Title: D-proline prodrugs
Abstract: The invention relates to compounds of formulas ##STR1##
wherein
R1 and R2 are independently from each other lower alkoxy, lower alkenyloxy, benzyloxy, hydroxy, —OCH(CH3)OC(O)-lower alkyl or —OCH2C(O)N(R3)(R4), with the proviso that only one of R1 or R2 may be hydroxy; R3 and R4 are independently from each other and signify hydrogen, lower alkyl, lower alkenyl or cycloalkyl; or R1 and R2 form together with the carbon atom, to which they are attached the linking group X, wherein X is —O(CH2)nCH═CH(CH2)nO— or —O(CH2)mO—; n is 1, 2 or 3; and m is 4-8, or a pharmaceutically acceptable salt of said compound. Compounds of the present invention can be used for the treatment of diseases where Serum Amyloid P Component depletion has a beneficial effect, in particular in the treatment or prevention of central and systemic amyloidosis.
Patent Number: 6,903,129 Issued on 06/07/2005 to Huwyler, et al.
| Inventors:
|
Huwyler; Joerg (Burg, CH);
Jakob-Roetne; Roland (Inzlingen, DE);
Poli; Sonia Maria (Basle, CH)
|
| Assignee:
|
Hoffman-La Roche Inc. (Nutley, NJ)
|
| Appl. No.:
|
307699 |
| Filed:
|
December 2, 2002 |
Foreign Application Priority Data
| Current U.S. Class: |
514/411; 514/422; 540/455; 548/524 |
| Intern'l Class: |
C07D 267/22; C07D207/00; C07D281/18; A61K031/40 |
| Field of Search: |
548/524
540/455
514/411,422
|
References Cited [Referenced By]
U.S. Patent Documents
| 5691336 | Nov., 1997 | Dorn et al.
| |
| 6103910 | Aug., 2000 | Hertel et al.
| |
| Foreign Patent Documents |
| 915 088 | May., 1999 | EP.
| |
Other References
Bungaard, H., Drugs of the Future, 16, pp. 443-458 (1991).
Houston, J. B., Biochem. Pharmacol., 47, pp. 1469-1479 (1994).
Noursadeghi et al., Proc. Natl. Acad. Sci. USA, 97, pp. 14584-14589 (2000).
Anderson F.M., et al., Anti-Cancer Drug Design, vol. 15, pp. 119-126 (2000).
|
Primary Examiner: Kifle; Bruck
Attorney, Agent or Firm: Johnston; George W., Rocha-Tramaloni; Patricia S.
Claims
1. A compound of formula
##STR5##
wherein
R
1 and R
2 are independently from each other lower alkoxy,
lower alkenyloxy, hydroxy, —OCH(CH
3)OC(O)-lower alkyl or —OCH
2C(O)N(R
3)(R
4),
with the proviso that only one of R
1 or R
2 may be hydroxy;
R
3 and R
4 are independently from each other hydrogen, lower
alkyl, lower alkenyl or cycloalkyl; or
R
1 and R
2 form together with the carbon atom, to which
they are attached the linking group X, wherein
X is —O(CH
2)
nCH═CH(CH
2)
nO—
or —O(CH
2)
mO—;
n is 1, 2 or 3; and
m is 4-8;
or a pharmaceutically acceptable salt thereof.
2. The compound of formula I in accordance with claim 1, wherein R
1 and
R
2 are identical.
3. The compound of formula I in accordance with claim 2, wherein R
1 and
R
2 are both —OCH
2C(O)N(R
3)(R
4)
and R
3 and R
4 are independently from each other hydrogen,
lower alkyl, lower alkenyl or cycloalkyl.
4. The compound of formula I in accordance with claim 3, wherein the compound is
(R)-1-[6-[(R)-2-carbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid carbamoylmethyl ester,
(R)-1-[6-[(R)-2-allylcarbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid allylcarbamoylmethyl ester,
(R)-1-{6-[(R)-2-(isopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid isopropylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(tert-butylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(cyclopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid cyclopropylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(dimethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid dimethylcarbamoyl-methyl ester or
(R)-1-{6-[(R)-2-(diethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diethylcarbamoyl-methyl ester.
5. The compound of formula I in accordance with claim 2, wherein R
1 and
R
2 are both lower alkoxy.
6. The compound of formula I in accordance with claim 5, wherein the compound is
(R)-1-{6-[(R)-2-methoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid methyl ester,
(R)-1-{6-[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid ethyl ester or
(R)-1-{6-oxo-6-[(R)-2-propoxycarbonyl-pyrrolidin-1-yl]-hexanoyl}-pyrrolidine-2-carboxylic
acid propyl ester.
7. A compound of formula
##STR6##
wherein X is —O(CH
2)
nCH═(CH
2)
nO—
or —O(CH
2)
mO—;
n is 1, 2 or 3; and
m is 4 to 8;
or a pharmaceutically acceptable salt thereof.
8. The compound in accordance with claim 7, wherein X is —O(CH
2)
nCH═CH(CH
2)
nO—.
9. The compound in accordance with claim 8, wherein the compound is
(12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracos-16-ene-2,7,13,20-tetraone,
(12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacos-17-ene-2,7,13,22-tetraone
or
(12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacos-18-ene-2,7,13,24-tetraone.
10. The compound of formula IA in accordance with claim 7, wherein X is —O(CH
2)
mO—.
11. The compound of formula IA in accordance with claim 10, wherein the compound is
(12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[9.3.0.0 8,12]tetracosane-2,7,13,20-tetraone,
(12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacosane-2,7,13,22-tetraone
or
(12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacosane-2,7,13,24-tetraone.
12. A pharmaceutical composition comprising one or more compounds of formula I
##STR7##
wherein
R
1 and R
2 are independently from each other lower alkoxy,
lower alkenyloxy, hydroxy, —OCH(CH
3)OC(O)-lower alkyl or —OCH
2C(O)N(R
3)(R
4),
with the proviso that only one of R
1 or R
2 may be hydroxy;
R
3and R
4 are independently from each other hydrogen, lower
alkyl, lower alkenyl or cycloalkyl; or
R
1 and R
2 form together with the carbon atom, to which
they are attached the linking group X, wherein
X is —O(CH
2)
nCH═CH(CH
2)
nO—
or —O(CH
2)
mO—;
n is 1, 2 or 3; and
m is 4-8
and a pharmaceutically acceptable excipient.
13. A pharmaceutical composition comprising one or more compounds of formula
IA
##STR8##
wherein X is —O(CH
2)
nCH═(CH
2)
nO—
or —O(CH
2)
mO—;
n is 1, 2 or 3; and
m is 4 to 8
and a pharmaceutically acceptable excipient.
Description
FIELD OF THE INVENTION
The present invention is concerned with new D-prolines of formulas
##STR2##
wherein
R1 and R2 are independently from each other lower
alkoxy, lower alkenyloxy, benzyloxy, hydroxy, —OCH(CH3)OC(O)-lower
alkyl or —OCH2C(O)N(R3)(R4), with the proviso
that only one of R1 or R2 may be hydroxy;
R3 and R4 are independently from each other hydrogen,
lower alkyl, lower alkenyl or cycloalkyl; or
R1 and R2 form together with the carbon atom, to which
they are attached the linking group X, wherein
X is —O(CH2)nCH═CH(CH2)nO—
or —O(CH2)mO—;
n is 1, 2 or 3; and
m is 4-8,
as well as pharmaceutically acceptable salts of said compounds.
BACKGROUND OF THE INVENTION
The D-proline of formula II (parent compound) is a known compound and is disclosed
in EP 915 088. Formula II is provided as
##STR3##
Compounds of formula II have a limited bioavailability. It was therefore
useful to find derivatives of the compound of formula II to render these compounds
suitable for oral application.
A molecule with optimal structural configuration and physicochemical properties
for eliciting the desired therapeutic response at its target site does not necessarily
possess the best molecular form and properties for delivery to its point of ultimate
action. Usually, only a minor fraction of doses administered reach the target area
and since most agents interact with non-target sites as well, an inefficient delivery
may result in undesirable side effects. This fact of differences in transport and
in situ effect characteristics for many drug moleculs is the fundamental reason
why bioreversible chemical derivatization of drugs, i.e, prodrug formation, is
a means by which a substantial improvement in the overall efficacy of drugs can
be achieved.
Therefore, the prodrug approach involves
1. enhancement of bioavailability and passage through various biological barriers,
2. increased duration of pharmacological effects,
3. increased site-specificity,
4. decreased toxicity and adverse reactions,
5. improvement of organoleptic properties, and
6. improvement of stability and solubility.
A prodrug is a pharmacologically inactive derivative of a parent drug molecule
that requires spontaneous or enzymatic transformation within the body in order
to release the active drug, and that has improved delivery properties over the
parent drug molecule. Prodrugs are designed to overcome pharmaceutically and/or
pharmacokinetically based problems associated with the parent drug molecule that
would otherwise limit the clinical usefulness of the drug.
In recent years several types of bioreversible derivatives have been exploited
for designing prodrugs. Using esters as a prodrug type for drugs containing carboxyl
or hydroxyl function is most popular. Further well-known are prodrug derivatives
of peptides, 4-imidazolidinones and the like, described in
Drugs of the Future,
1991, 16(5), 443-458 or N-oxides, described, for example, in U.S. Pat. No. 5,691,336.
It is desirable to provide novel compounds of formulas I and IA to overcome pharmaceutically
and/or pharmacokinetically based problems associated with the parent drug molecule
of formula II that would otherwise limit the clinical usefulness of the drug.
SUMMARY OF THE INVENTION
An aspect of the present invention is directed to a compound of formula I or
IA,
and their pharmaceutically acceptable salt.
Another embodiment of this invention is directed to a compound of formula
I or IA for use as prodrug in the treatment or prevention of central and systemic amyloidosis.
Another embodiment of the present invention is directed to a pharmaceutical
composition comprising one or more compounds of formula I and IA and a pharmaceutically
acceptable excipient, particularly those suitable for the treatment of diseases
related to central and systemic amyloidosis.
Yet another embodiment of this invention is directed to a method of treating
diseases related to central and systemic amyloidosis comprising administering to
a patient in need of such treatment a therapeutically effective amount of at least
one compound of formula I or IA.
PREFERRED DESCRIPTION OF THE INVENTION
Compounds of the present invention can be used for the treatment of diseases
where Serum Amyloid P Component depletion has a beneficial effect, in particular
in the treatment or prevention of all forms of central and systemic amyloidosis.
The most common disorders associated with amyloidosis are Alzheimer's disease,
maturity onset diabetes mellitus or amyloidosis
- as a significant cause of non-ischaemic heart failure,
- as complication of long term haemodialysis in renal failure,
- as complication of monoclonal gammopathies,
- from chronic inflammatory disorders,
- from chronic infections,
- or from certain types of cancer.
Furthermore, amyloidosis comprises many different diseases such as forms
of hereditary amyloidosis, and most commonly familial amyloid polyneuropathy (FAP),
scrapie and Kreuzfeld-Jakob disease.
The compounds of the present invention may also be used in certain bacterial
infections (M. Noursadeghi et. al., Proc. Natl.Acad. Sci. USA 97 (2000) 14584-14589).
It has now surprisingly been found that compounds of formulas I and IA were,
in
vitro and in vivo, readily converted to the parent compound of formula II
##STR4##
and can therefore be used as prodrugs.
The most preferred embodiment of this invention is the prodrugs of formula I.
Exemplarly preferred are compounds of formula I, wherein R
1 and R
2
are identical, R
1 and R
2 are —OCH
2C(O)N(R
3)(R
4)
or lower alkoxy, and R
3 and R
4 are independently from each
other hydrogen, lower alkyl, lower alkenyl or cycloalkyl.
Preferred compounds, wherein R
1 and R
2 are —OCH
2C(O)N(R
3)(R
4),
are the followings:
(R)-1-[6-[(R)-2-carbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid carbamoylmethyl ester,
(R)-1-[6-[(R)-2-allylcarbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid allylcarbamoylmethyl ester,
(R)-1-{6-[(R)-2-(isopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid isopropylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(tert-butylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(cyclopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid cyclopropylcarbamoyl-methyl ester,
(R)-1-{6-[(R)-2-(dimethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid dimethylcarbamoyl-methyl ester or
(R)-1-{6-[(R)-2-(diethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diethylcarbamoyl-methyl ester.
Preferred compounds, wherein R
1 and R
2 are lower
alkoxy, are the followings:
(R)-1-{6-[(R)-2-methoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid methyl ester,
(R)-1-{6-[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid ethyl ester or
(R)-1-{6-oxo-6-[(R)-2-propoxycarbonyl-pyrrolidin-1-yl]-hexanoyl}-pyrrolidine-2-carboxylic
acid propyl ester.
The invention relates further to compounds of formula IA, wherein X is
Compounds, wherein X is —O(CH
2)
nCH═CH(CH
2)
nO—,
are the followings:
(12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracos-16-ene-2,7,13,20-tetraone,
(12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacos-17-ene-2,7,13,22-tetraone or
(12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacos-18-ene-2,7,13,24-tetraone.
Compounds, wherein X is —O(CH
2)
mO—
are for example the followings:
(12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracosane-2,7,13,20-tetraone,
(12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacosane-2,7,13,22-tetraone or
(12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacosane-2,7,13,24-tetraone.
As used herein "pharmaceutically acceptable salts" useful in this invention include
salts derived from metals, salts from amino acids and salts of mineral or organic
acids. Examples of preferred metal salts are those derived from the alkali metals,
for example, lithium (Li
+), sodium (Na
+) and potassium (K
+).
Especially preferred is sodium. Other salts are derived from amino acids such as,
for example, salts with arginine or lysine.
In the formulas represented herein, when substituents are illustrated as joined
to the nucleus a solid line () indicates that the substituent is in the α-orientation,
that is, below the plane of the molecule.
The term "lower alkyl" refers to both straight and branched chain saturated hydrocarbon
groups having 1 to 6 and preferably 1 to 4 carbon atoms, for example, methyl, ethyl,
n-propyl, isopropyl and tertiary butyl.
By the term "cycloalkyl" is meant a 3-6 membered saturated carbocyclic moiety,
e.g., cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, in particular cyclopentyl.
The compound of formula I or a salt thereof can be produced by methods known
in the art. Moreover, a compound of formula I can be prepared by esterifying the
compound of formula II or a salt thereof with a compound of formulas
wherein Y is a halogen atom, preferably a chlorine atom; R
5 is
lower alkyl, lower alkenyl, benzyl, —CH(CH
3)OC(O)-lower alkyl
or —CH
2C(O)N(R
3)(R
4); and R
3 and
R
4 are described above.
In the esterification reaction, the starting compound III is used in a proportion
of about 1 to 3 mole equivalents to each equivalent of the starting compound II
or a salt thereof.
Examples of compounds of formula III are the followings: 2-chloroacetamide,
N-(chloroacetyl)allylamine, N(chloroacetyl)isopropylamine, N-(chloroacetyl)-t-butyl-amine,
N-(chloroacetyl)-cyclopropylamine, 2-chloro-N,N-dimethylacetamide, 2-chloro-N,N-diethylacetamide,
2-chloro-N,N-diisopropylacetamide or 2-chloro-N-t-butyl-N-methylacetamide.
This reaction is carried out in a solvent inert to the reaction. Suitable solvents
include N,N-dimethylforamide, N,N-dimethylacetamide, acetone, and acetonitrile.
Examples 1-9 have been prepared in this way.
The compound of formula I, wherein R
1 and R
2 are both methoxy
(Example 10) may be prepared by a reaction of a Solution of diazomethane in diethylether
with a solution of the compound of formula II in tetrahydrofuran.
Furthermore, compounds of Examples 11-16 have been prepared by reactions
of a solution of a compound of formula II and Amberlite® IR120 (ion-exchange
resin, useful in catalytic applications) in conventional manner with ethanol, propanol,
butanol, allylalcohol, 3-buten-1-ol and 4-penten-1-ol.
The compound of formula I, wherein R
1 is ethoxy and R
2 is
benzyloxy (Example 17) has been prepared from a mixture of adipic acid anhydride,
D-proline-O-benzyl hydrochloride and N-methyl-morpholine in dichloromethane with
a polymer bound primary amine and with a mixture of N-methyl-morpholin, 1-hydroxybenzotriazole,
1-(3-dimethylethylaminopropyl)-3-ethylcarbodiimide hydrochloride and H-D-Proline-O-ethyl.
The benzyloxy group may then be hydrogenated to the hydroxy group in the presence
of palladium/carbon in ethylacetate (Example 18).
A compound of formula I, wherein R
1 and R
2 are both —OCH(CH
3)OC(O)-t-butyl
(Example 19) maybe prepared from a solution of a compound of formula II with diazabicycloundecan
with 2,2-dimethyl-propionic acid (RS)-1-bromo-ethyl ester at room temperature.
Further, a compound of formula IA may be prepared by reaction of (R)-1-{6-[(R)-2-alkoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid allyl ester (Example 20), or (R)-1-{-[(R)-2-but-3-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid but-3-enyl ester (Example 21), or (R)-1-{6-[(R)-2-pent-4-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid pent-4-enyl ester (Example 22) with benzylidene-bis(tricyclohexylphosphine)-dichlororuthenium
in dichloromethane. The reaction is carried out at about 50° C.
The double bond in compounds of Examples 20, 21 and 22 may then be hydrogenated
with palladium/carbon in ethylacetate in conventional manner to compounds of Examples
23, 24 and 25.
As mentioned earlier, the compounds of formula I and their pharmaceutically usable
addition salts may be used as prodrugs of the parent compounds of formula II, which
possess valuable pharmacological properties.
These compounds were investigated in accordance with the test provided below.
The evidence, that the compounds of formula I may be used as prodrugs of their
parent compounds of formula II is shown in accordance with the description given hereinafter.
The conversion of prodrugs to the corresponding parent compounds is via a hydrolytic
mechanism and there is well known evidence from the literature that similar reactions
occur in vivo.
Test Description
Stability of the Prodrugs in Blood and Plasma Samples
Plasma and blood samples from different species were merged with equimolar
amounts (10 μM) of prodrug and parent drug in DMSO and incubated for different
time intervals (up to 60 min.) at 37° C. The reaction was stopped by protein
precipitation with acetonitrile followed by centrifugation (20 min., 1800 g at
10° C.). The supernatant was immediately subjected to analysis.
The concentration of formed parent compound was determined by LC-MS. The chromatographic
system was comprised of a trapping column (X-Terra™ MS C8 3.5 μm,
10×2.1 mm i.d., Waters) and an analytical column (Symmetry C8 3.5 μm,
50×2.1 mm i.d., Waters) connected to a SCIEX API 2000 triplequadrupole mass
spectrometer equipped with a turbo ion spray interface. The mobile phases were
1% aqueous formic acid and acetonitrile. The parent compound together with its
deuterium labelled internal standard was enriched on the trapping column and eluted
with a fast gradient. The retention time of (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid was ˜2.1 min. The effluent (300 μl min
-1) was passed
to the turbo ion spray interface without splitting and was nebulized using nitrogen.
Multiple reaction monitoring (MRM) in positive mode Was used for mass spectrometric
detection. The transitions for the parent were 341.1 [M+H]
+ to 226.1
[Fragment]
+ and for the internal standard 349.1 [M+H]
+ to
234.1 [Fragment]
+. The results were expressed as half-lifes (50% conversion
of the prodrug), using the data of the prodrug at time-point 0 min. as 0% value.
Test Description for Microsome Incubation
Rat and human liver microsome incubations were conducted on-line in a CTC PAL
autosampler in order to avoid any degradation of the prodrugs during the work-up.
Incubation mixtures consisted of liver microsomes (rat 1.0 mg prot/mL or human
2.0 mg prot/mL), prodrug 10 μM, MgCl
2 (3.3 mM), and an NADPH regenerating
system consisting of glucose-6-phosphate dehydrogenase, NADPH and glucose-6-phosphate
equivalent to 1 mM NADPH) in a total volume of 1.0 mL of potassium phosphate buffer
100 mM pH 7.4.
Reactions were initiated by addition of the NADPH regenerating system at
37° C. At time 1, 5, 9, 13, 17, 21, 25, and 29 min a 5 μL aliquot was
directly analysed on a HPLC-MS/MS system comprising of a HP 1100 quaternary pump
with degasser and a PE-Sciex API-2000 MS/MS spectrometer. The analytical column
was a Waters Symmetry Shield RP8 (2.1*50 mm with a 3.5 μM particle size).
A polarity non linear gradient from phase A (MeOH/Ac. Form.1% 20/80) to phase B
(MeOH) was applied for a total run time of 2 minutes at a flow rate of 0.25 mL/min.
The PE-Sciex API-2000 MS/MS spectrometer was used for detection of both the prodrugs
and the parent compound (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid. In vivo metabolic clearance was predicted according to published procedures
(Houston, J. B., Biochem. Pharmacol., 47:1469-1479, 1994). In brief, the intrinsic
clearance (Clearance, Table 1) was calculated from the measured in vitro half-life
taking into account incubation volume and microsomal protein used for the in vitro
incubation. The intrinsic clearance was expressed in terms of ul/min/mg microsomal
protein. For in vivo extrapolations, the hepatic extraction ratio (E) was calculated.
Reported below is the %MAB value (maximal achievable bioavailability), which is
equal to 1-E.
Results
| TABLE 1 |
| Example |
Rat microsomes |
Human microsomes |
Rat plasma |
| No. |
Clearance |
MAB % |
Clearance |
MAB % |
tl/2 hours |
| 1 |
5 |
88 |
1 |
93 |
0.4 |
| 2 |
18 |
66 |
64 |
20 |
low |
| 3 |
82 |
30 |
16 |
50 |
low |
| 4 |
625 |
5 |
57 |
22 |
low |
| 5 |
36 |
50 |
8 |
67 |
low |
| 6 |
7 |
84 |
1 |
96 |
0.8 |
| 7 |
9 |
73 |
2 |
90 |
0.2 |
| 10 |
9 |
79 |
2 |
90 |
1.3 |
| 11 |
46 |
44 |
5 |
77 |
0.4 |
| 12 |
253 |
12 |
73 |
18 |
Table 1 shows that the compounds of formula I are potential prodrugs for the
parent compound II.
It has been found, that the compounds of the formula I exhibit low stability
in
plasma where they give rise to the formation of the parent compound II. With microsomes
they show a medium to low stability with the formation of compound II. The bioavailability
was measured for selected examples: 12 (100%), 3 (8%), 5 (8%) and 13 (10%). For
comparison, parent compound of formula II: 4%.
These findings suggest that the compounds of formula I show an increased oral
bioavailability and therefore have potential value for the treatment of diseases
where SAP depletion has a beneficial effect in particular as described above.
In accordance to the tests, the compounds of formula I can function as prodrugs
of their parent compounds of formula II.
The compounds of formula I as well as their pharmaceutically usable acid addition
salts can be used as medicaments or pharmaceutical composition, e.g., in the form
of pharmaceutical preparations. The pharmaceutical preparations can be administered
orally, e.g., in the form of tablets, coated tablets, dragées, hard and soft
gelatine capsules, solutions, emulsions or suspensions. The administration can,
however, also be effected rectally, e.g., in the form of suppositories, or parenterally,
e.g., in the form of injection solutions.
The compounds of formula I and their pharmaceutically usable acid addition salts
can be processed with pharmaceutically inert, inorganic or organic excipients for
the production of tablets, coated tablets, dragees and hard gelatine capsules.
Lactose, corn starch or derivatives thereof, talc, stearic acid or its salts can
be used as such excipients e.g., for tablets, dragées and hard gelatine capsules.
Suitable excipients for soft gelatine capsules include vegetable oils, waxes,
fats, semi-solid and liquid polyols.
Suitable excipients for the manufacture of solutions and syrups include
water, polyols, saccharose, invert sugar and glucose.
Suitable excipients for injection solutions include water, alcohols, polyols,
glycerol and vegetable oils.
Suitable excipients for suppositories are natural or hardened oils, waxes,
fats, semi-liquid and liquid polyols.
Moreover, the pharmaceutical preparations can contain preservatives, solubilizers,
stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts
for varying the osmotic pressure, buffers, masking agents or antioxidants. They
can also contain still other therapeutically valuable substances.
The dosage can vary within wide limits and will, of course, be fitted to the
individual requirements in each particular case. In general, in the case of oral
administration a daily dosage of about 10 to 1000 mg per person of a compound of
general formula I should be appropriate, although the above upper limit can also
be exceeded when necessary.
The following Examples 1 to 25 illustrate the present invention without limiting
it. All temperatures are given in decrees Celsius.
The following prodrugs have been prepared:
EXAMPLE 1
(R)-1-[6-[(R)-2-Carbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid carbamoylmethyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 93.5 mg (1 mmol) 2-chloroacetamide in 2 ml dimethylformamide were added
14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine. After stirring
overnight at 90° C. the solvent was distilled off, the residue was taken up
with dichloromethane and extracted with water, 2% aqueous sodium bicarbonate and
brine. The organic extracts were dried with sodium sulfate and the solvent was
distilled off to yield 100 mg (44%) of (R)-1-[6-[(R)-2-carbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid carbamoylmethyl ester as a light yellow foam, MS m/e (%): 455 (M+H+, 100).
EXAMPLE 2
(R)-1-[6-[(R)-2-Allylcarbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid allylcarbamoylmethyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 134 mg (1 mmol) N-(chloroacetyl)allylamine in 3 ml dimethylformamide were
added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine. After
stirring, overnight at 90° C. the solvent was distilled off, the residue was
taken up with dichloromethane and extracted with water. The organic extracts were
dried with sodium sulfate and the solvent was distilled off to yield 200 mg (75%)
of (R)-1-[6-[(R)-2-allylcarbamoylmethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid allylcarbamoylmethyl ester as a yellow oil, MS m/e (%): 535 (M+H+, 100).
EXAMPLE 3
(R)-1-{6-[(R)-2-(Isopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid isopropylcarbamoyl-methyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 136 mg (1 mmol) N-(chloroacetyl)isopropyl-amine in 3 ml dimethylformamide
were added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine.
After stirring overnight at 90° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with water. The organic extracts
were dried with sodium sulfate and the solvent was distilled off to yield 200 mg
(74%) of (R)-1-{6-[(R)-2-(isopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid isopropylcarbamoyl-methyl ester as a yellow solid, MS m/e (%): 539 (M+H+, 100).
EXAMPLE 4
(R)-1-{6-[(R)-2-(tert-Butylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylcarbamoyl-methyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 149 mg (1 mmol) N-(chloroacetyl)-t-butyl-amine in 3 ml dimethylformamide
were added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine.
After stirring overnight at 90° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with water. The organic extracts
were dried with sodium sulfate and the solvent was distilled off to yield 205 mg
(72%) of (R)-1-{6-[(R)-2-(tert-butylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylcarbamoyl-methyl ester as a white solid, MS m/e (%): 567 (M+H+, 100).
EXAMPLE 5
(R)-1-{6-[(R)-2-(Cyclopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid cyclopropylcarbamoyl-methyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 134 mg (1 mmol) N-(chloroacetyl)-cyclopropyl-amine in 3 ml dimethylformamide
were added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine.
After stirring overnight at 90° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with water. The organic extracts
were dried with sodium sulfate and the solvent was distilled off to yield 190 mg
(71%) of (R)-1-{6-[(R)-2-(cyclopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid cyclopropylcarbamoyl-methyl ester as a light yellow solid, MS m/e (%): 535
(M+H+, 100).
EXAMPLE 6
(R)-1-{6-[(R)-2-(Dimethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid dimethylcarbamoyl-methyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 103 ml (1 mmol) 2-chloro-N,N-dimethylacetamide in 2.5 ml dimethylformamide
were added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine.
After stirring overnight at 100° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with water. The organic extracts
were dried with sodium sulfite and the solvent was distilled off to yield 190 mg
(84%) of (R)-1-{6-[(R)-2-(dimethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid dimethylcarbamoyl-methyl ester as a yellow oil, MS m/e (%): 511 (M+H+, 100).
EXAMPLE 7
(R)-1-{6-[(R)-2-(Diethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diethylcarbamoyl-methyl ester
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 137 ml (1 mmol) 2-chloro-N,N-diethylacetamide in 3 ml dimethylformamide
were added 14.9 mg (0.1 mmol) sodium iodide and 139 ml (1 mmol) triethylamine.
After stirring overnight at 90° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with water. The organic extracts
were dried with sodium sulfate and the solvent was distilled off to yield 260 mg
(92%) of (R)-1-{6-[(R)-2-(diethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diethylcarbamoyl-methyl ester as a yellow oil, MS m/e (%): 567 (M+H+, 100).
EXAMPLE 8
(R)-1-{6-[(R)-2-(Diisopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diisopropylcarbamoyl-methyl ester
To a solution of 680 mg (2.0 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 708 mg (4 mmol) 2-chloro-N,N-diisopropylacetamide in 10 ml dimethylformamide
were added 60.0 mg (0.4 mmol) sodium iodide and 557 ml (4 mmol) triethylamine.
After stirring overnight at 90° C. the solvent was distilled off, the residue
was taken up with dichloromethane and extracted with 2% aqueous sodium bicarbonate
and brine. The organic extracts were dried with sodium sulfate and the solvent
was distilled off to yield 885 mg (71%) of (R)-1-{6-[(R)-2-(diisopropylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid diisopropylcarbamoyl-methyl ester as an oil, MS m/e (%): 623 (M+H+, 100).
EXAMPLE 9
(R)-1-{6-[(R)-2-(tert-Butylmethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylmethylcarbamoyl-methyl ester
To a solution of 680 mg (2.0 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 652 mg (4 mmol) 2-chloro-N-t-butyl-N-methylacetamide in 10 ml dimethylformamide
were added 60.0 mg (0.4 mmol) sodium iodide and 557 ml (4 mmol) triethylamine.
After stirring over the weekend at 90° C. the solvent was distilled off, the
residue was taken up with dichloromethane and extracted with 2% aqueous sodium
bicarbonate and brine. The organic extracts were dried with sodium sulfate and
the solvent was distilled off to yield 1.02 g (85% ) of (R)-1-{6-[(R)-2-(tert-butylmethylcarbamoyl-methoxycarbonyl)-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid tert-butylmethylcarbamoyl-methyl ester as a solid, MS m/e (%): 595 (M+H+, 100).
EXAMPLE 10
(R)-1-{6-[(R)-2-Methoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid methyl ester
A solution of diazomethane in diethylether (˜6 mmol) was added to a solution
of 500 mg (1.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid in 10 ml tetrahydrofuran. After stirring overnight methanol was added and
the solvents were evaporated. The residue was taken up in dichloromethane, extracted
with brine and dried with sodium sulfate. After separation from the solvent 400
mg (72%) (R)-1-{6-[(R) -2-methoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid methyl ester were obtained as a light yellow oil, MS m/e (%): 369 (M+H+, 100).
EXAMPLE 11
(R)-1-{6-[(R)-2-Ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid ethyl ester
A mixture of 170 mg, (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 54 mg Amberlite® IR120 in 10 ml ethanol were stirred at room temperature
for 48 hours. After filtration the solvent was distilled off, the residue was taken
up in dichloromethane and extracted with 2% aqueous sodium bicarbonate. The organic
extract was dried with sodium sulfate and the solvent was distilled off to yield
105 mg (53%) (R)-1-{6-[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid ethyl ester as alight yellow oil, MS m/e (%): 397 (M+H+, 100).
EXAMPLE 12
(R)-1-{6-Oxo-6-[(R)-2-propoxycarbonyl-pyrrolidin-1-yl]-hexanoyl}-pyrrolidine-2-carboxylic
acid propyl ester
A mixture of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 54 mg Amberlite® IR120 in 10 ml propanol were stirred at room temperature
for 48 hours. After filtration the solvent was distilled off, the residue was taken
up in dichloromethane and extracted with 2% aqueous sodium bicarbonate. The organic
extract was dried with sodium sulfate and the solvent was distilled off to yield
65 mg (31%) (R)-1-{6-oxo-6-[(R)-2-propoxycarbonyl-pyrrolidin-1-yl]-hexanoyl}-pyrrolidine-2-carboxylic
acid propyl ester as alight yellow oil, MS m/e (%): 424 (M+, 3), 268 (100), 156
(21), 70 (69).
EXAMPLE 13
(R)-1-{6-[(R)-2-Butoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid butyl ester
A mixture of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 54 mg Amberlite® IR120 in 10 ml butanol were stirred at room temperature
for 48 hours. After filtration the solvent was distilled off, the residue was taken
up in dichloroethane and extracted with 2% aqueous sodium bicarbonate. The organic
extract was dried with sodium sulfate and the solvent was distilled off to yield
70 mg (31%) (R)-1-{6-[(R)-2-butoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid butyl ester as alight yellow oil, MS m/e (%): 453 (M+H+, 100).
EXAMPLE 14
(R)-1-{6-[(R)-2-Alloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid allyl ester
A mixture of 1.02 g (3 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 3 g Amberlite® IR120 in 30 ml allylalcohol were stirred at room temperature
for 90 hours. After filtration the solvent was distilled off, the residue was taken
up in dichloromethane and extracted with 2% aqueous sodium bicarbonate. The organic
extract was dried with sodium sulfate and the solvent was distilled off to yield
620 mg (49%) (R)-1-{6-[(R)-2-alloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid allyl ester as alight yellow oil, MS m/e (%): 421 (M+H+, 100).
EXAMPLE 15
(R)-1-[6-[(R)-2-But-3-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidin-2-carboxylic
acid but-3-enyl ester
A mixture of 1.02 g (3 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 3 g Amberlite® IR120 in 25 ml 3-buten-1-ol were stirred at room temperature
for 72 hours. After filtration the solvent was distilled off, the residue was taken
up in dichloromethane and extracted with 2% aqueous sodium bicarbonate. The organic
extract was dried with sodium sulfate and the solvent was distilled off to yield
490 mg (37%) (R)-1-[6-[(R)-2-but-3-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid but-3-enyl ester as alight yellow oil, MS m/e (%): 449 (M+H+, 100).
EXAMPLE 16
(R)-1-[6-[(R)-2-Pent-4-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid pent-4-enyl ester
A mixture of 2.04 g (6 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 5 g Amberlite® IR120 in 25 ml 4-penten-1-ol were stirred at room
temperature for 48 hours. After filtration the solvent was distilled off, the residue
was taken up in dichloromethane and extracted with 2% aqueous sodium bicarbonate.
The organic extract was dried with sodium sulfate and the solvent was distilled
off to yield 1.6 g (58%) (R)-1-[6-[(R)-2-pent-4-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid pent-4-enyl ester as a light yellow oil, MS m/e (%): 477 (M+H+, 100).
EXAMPLE 17
(R)-1-[6-[(R)-2-Ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid benzyl ester
A mixture of 403 mg (3.15 mmol) adipic acid anhydride, 604 mg (2.5 mmol) D-proline-benzyl-ester
hydrochloride and 274 ml (2.5 mmol) N-methyl-morpholin in 10 ml dichloromethane
was stirred at room temperature for 1 hour. Then 0.5 g polymer bound primary amine
(2 meq/g) were added and stirring was continued for 1 hour. After filtration 823
ml (7.5 mmol) N-methyl-morpholin, 338 mg (2.5 mmol) 1-hydroxybenzotriazole, 479
mg (2.5 mmol) 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 450
mg (2.5 mmol) D-proline-ethyl-ester were added and the mixture was stirred for
another 48 hours. Removal of the solvent and chromatography on silicagel with ethylacetate
gave 135 mg (12%) (R)-1-[6-[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid benzyl ester as a light yellow oil, MS m/e (%): 459 (M+H+, 100).
EXAMPLE 18
(R)-1-[6-[(R)-2-Ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic acid
A mixture of 92 mg (0.2 mmol) (R)-1-[6-[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid benzyl ester and 30 mg 10% palladium/carbon in 5 ml ethylacetate was hydrogenated
over night. Filtration and removal of the solvent yielded 70 mg (R)-1-[6[(R)-2-ethoxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid as a light yellow oil, MS m/e (%): 369 (M+H+, 100).
EXAMPLE 19
(R)-1-(6-{(R)-2-[1-(2,2-Dimethyl-propionyloxy)-ethoxycarbonyl]-pyrrolidin-1-yl}-6-oxo-hexanoyl)-pyrrolidine-2-carboxylic
acid 1-(2,2-dimethyl-propionyloxy)-ethyl ester (mixture of diastereomers)
To a solution of 170 mg (0.5 mmol) (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid and 149 ml (1 mmol) diazabicycloundecan in 4 ml dimethylformamide were added
209 mg (1 mmol) 2,2-dimethyl-propionic acid (RS)-1-bromo-ethyl ester and the mixture
was stirred at room temperature for 3 hours. The solvent was distilled off and
the residue was taken up in water and extracted with dichloromethane. The organic
extracts were washed with 2% aqueous sodium bicarbonate and buffer pH7 and dried
with sodium sulfate. Removal of the solvent and chromatography on silicagel with
ethylacetate gave 55 mg (18%) (R)-1-(6-{(R)-2-[1-(2,2-dimethyl-propionyloxy)-ethoxycarbonyl]-pyrrolidin-1-yl}-6-oxo-hexanoyl)-pyrrolidine-2-carboxylic
acid 1-(2,2-dimethyl-propionyloxy)-ethyl ester (mixture of diastereomers) as a
colorless oil, MS m/e (%): 597 (M+H+, 100).
EXAMPLE 20
(12R,21R)-14,19-Dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracos-16-ene-2,7,13,20-tetraone
A mixture of 420 mg (1 mmol) (R)-1-{6[-(R)-2-alloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl}-pyrrolidine-2-carboxylic
acid allyl ester and 40 mg benzylidene-bis(tricyclohexylphosphine)dichlororuthenium
in 30 ml dry dichloromethane was stirred at 50° C. overnight. Removal of the
solvent and chromatography on silicagel with ethylacetate/acetone with a ratio
of 8/2 gave 110 mg (28%) (12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracos-16-ene-2,7,13,20-tetraone
as light yellow oil, MS m/e (%): 393 (M+H+, 100).
EXAMPLE 21
(12R,23R)-14,21-Dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacos-17-ene-2,7,13,22-tetraone
A mixture of 410 mg (0.92 mmol) (R)-1-[6-[(R)-2-but-3-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid but-3-enyl ester and 40 mg benzylidene-bis(tricyclohexylphosphine)dichlororuthenium
in 30 ml dry dichloromethane was stirred at 50° C. overnight. Removal of the
solvent and chromatography on silicagel with ethylacetate/acetone with a ratio
of 8/2 gave 200 mg (51%) (12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacos-17-ene-2,7,13,22-tetraone
as an oil, MS m/e (%): 421 (M+H+, 100).
EXAMPLE 22
(12R,25R)-14,23-Dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacos-18-ene-2,7,13,24-tetraone
A mixture of 1.20 g (2.5 mmol) (R)-1-[6-[(R)-2-pent-4-enyloxycarbonyl-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic
acid pent-4-enyl ester and 40 mg benzylidene-bis(tricyclohexylphosphine)dichlororuthenium
in 30 ml dry dichloromethane was stirred at 50° C. overnight. Removal of the
solvent and chromatography on silicagel with ethylacetate/acetone with a ratio
of 8/2 gave 430 mg (38%) (12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacos-18-ene-2,7,13,24-tetraone
as an oil, MS m/e (%): 449 (M+H+, 100).
EXAMPLE 23
(12R,21R)-14,19-Dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracosane-2,7,13,20-tetraone
A mixture of 260 mg (0.66 mmol) (12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0
8,12]tetracos-16-ene-2,7,13,20-tetraone and 30 mg 10% palladium/carbon in 10 ml
ethylacetate was hydrogenated over night. Filtration and removal of the solvent
yielded 255 mg (98%) (12R,21R)-14,19-dioxa-1,8-diaza-tricyclo[19.3.0.0 8,12]tetracosane-2,7,13,20-tetraone
as a light yellow oil, MS m/e (%): 395 (M+H+, 100).
EXAMPLE 24
(12R,23R)-14,21-Dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacosane-2,7,13,22-tetraone
A mixture of 150 mg (0.36 mmol) (12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0
8,12]hexacos-17-ene-2,7,13,22-tetraone and 20 mg 10%) palladium/carbon in 5 ml
ethylacetate was hydrogenated over night. Filtration and removal of the solvent
yielded 120 mg (79%) (12R,23R)-14,21-dioxa-1,8-diaza-tricyclo[21.3.0.0 8,12]hexacosane-2,7,13,22-tetraone
as a light yellow oil, MS m/e (%): 423 (M+H+, 100).
EXAMPLE 25
(12R,25R)-14,23-Dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacosane-2,7,13,24-tetraone
A mixture of 400 mg (0.9 mmol) (12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0
8,12]octacos-18-ene-2,7,13,24-tetraone and 40 mg 10% palladium/carbon in 10 ml
ethylacetate was hydrogenated over night. Filtration and removal of the solvent
yielded 245 mg (61%) (12R,25R)-14,23-dioxa-1,8-diaza-tricyclo[23.3.0.0 8,12]octacosane-2,7,13,24-tetraone
as a white solid, MS m/e (%): 451 (M+H+, 100).
EXAMPLE A
| Tablets of the following composition are manufactured: |
| |
Active substance |
5 |
| |
Lactose |
45 |
| |
Corn starch |
15 |
| |
Microcrystalline cellulose |
34 |
| |
Magnesium stearate |
1 |
| |
Tablet weight |
100 |
| |
EXAMPLE B
| Capsules of the following composition are manufactured: |
| |
Active substance |
10 |
| |
Lactose |
155 |
| |
Corn starch |
30 |
| |
Talc |
5 |
| |
Capsule fill weight |
200 |
| |
The active substance, lactose and corn starch are firstly mixed in a mixer and
then in a comminuting machine. The mixture is returned to the mixer, the talc is
added thereto and mixed thoroughly. The mixture is filled by machine into hard
gelatine capsules.
EXAMPLE C
Suppositories of the following composition are manufactured:
| |
Active substance |
15 |
| |
Suppository mass |
1285 |
| |
Total |
1300 |
| |
The suppository mass is melted in a glass or steel vessel, mixed thoroughly and
cooled to 45° C. Thereupon, the finely powdered active substance is added
thereto and stirred until it has dispersed completely. The mixture is poured into
suppository molds of suitable size, left to cool, the suppositories are then removed
from the molds and packed individually in wax paper or metal foil.
EXAMPLE D
An injection solution may have the following composition and is manufactured:
| |
| |
Active substance |
1.0 |
mg |
| |
1 n HCl |
20.0 |
μl |
| |
acetic acid |
0.5 |
mg |
| |
NaCl |
8.0 |
mg |
| |
phenol |
10.0 |
mg |
| |
1 n NaOH |
q.s. ad pH 5 |
| |
H2O |
q.s. ad 1 |
ml |
| |
*
