,
glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate,
hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate,
persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate,
tosylate, mesylate, and undecanoate.
[0030] In some embodiments, the iminosugar may also used in a form of a prodrug.
[0031] In some embodiments, the iminosugar may be used as a part of a composition,
which further comprises a pharmaceutically acceptable carrier and1 or a component useful for
delivering the composition to an animal. Numerous pharmaceutically acceptable carriers
useful for delivering the compositions to a human and components useful for delivering the
composition to other animals such as cattle are known in the art. Addition of such carriers
and components to the composition of the invention is well within the level of ordinary skill
in the art.
[0032] In some embodiments, the pharmaceutical composition may consist essentially of
DGJNAc or the DGJNAc derivative, indicating that the DGJNAc or the DGJNAc derivative
is the only active ingredient in the composition.
[0033] Yet in some other embodiments, DGJNAc or the DGJNAc derivative may be
administered with one or more additional compounds.
[0034] In some embodiments, the iminosugar may be used in a liposome composition, such
as those disclosed in US publication 200810 13835 1; US application No. 1214 10,750 filed
March 25,2009 and US provisional application No. 611202,699 filed March 27,2009.
viii. Administration and Inhibition of GalNAcases or Phexosaminidases
[0035] The iminosugar, such as a DGJNAc or the DGJNAc derivative, may be
administered to a cell or an animal affected by disorders associated with GalNAcases or Phexosaminidases
activity. The iminosugar may inhibit GalNAcases or P-hexosaminidases
and help reduce, abate, or diminish the disease in the animal.
[0036] In addition, DGJNAc or the DGJNAc derivative, may be used to study the inhibition
of GalNAcases or P-hexosaminidases with in vitro or in vivo studies.
[0037] Animals suffering from the disease include primates including monkeys and
humans.
[0038] The amount of iminosugar administered to an animal or to an animal cell to the
methods of the invention can be an amount effective to inhibit the GalNAcases or Phexosaminidases.
The term "inhibit" as used herein may refer to the detectable reduction
and/or elimination of a biological activity exhibited in the absence of the iminosugar. The
term "effective amount" may refer to that amount of the iminosugar necessary to achieve the
indicated effect. The term "treatment" as used herein may refer to reducing or alleviating
symptoms in a subject, preventing symptoms from worsening or progressing, inhibition or
elimination of the causative agent, or prevention of the disorder related to the GalNAcases or
P-hexosaminidases activity in a subject.
[0039] The amount of the iminosugar which may be administered to the cell or animal is
preferably an amount that does not induce toxic effects which outweigh the advantages which
accompany its administration.
[0040] Actual dosage levels of active ingredients in the pharmaceutical compositions may
vary so as to administer an amount of the active compound(s) that is effective to achieve the
desired therapeutic response for a particular patient.
[0041] The selected dose level may depend on the activity of the iminosugar, the route of
administration, the severity of the condition being treated, and the condition and prior
medical history of the patient being treated. However, it is within the skill of the art to start
doses of the compound(s) at levels lower than required to achieve the desired therapeutic
effect and to gradually increase the dosage until the desired effect is achieved. If desired, the
effective daily dose may be divided into multiple doses for purposes of administration, for
example, two to four doses per day. It will be understood, however, that the specific dose
level for any particular patient can depend on a variety of factors, including the body weight,
general health, diet, time and route of administration and combination with other therapeutic
agents and the severity of the condition or disease being treated. The adult human daily
dosage may range from between about one microgram to about one gram, or from between
about 10 mg and 100 mg, of the iminosugar per 10 kilogram body weight. Of course, the
amount of the iminosugar which should be administered to a cell or animal may depend upon
numerous factors well understood by one of skill in the art, such as the molecular weight of
the iminosugar and the route of administration.
[0042] Pharmaceutical compositions that are useful in the methods of the invention may be
administered systemically in oral solid formulations, ophthalmic, suppository, aerosol, topical
or other similar formulations. For example, it may be in the physical form of a powder,
tablet, capsule, lozenge, gel, solution, suspension, syrup, or the like. In addition to the active
agent, such pharmaceutical compositions may contain pharmaceutically-acceptable carriers
and other ingredients known to enhance and facilitate drug administration. Other possible
formulations, such as nanoparticles, liposomes resealed erythrocytes, and immunologically
based systems may also be used to administer the agent. Such pharmaceutical compositions
may be administered by a number of routes. The term "parenteral" used herein includes
subcutaneous, intravenous, intraarterial, intrathecal, and injection and infusion techniques,
without limitation. By way of example, the pharmaceutical compositions may be
administered orally, topically, parenterally, systemically, or by a pulmonary route.
[0043] These compositions may be administered in a single dose or in multiple doses which
are administered at different times.
[0044] The present invention can be illustrated in more details by the following example,
however, it should be understood that the present invention is not limited thereto.
[0045] Embodiments described herein are further illustrated by, though in no way limited
to, the following working examples.
EXAMPLE
Example 1
Synthesis of DJGNAc 1D from D-glucuronolactone 2D
C 12 R=H 14 R=H 16 R=H
(iii)
13 R = SiMedBu 15 R=Bn 17 R = OSOzCF3
[Sil= SiMe2Bu Bn = CH,ph AC = CH,CO (viii)
t
OAc (xi) ~3%@:~O~A- c -
H Bn Bn
OMe
Me0
Scheme 2: (i) (CF3S02)20, CH2C12, pyridine; then NaN;, DMF, 99% (ii) DIBALH, CH2C12,
pyridine; then NaBH4, H20, 72% (iii) ' B U M ~ ~ S ~pyCri~di,n e, 99% (iv) PCC, CH2C12,
molecular sieve; then NaBH4, EtOH, H20, 79% (v) PhCH2Br, NaH, DMF, 97% (vi) MeOH,
HC1, 97% (vii) (CF3S02)20, CH2C12, pyridine (viii) PhCH2NH2, THF, (ix) Et20:BF;, Ac20,
93% (x) DIBALH, CH2C12, pyridine; then NaBH4, H20, 83% (xi) Zn, THF/AcOH/Ac20,
79% (xii) MeONa, MeOH; then H2, Pd (10%C), HC1, dioxane, H20, 98%.
[0046] For the synthesis of DJGNAc 1D from D-glucuronolactone 2D, the acetonide 10
was esterified with trifluoromethanesulfonic (triflic) anhydride in dichloromethane in the
presence of pyridine and the resulting crude triflate was treated with sodium azide in DMF to
give the ido-azide 11 {mp. 1 12-1 14 "C; [alDZ+52 61.4 (c 1. O, CHCI;) [lit. (29) mp. 11 4-11 6
"C , [alDZ+O24 3 (c 1.1, CHCl;)]} in 97% yield. Direct conversion of the azidolactone 11 by
a number of hydrides to the diol 12 gave only low yields; such a-azidolactones are extremely
sensitive to base and commonly a two step reduction is necessary with initial reduction to the
lactol. Accordingly DIBALH reduction of the azidolactone 11 in dichloromethane gave the
corresponding lactol which was further reduced by sodium borohydride in methanol to afford
the diol 12 mp. 120-122 "C, [alDZ-659 .6 (c 0.94, CHCl;) in 72% yield. Selective protection
of the primary alcohol in 12 by reaction with tert-butyldimethylsilyl (TBDMS) chloride gave
the corresponding TBDMS ether 13, oil, [alDZ-152 .7 (c 1.1, CHCl;) in 99% yield; the overall
yield of 13 from glucuronolactone 2D was 72% on a multigram scale and without any need
for chromatographic purification until the final stage.
[0047] The synthesis of DGJNAc 1D required inversion and subsequent protection of the
remaining unprotected C3 OH in the silyl ether 13. Oxidation of 13 with pyridinium
chlorochromate in dichloromethane in the presence of molecular sieve afforded the
corresponding ketone which on reduction from the least hindered face of the carbonyl gave
the inverted alcohol 14, oil, [a]D2+57 4.9 (c 0.94, CHC13), in 79% yield. Treatment of 14 with
benzyl bromide and sodium hydride in DMF formed the fully protected benzyl ether 15, oil,
[a]D2+51 01.5 (c 0.56, CHC13) in 97% yield. Both the acetonide and silyl protecting groups in
15 were removed by treatment with HC1 in methanol to give a 5: 1 mixture of anomers of the
methyl furanoside 16 (97%); reaction of 16 with triflic anhydride in dichloromethane in the
presence of pyridine gave the ditriflate 17 which, with benzylamine in THF, gave the bicyclic
pyrrolidine 18, oil, +22.8 (c 1.11, CHC13), as a single anomer in an overall yield of
6 1 %. Formation of a piperidine ring by cyclization of a ditriflate was thus efficient; examples
of successful cyclizations of a ditriflate, such as the formation of a pyrrolidine, (30) are very
rare.
[0048] Acetolysis of the furanoside 18 with boron trifluoride etherate in acetic anhydride
gave a 4: 1 mixture of the epimers 19 in 93% yield. The OMe group in 19 was reductively
removed by sequential treatment with DIBALH in dichloromethane followed by sodium
borohydride in methanol; acetylation of the resulting diol allowed easy isolation of the
diacetate 20, oil, [a]D2+579 .8 (c 0.43, CHC13), in 83% overall yield from 19. Rapid reduction
of the azide in XK by zinc dust in the presence of copper(I1) sulfate in acetic acid-acetic
anhydride-THF (3 1) with subsequent acylation of the corresponding amine gave the
crystalline triacetate 21, mp. 112-1 14 "C, [alDZ+526 .2 (c 1.1, Me2CO) in 79% yield.
Removal of the acetate protecting groups by treatment of 21 with sodium methoxide in
methanol followed by hydrogenolysis of the benzyl groups by palladium (10% on carbon) in
dioxane: aqueous hydrochloric acid gave DGJNAc lD,mp. 150-154 "C, [a]D2+54 1.9 (c 0.67,
H20) ) [lit6o il, [[aD2+03 7 (c 1, MeOH)], in 98% yield. Unlike many iminosugars, the free
base DGJNAc is readily crystallized; the overall yield of DGJNAc 1D from Dglucuronolactone
2D was 20%.
[0049] Selected data for DGJNAc ID: HRMS (ESI +ve): C8Hl6N2NaOf4o und 227.1001;
(M+N~+r)e quires 227.1002; +41.9 (c 0.67, H20); m.p. 150-154°C; vmax( thin film, Ge): 3287
(br, s, OH/NH), 1637 (s, amide I), 1561 (s, amide 11); 6" (D20,400 MHz): 2.00 (3H, s, Me),
2.37 (lH, dd, Hla Jge1m2. 9, Jla,2 11.6), 2.76 (lH, dt, H5 J5,4 1.3,J5,6a=J5,6b6 .6), 3.08 (lH,
dd, Hlb Jge1m2. 9,Jlb,25 .1), 3.58 (lH, dd, H3 J3,2 10.6, J3,4 3.0), 3.61 (lH, dd, H6a Jge1m1. 1,
J6a6,.53) , 3.65 (lH, dd, H6b Jge1m1. 1, J6b6,.56) , 3.96 (lH, dt, H2 J2,i1a1 .1, J2,i5b.1 , J2,3
11 .I), 4.01 (lH, dd, H4 J4,3 3.0, J4,5 1.4); &C (D20, 100 MHz): 22.7 (Me), 47.7 (Cl), 49.1
(C2), 59.4 (C5), 61.9 (C6), 68.9 (C4), 73.2 (C3), 175.2 (GOMe); LRMS (ESI +ve): 205
(77%, M+H+), 43 1 (loo%, 2 ~ + ~ a + ) .
Example 2
Synthesis of L-DJGNAc 1L from L-glucuronolactone 2L
[0050] The enantiomer L-DGJNAc lL, mp. 152-156 'C, [a]D2456 .6 (c 0.73, H20),w as
prepared by an identical procedure from L-glucuronolactone 2L.
Example 3
Inhibition of GalNAcases and P-hexosaminidases by DJGNAc 1D and L-DGJNAc 1L
[0051] DGJNAc 1D was a highly potent competitive inhibitor of GalNAcases (Ki 0.081 pM
from chicken liver, Ki 0.136 pM from Charonia lampas); DGJNAc 1D was a good but much
less potent competitive inhibitor of P-hexosaminidases (IC50 1.8 pM from Jack bean, IC50 1.8
pM from Jack bean, ICso 4.2 pM from bovine kidney, ICso 8.3 pM from human placenta,
IC5o 2.2 pM from HL-60).
[0052] The enantiomer L-DGJNAc lL, showed no inhibition of a-Nacetylgalactosaminidases
but was a very weak but non-competitive inhibitor of Phexosaminidases
[Ki 1100 pM - compared with Ki 2.2 pM for DGJNAc 1D - from human
placenta]. This result was in accord with Asano's hypothesis (32) that L-enantiomers show
non-competitive inhibition whereas D-imino sugars usually are competitive inhibitors.
[0053] DGJNAc 1D showed modest inhibition of coffee bean a-galactosidase (IC5o 64 pM)
whereas L-DGJNAc lL, showed no inhibition of this enzyme.
[0054] Both enantiomers of DGJNAc 1 were screened as inhibitors of a number of other
glycosidases and neither enantiomer showed any significant inhibition [less than 50%
inhibition at 1000 mM) against a-glucosidases (rice, yeast), P-glucosidases (almond, bovine
liver), P-galactosidase (bovine liver), a-mannosidase (Jack bean), P-glucuronidases (E. coli,
bovine liver), a-L-rharnnosidase (P. decumbens), or a-L-fucosidase (bovine epididymis).
[0055] Although the foregoing refers to particular preferred embodiments, it will be
understood that the present invention is not so limited. It will occur to those of ordinary skill
in the art that various modifications may be made to the disclosed embodiments and that such
modifications are intended to be within the scope of the present invention.
[0056] All of the publications, patent applications and patents cited in this specification are
incorporated herein by reference in their entirety.
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Asymmetry 2000, 11, 1645-1680. (d) Watson, A. A.; Fleet, G. W. J.; Asano, N.; Molyneux,
R. J.; Nash, R. J. Phytochemistry 2001,56,265-295.
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WE CLAIM:
1. A method for synthesizing DGJNAc or a DGJNAc derivative from Dglucuronolactone,
comprising
introducing nitrogen at C5 of glucuronolactone5 ,
inversion of the configuration of the hydroxyl group at C3, and
formation of the piperidine ring by introduction of nitrogen between C6 and C2.
2. A compound of the following formula,
N
R
OH
AcHN OH
CH3 OH
,
10 or a pharmaceutically acceptable salt thereof, wherein R is selected from the
group consisting of substituted or unsubstituted alkyl groups, substituted or unsubstituted
cycloalkyl groups, substituted or unsubstituted aryl groups, and substituted or
unsubstituted oxaalkyl groups;
or wherein R is
Y Z
X1 X2
X3
X5 X4
R1
15
R1 is a substituted or unsubstituted alkyl group;
X1-5 are independently selected from H, NO2, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than
carbonyl; and
20 Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and
provided that when Z is NH, Y is a substituted or unsubstituted C1-alkyl group, other than
carbonyl.
3. A method of inhibiting a-N-acetylgalactosaminidases (GalNAcases) or Phexosaminidases,
comprising addition of a compound of the formula,
R ,
or a pharmaceutically acceptable salt thereof, wherein R is selected from the group
5 consisting of hydrogen, substituted or unsubstituted alkyl groups, substituted or unsubstituted
cycloalkyl groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted
oxaalkyl groups;
or wherein R is
10 R1 is a substituted or unsubstituted alkyl group;
XI-5 are independently selected from H, N02, N3, or NH2;
Y is absent or is a substituted or unsubstituted C1-alkyl group, other than carbonyl;
and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and
15 provided that when Z is NH, Y is a substituted or unsubstituted C1-alkyl group, other than
carbonyl,
to a composition comprising a-N-acetylgalactosaminidases (GalNAcases) or Phexosaminidases.
4. The method of claim 3, where R is hydrogen.
WO 2011/095893
22
PCT/IB2011/000380
5. A method of treating or preventing a disease associated with a-Nacetylgalactosaminidases
(GalNAcases) or P-hexosaminidases activity comprising:
administering to a subject in need thereof an effective amount of a compound of the
formula,
or a pharmaceutically acceptable salt thereof, wherein R is selected from the group
consisting of hydrogen, substituted or unsubstituted alkyl groups, substituted or unsubstituted
cycloalkyl groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted
oxaalkyl groups;
or wherein R is
Ri is a substituted or unsubstituted alkyl group;
Xi_5 are independently selected from H, NO2, N3, or NH2;
Y is absent or is a substituted or unsubstituted Ci-alkyl group, other than carbonyl;
and
Z is selected from a bond or NH; provided that when Z is a bond, Y is absent, and
provided that when Z is NH, Y is a substituted or unsubstituted Ci-alkyl group, other than
carbonyl,
6. The method of claim 5, wherein the subject is a human being.
7. The method of claim 5, wherein said subject has Schindler Disease.
WO 2011/095893 PCT/IB2011/000380
23
8. The method of claim 5, wherein R is hydrogen.