U.S. patent application number 09/788947 was filed with the patent office on 2001-08-16 for compounds useful for inhibition of farnesyl protein transferase.
Invention is credited to Alvarez, Carmen, Doll, Ronald J., Lalwani, Tarik, Liu, Yi-Tsung.
Application Number | 20010014681 09/788947 |
Document ID | / |
Family ID | 26727732 |
Filed Date | 2001-08-16 |
United States Patent
Application |
20010014681 |
Kind Code |
A1 |
Doll, Ronald J. ; et
al. |
August 16, 2001 |
Compounds useful for inhibition of farnesyl protein transferase
Abstract
Novel compounds of the formula: 1 are disclosed. In Formula 1.0
a represents N or NO, R.sup.1 and R.sup.3 are halo, R.sup.2 and
R.sup.4 are independently H or halo provided that at least one is
H, X is C, CH or N, and T represents a five or six membered
heterocycloalkyl ring having one or two heteroatoms selected from S
or O. Also disclosed are methods of inhibiting farnesyl protein
transferase and methods for treating tumor cells.
Inventors: |
Doll, Ronald J.; (Maplewood,
NJ) ; Alvarez, Carmen; (Roselle Park, NJ) ;
Lalwani, Tarik; (Edison, NJ) ; Liu, Yi-Tsung;
(Morris Township, NJ) |
Correspondence
Address: |
SCHERING-PLOUGH CORPORATION
PATENT DEPARTMENT (K-6-1, 1990)
2000 GALLOPING HILL ROAD
KENILWORTH
NJ
07033-0530
US
|
Family ID: |
26727732 |
Appl. No.: |
09/788947 |
Filed: |
February 20, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09788947 |
Feb 20, 2001 |
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09447491 |
Nov 23, 1999 |
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6228865 |
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60049951 |
Jun 17, 1997 |
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Current U.S.
Class: |
514/253.03 ;
514/290; 544/361; 546/93 |
Current CPC
Class: |
C07D 409/14 20130101;
C07D 221/16 20130101; C07D 405/14 20130101; A61P 35/00 20180101;
C07D 401/04 20130101 |
Class at
Publication: |
514/253.03 ;
514/290; 546/93; 544/361 |
International
Class: |
A61K 031/497; A61K
031/44; C07D 401/00 |
Claims
What is claimed is:
1. A compound of the formula: 215or a pharmaceutically acceptable
salt or solvate thereof, wherein: a represents N or NO.sup.--;
R.sup.1 and R.sup.3 are the same or different halo atom; R.sup.2
and R.sup.4 are selected from H and halo, provided that at least
one of R.sup.2 and R.sup.4 is H; the dotted line (---) represents
an optional bond; X is N, C when the optional bond is present, or
CH when the optional bond is absent; T is a substituent selected
from: 216wherein: A represents --(CH.sub.2).sub.b--; B represents
--(CH.sub.2).sub.d--; b and d are independently selected from: 0,
1, 2, 3, or 4 such that the sum of b and d is 3 or 4; and Y is
selected from: O, S, SO, or SO.sub.2; 217wherein: D represents
--(CH.sub.2).sub.e--; B represents --(CH.sub.2).sub.f--; e and f
are independently selected from: 0, 1, 2, or 3 such that the sum of
e and f is 2 or 3; and Z is O; 218wherein: F represents
--(CH.sub.2).sub.g--; G represents --(CH.sub.2).sub.h--; H
represents --(CH.sub.2).sub.i--; h represents 1, 2, or 3 g and i
are independently selected from: 0, 1 or 2 such that the sum of h,
g and i is 2 or 3; and V and W are independently selected from O,
S, SO, or SO.sub.2; 219wherein: the dotted line (---) represents an
optional bound; k is 1 or 2 such that when the optional bond is
present k represents 1, and when the optional double bond is absent
then k represents 2; R.sup.5, R.sup.6, R.sup.7 and R.sup.8 are the
same alkyl (preferably methyl); or R.sup.5 and R.sup.7 are the same
alkyl (preferably methyl), and R.sup.6 and R.sup.8 are H;
220wherein: the dotted lines (---) represent optional bonds 1 and 2
such that optional bonds 1 and 2 are both present, or optional
bonds 1 and 2 are both absent; Y represents O, S, SO, or SO.sub.2;
221wherein: Y represents O, S, SO, or SO.sub.2; 222wherein: R.sup.9
is selected from: --CN, --CO.sub.2H, or --C(O)N(R.sup.10).sub.2;
each R.sup.10 is the same or diferent alkyl group (preferably,
methyl); 223wherein: I represents --(CH.sub.2).sub.m--; m
represents 2 or 3; Y represents O, S, SO, or SO.sub.2; and R.sup.11
represents alkyl; 224
2. The compound of claim 1 having the formula: 225
3. The compound of claim 1 wherein R.sup.1 is halo, R.sup.2 is H,
R.sup.3 is halo, and R.sup.4 is H.
4. The compound of claim 3 wherein R.sup.1 is Br and R.sup.3 is
Cl.
5. The compound of claim 4 wherein X is CH, a is N, and the C5-C6
double bond is absent.
6. The compound of claim 1 wherein R.sup.1 is halo, R.sup.2 is
halo, R.sup.3 is halo, and R.sup.4 is H; or R.sup.1 is halo,
R.sup.2 is H, R.sup.3 is halo, and R.sup.4 is halo.
7. The compound of claim 6 wherein X is CH or N.
8. The compound of claim 7 wherein X is CH.
9. The compound of claim 7 wherein a is N, and the C5-C6 double
bond is absent.
10. The compound of claim 9 wherein R.sup.1 is Br, R.sup.2 is Br,
R.sup.3 is Cl, and R.sup.4 is H; or R.sup.1 is Br, R.sup.2 is H,
R.sup.3 is Cl, and R.sup.4 is Br.
11. The compound of claim 10 wherein X is CH.
12. The compound of claim 11 wherein R.sup.1 is Br, R.sup.2 is H,
R.sup.3 is Cl, and R.sup.4 is Br.
13. The compound of claim 12 having the formula: 226
14. The compound of claim 13 wherein T is 227
15. The compound of claim 14 wherein T is 228
16. The compound of claim 1 having the formula: 229wherein R.sup.12
is selected from: 230
15. The compound of claim 14 wherein R.sup.12 is 231
16. The compound of claim 1 selected from: 232
17. The compound of claim 1 having the formula: 233
18. A method of treating tumor cells expressing an activated ras
oncogene comprising administering an effective amount of a compound
of claim 1.
19. The method of claim 18 wherein the tumor cells treated are
pancreatic tumor cells, lung cancer cells, myeloid leukemia tumor
cells, thyroid follicular tumor cells, myelodysplastic tumor cells,
epidermal carcinoma tumor cells, bladder carcinoma tumor cells,
colon tumors cells, breast tumor cells and prostate tumor
cells.
20. A method of treating tumor cells wherein the Ras protein is
activated as a result of oncogenic mutation in genes other than the
Ras gene, comprising administering an effective amount of a
compound of claim 1.
21. A method of inhibiting farnesyl protein transferase comprising
the administration of an effective amount of the compound of claim
1.
22. A pharmaceutical composition for inhibiting farnesyl protein
transferase comprising an effective amount of compound of claim 1
in combination with a pharmaceutically acceptable carrier.
Description
BACKGROUND
[0001] WO 95/10516, published Apr. 20, 1995 discloses tricyclic
compounds useful for inhibiting farnesyl protein transferase.
[0002] In view of the current interest in inhibitors of farnesyl
protein transferase, a welcome contribution to the art would be
compounds useful for the inhibition of farnesyl protein
transferase. Such a contribution is provided by this invention.
SUMMARY OF THE INVENTION
[0003] This invention provides compounds useful for the inhibition
of farnesyl protein transferase (FPD. The compounds of this
invention are represented by the formula: 2
[0004] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0005] a represents N or NO.sup.--;
[0006] R.sup.1 and R.sup.3 are the same or different halo atom;
[0007] R.sup.2 and R.sup.4 are selected from H and halo, provided
that at least one of R.sup.2 and R.sup.4 is H;
[0008] the dotted line (---) represents an optional bond;
[0009] X is N, C when the optional bond is present, or CH when the
optional bond is absent;
[0010] T is a substituent selected from: 3
[0011] wherein:
[0012] A represents --(CH.sub.2).sub.b--;
[0013] B represents --(CH.sub.2).sub.d--;
[0014] b and d are independently selected from: 0, 1, 2, 3, or 4
such that the sum of b and d is 3 or 4; and
[0015] Y is selected from: O, S, SO, or SO.sub.2; 4
[0016] wherein:
[0017] D represents --(CH.sub.2).sub.e--;
[0018] B represents --(CH.sub.2).sub.f--:
[0019] e and f are independently selected from: 0, 1, 2, or 3 such
that the sum of e and f is 2 or 3; and
[0020] Z is O; 5
[0021] wherein:
[0022] F represents --(CH.sub.2).sub.g--;
[0023] G represents --(CH.sub.2).sub.h--;
[0024] H represents --(CH.sub.2).sub.i--;
[0025] h represents 1, 2, or 3
[0026] g and i are independently selected from: 0, 1 or 2 such that
the sum of h, g and i is 2 or 3; and
[0027] V and W are independently selected from O, S, SO, or
SO.sub.2; 6
[0028] wherein:
[0029] the dotted line (---) represents an optional bound:
[0030] k is 1 or 2 such that when the optional bond is present k
represents 1, and when the optional double bond is absent then k
represents 2;
[0031] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 are the same alkyl
(preferably methyl); or
[0032] R.sup.5 and R.sup.7 are the same alkyl (preferably methyl),
and R.sup.6 and R.sup.8 are H; 7
[0033] wherein:
[0034] the dotted lines (---) represent optional bonds 1 and 2 such
that optional bonds 1 and 2 are both present, or optional bonds 1
and 2 are both absent;
[0035] Y represents O, S, SO, or SO.sub.2; 8
[0036] wherein:
[0037] Y represents O, S, SO, or SO.sub.2; 9
[0038] wherein:
[0039] R.sup.9 is selected from: --CN, --CO.sub.2H, or
--C(O)N(R.sup.10).sub.2;
[0040] each R.sup.10 is the same or diferent alkyl group
(preferably, methyl); 10
[0041] wherein:
[0042] I represents --(CH.sub.2).sub.m--;
[0043] m represents 2 or 3;
[0044] Y represents O, S, SO, or SO.sub.2; and
[0045] R.sup.11 represents alkyl (preferably ethyl); 11
[0046] The compounds of this invention: (i) potently inhibit
farnesyl protein transferase, but not geranylgeranyl protein
transferase I, in vitro; (ii) block the phenotypic change induced
by a form of transforming Ras which is a farnesyl acceptor but not
by a form of transforming Ras engineered to be a geranylgeranyl
acceptor; (iii) block intracellular processing of Ras which is a
farnesyl acceptor but not of Ras engineered to be a geranylgeranyl
acceptor; and (iv) block abnormal cell growth in culture induced by
transforming Ras.
[0047] The compounds of this invention inhibit farnesyl protein
transferase and the farnesylation of the oncogene protein Ras.
Thus, this invention further provides a method of inhibiting
farnesyl protein transferase, (e.g., ras farnesyl protein
transferase) in mammals, especially humans, by the administration
of an effective amount of the tricyclic compounds described above.
The administration of the compounds of this invention to patients,
to inhibit farnesyl protein transferase, is useful in the treatment
of the cancers described below.
[0048] This invention provides a method for inhibiting or treating
the abnormal growth of cells. including transformed cells, by
administering an effective amount of a compound of this invention.
Abnormal growth of cells refers to cell growth independent of
normal regulatory mechanisms (e.g., loss of contact inhibition).
This includes the abnormal growth of: (1) tumor cells (tumors)
expressing an activated Ras oncogene; (2) tumor cells in which the
Ras protein is activated as a result of oncogenic mutation in
another gene; and (3) benign and malignant cells of other
proliferative diseases in which aberrant Ras activation occurs.
[0049] This invention also provides a method for inhibiting or
treating tumor growth by administering an effective amount of the
tricyclic compounds, described herein, to a mammal (e.g., a human)
in need of such treatment. In particular, this invention provides a
method for inhibiting or treating the growth of tumors expressing
an activated Ras oncogene by the administration of an effective
amount of the above described compounds. Examples of tumors which
may be inhibited or treated include, but are not limited to, lung
cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g.,
pancreatic carcinoma such as, for example, exocrine pancreatic
carcinoma), colon cancers (e.g., colorectal carcinomas, such as,
for example, colon adenocarcinoma and colon adenoma), myeloid
leukemias (for example, acute myelogenous leukemia (AML)), thyroid
follicular cancer, myelodysplastic syndrome (MDS), bladder
carcinoma, epidermal carcinoma, breast cancer and prostate
cancer.
[0050] It is believed that this invention also provides a method
for inhibiting or treating proliferative diseases, both benign and
malignant, wherein Ras proteins are aberrantly activated as a
result of oncogenic mutation in other genes--i.e., the Ras gene
itself is not activated by mutation to an oncogenic form--with said
inhibition or treatment being accomplished by the administration of
an effective amount of the tricyclic compounds described herein, to
a mammal (e.g., a human) in need of such treatment. For example,
the benign proliferative disorder neurofibromatosis, or tumors in
which Ras is activated due to mutation or overexpression of
tyrosine kinase oncogenes (e.g., neu, src, abl, lck, and fyn), may
be inhibited or treated by the tricyclic compounds described
herein.
[0051] The tricyclic compounds useful in the methods of this
invention inhibit or treat the abnormal growth of cells. Without
wishing to be bound by theory, it is believed that these compounds
may function through the inhibition of G-protein function, such as
ras p21, by blocking G-protein isoprenylation, thus making them
useful in the treatment of proliferative diseases such as tumor
growth and cancer. Without wishing to be bound by theory, it is
believed that these compounds inhibit ras farnesyl protein
transferase, and thus show antiproliferative activity against ras
transformed cells.
DETAILED DESCRIPTION OF THE INVENTION
[0052] As used herein, the following terms are used as defined
below unless otherwise indicated:
[0053] MH.sup.+--represents the molecular ion plus hydrogen of the
molecule in the mass spectrum:
[0054] Et (or ET)--represents ethyl (C.sub.2H.sub.5);
[0055] alkyl--represents straight and branched carbon chains and
contains from one to twenty carbon atoms, preferably one to six
carbon atoms;
[0056] halo-represents fluoro, chloro, bromo and iodo;
[0057] The following solvents and reagents are referred to herein
by the abbreviations indicated: ethanol (EtOH); methanol (MeOH);
acetic acid (HOAc or AcOH); ethyl acetate (EtOAc);
N,N-dimethylformamide (DMF); trifluoroacetic acid (TFA);
trifluoroacetic anhydride (TFAA); 1-hydroxybenzotriazole (HOBT);
1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (DEC);
diisobutylaluminum hydride(DIBAL); and 4-methylmorpholine
(NMM).
[0058] The positions in the tricyclic ring system are: 12
[0059] Preferred halo atoms for R.sup.1, R.sup.2, R.sup.3, and
R.sup.4 in Formula 1.0 are selected from: Br, Cl or I, with Br and
Cl being preferred.
[0060] Compounds of Formula 1.0 include compounds of the formula:
13
[0061] wherein R.sup.1 and R.sup.3 are the same or different halo.
Preferably, for these dihalo compounds, R.sup.1 and R.sup.3 are
independently selected from Br or Cl, and more preferably R.sup.1
is Br and R.sup.3 is Cl. Preferably, X is CH or N, with CH being
more preferred.
[0062] Compounds of Formula 1.0 include compounds of Formulas 1.1
and 1.2: 14
[0063] wherein R.sup.1, R.sup.3 and R.sup.4 in Formula 1.1 are
halo, and R.sup.1, R.sup.2 and R.sup.3 in Formula 1.2 are halo.
Compounds of Formula 1.1 are preferred.
[0064] Preferably, in Formula 1.1, R.sup.1 is Br, R.sup.3 is Cl,
and R.sup.4 is halo. More preferably, in Formula 1.1, R.sup.1 is
Br, R.sup.3 is Cl, and R.sup.4 is Br.
[0065] Preferably, in Formula 1.2, R.sup.1 is Br, R.sup.2 is halo,
and R.sup.3 is Cl. More preferably, in Formula 1.1, R.sup.1 is Br,
R.sup.2 is Br, and R.sup.3 is Cl.
[0066] Preferably, for compounds of Formulas 1.1 and 1.2, X is CH
or N. For compounds of Formula 1.1, X is preferably CH.
[0067] Preferably, for the compounds of this invention, the
optional bond between positions 5 and 6 (i.e., C5-C6) in the
tricyclic system is absent.
[0068] Also, preferably, for the compounds of this invention,
substituent a in Ring I represents N.
[0069] Those skilled in the art will appreciate that compounds of
Formula 1.0 include compounds of Formulas 1.3 and 1.4: 15
[0070] wherein X is CH or N, with compounds of 1.3 being preferred
for compounds of Formula 1.1, and with compounds of Formula 1.4
being preferred for componds of Formula 1.2.
[0071] Thus, compounds of the invention include compounds of the
formulas: 16
[0072] Compounds of Formula 1.9 are preferred.
[0073] Preferably substituent T is 17
[0074] More preferably, substituent T is the substituent of Formula
2.0 wherein the sum of b and d is 4. Most preferably b is 2 and d
is 2 forming the group: 18
[0075] Preferably, Y is O.
[0076] Examples of Formula 2.0 also include substituents wherein:
(a) the sum of b and d is 3, wherein b is 3 and d is 0; (b) the sum
of b and d is 4, wherein b is 4 and d is 0; (c) the sum of b and d
is 4, wherein b is 3 and d is 1; and (d) the sum of b and d is 3,
wherein b is 2 and d is 1. For these examples Y is preferably
O.
[0077] Examples of Formula 2.0 include: 19
[0078] includes substituents wherein: (a) the sum of e and f is 3,
wherein e is 3 and f is 0; (b) the sum of e and f is 2, wherein e
is l and d is 1; and (c) the sum of e and f is 2, wherein e is 2
and f is 0.
[0079] Examples of Formula 3.0 include: 20
[0080] includes substituents wherein: g is 0, h is 2, and i is 1.
Preferably, V and W are O. For example, Formula 4.0 includes the
substituent 21
[0081] includes the substituents: 22
[0082] includes the substituents: 23
[0083] Representative compounds of the invention include compounds
of the formula: 24
[0084] wherein R.sup.12 is selected from: 25
[0085] Those skilled in the art will appreciate that substituent
R.sup.12 is the same as substituent 26
[0086] Representative compounds of this invention also include:
27
[0087] Representative compounds of the invention also include:
28
[0088] Lines drawn into the ring systems indicate that the
indicated bond may be attached to any of the substitutable ring
carbon atoms.
[0089] Certain compounds of the invention may exist in different
isomeric (e.g., enantiomers and diastereoisomers) forms. The
invention contemplates all such isomers both in pure form and in
admixture, including racemic mixtures. Enol forms are also
included.
[0090] Certain tricyclic compounds will be acidic in nature, e.g.
those compounds which possess a carboxyl or phenolic hydroxyl
group. These compounds may form pharmaceutically acceptable salts.
Examples of such salts may include sodium, potassium, calcium,
aluminum, gold and silver salts. Also contemplated are salts formed
with pharmaceutically acceptable amines such as ammonia, alkyl
amines. hydroxyalkylamines, N-methylglucamine and the like.
[0091] Certain basic tricyclic compounds also form pharmaceutically
acceptable salts, e.g., acid addition salts. For example, the
pyrido-nitrogen atoms may form salts with strong acid, while
compounds having basic substituents such as amino groups also form
salts with weaker acids. Examples of suitable acids for salt
formation are hydrochloric, sulfuric, phosphoric, acetic, citric,
oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic,
maleic, methanesulfonic and other mineral and carboxylic acids well
known to those in the art. The salts are prepared by contacting the
free base form with a sufficient amount of the desired acid to
produce a salt in the conventional manner. The free base forms may
be regenerated by treating the salt with a suitable dilute aqueous
base solution such as dilute aqueous NaOH, potassium carbonate,
ammonia and sodium bicarbonate. The free base forms differ from
their respective salt forms somewhat in certain physical
properties, such as solubility in polar solvents, but the acid and
base salts are otherwise equivalent to their respective free base
forms for purposes of the invention.
[0092] All such acid and base salts are intended to be
pharmaceutically acceptable salts within the scope of the invention
and all acid and base salts are considered equivalent to the free
forms of the corresponding compounds for purposes of the
invention.
[0093] Compounds of the invention may be prepared according to the
procedures described in WO 95/10516 published Apr. 20, 1995, U.S.
Pat. No. 5,719,148 issued Feb. 17, 1998, and copending application
Ser. No. 08/766,601 filed Dec. 12, 1996; the disclosures of each
being incorporated herein by reference thereto; and according to
the procedures described below.
[0094] Compounds of the invention can be prepared according to the
reaction: 29
[0095] In the reaction, the cyclic ether carboxylic acid (14.0) is
coupled to the tricyclic amine (14.0) using amide bond forming
conditions well known to those skilled in the art. The substituents
are as defined for Formula 1.0. For example, carbodiimide coupling
methods (e.g., DEC) can be used. For example, the carboxylic acid
(14.0) can be reacted with the tricyclic amine (13.0) using
DEC/HOBT/NMM in DMF at about 25.degree. C. for a sufficient period
of time, e.g., about 18 hours, to produce a compound of Formula
1.0.
[0096] For example, using the carbodiimide coupling methods,
compounds of the invention can be produced according to the
reaction: 30
[0097] The cyclic ether carboxylic acids (14.0) are prepared by
methods well known in the art. Commercially available cyclic ether
ketones can be reacted in a Wittig reaction to produce
olefinesters. The olefin is then reduced by catalytic hydrogenation
or by metal hydride reduction to the saturated cyclic ether
acetates which are then hydrolyzed to the cyclic ether acids
(14.0). See, for example, J. Med. Chem. (1993), 36, 2300, the
disclosure of which is incorporated herein by reference thereto.
The reaction is illustrated in Scheme 1 below. 31
[0098] In Scheme 1, n represents 0 or 1, and Q represents O or
S.
[0099] The exocyclic olefin from the Wittig reaction in Scheme 1
can be reacted with cyanide in a Michael reaction to form a
nitrile, or with hydrogen peroxide to form an epoxide. The nitrile
can be hydrolyzed to a carboxy group and later converted to amides.
The epoxide can be hydrolyzed or reduced to an alcohol. This
reaction, well known to those skilled in the art, is illustrated in
Scheme 2 below. 32
[0100] wherein n and Q are as defined in Scheme 1.
[0101] The cyclic ether acetates can also be produced by the
insertion of an acetate carbene into a C--H bond next to the ether
heteroatom of a cyclic ether, as described in Tetrahedron (1989).
45, 69. The acetate carbene can be produced from a diazoacetate,
such as ethyl diazoacetate, and a rhodium or copper catalyst, such
as dirhodium diacetate of copper sulfate and heat. This is
illustrated by the reaction: 33
[0102] wherein n and Q are as defined in Scheme 1.
[0103] If the cyclic ether contains a double bond, the acetate
carbene can add to the double bond to produce a bicyclocyclic ether
acetate as described in Comp. Rend. (1957), 244, 2806. If the
double bond is adjacent to the ether heteroatom, the resulting
cyclopropyl ring can be opend by catalytic hydrogenation by an
alcohol and acid. This reaction is illustrated in Scheme 3 below.
34
[0104] wherein n and Q are as defined in Scheme 1.
[0105] Cyclic ethers containing a carboxy group directly attached
can be prepared by a base catalyzed cyclization of a dihalo ether
with diethyl malonate followed by hydrolysis and decarboxylation as
described in J. Am. Chem. Soc. (1995), 115, 8401. This is
illustrated by Scheme 4 below. 35
[0106] wherein n and Q are as defined in Scheme 1.
[0107] Many bicyclic-cyclic ether ketones are known in the
literature. Many of these can be made by Deils-Alder processes. For
example, J. Am. Chem. Soc. (1978), 100, 1765 describes the the
reaction: 36
[0108] These bicyclic-cyclic ether ketones can be reacted in a
Wittig reaction as above to produce bicyclic-cyclic ether
acetates.
[0109] Compounds of Formula 13.0a 37
[0110] are prepared by methods known in the art, for example by
methods disclosed in WO 95/10516, in U.S. Pat. No. 5,151,423 and
those described below. Compounds of Formula 13.0a wherein X is C
(when the double bond is present) or CH and the C-3 postion of the
pyridine ring in the tricyclic structure is substituted by bromo
(i.e., R.sup.1 is Br) can also be prepared by a procedure
comprising the following steps:
[0111] (a) reacting an amide of the formula 38
[0112] wherein R.sup.11a is Br, R.sup.5a is hydrogen and R.sup.6a
is C.sub.1-C.sub.6 alkyl, aryl or heteroaryl; R.sup.5a is
C.sub.1-C.sub.6 alkyl, aryl or heteroaryl and R.sup.6a is hydrogen;
R.sup.5a and R.sup.6a are independently selected from the group
consisting of C.sub.1-C.sub.6 alkyl and aryl; or R.sup.5a and
R.sup.6a, together with the nitrogen to which they are attached,
form a ring comprising 4 to 6 carbon atoms or comprising 3 to 5
carbon atoms and one hetero moiety selected from the group
consisting of --O-- and --NR.sup.9a--, wherein R.sup.9a is H,
C.sub.1-C.sub.6 alkyl or phenyl;
[0113] with a compound of the formula 39
[0114] wherein R.sup.1a, R.sup.2a, R.sup.3a and R.sup.4a are are
independently selected from the group consisting of hydrogen and
halo and R.sup.7a is Cl or Br, in the presence of a strong base to
obtain a compound of the formula 40
[0115] (b) reacting a compound of step (a) with
[0116] (i) POCl.sub.3 to obtain a cyano compound of the formula
41
[0117] (ii) DIBALH to obtain an aldehyde of the formula 42
[0118] (c) reacting the cyano compound or the aldehyde with a
piperidine derivative of the formula 43
[0119] wherein L is a leaving group selected from the group
consisting of Cl and Br, to obtain a ketone or an alcohol of the
formula below, respectively: 44
[0120] (d)(i) cyclizing the ketone with CF.sub.3SO.sub.3H to obtain
a compound of Formula 13.0a wherein the dotted line represents a
double bond; or
[0121] (d)(ii) cyclizing the alcohol with polyphosphoric acid to
obtain a compound of Formula 13.0a wherein the dotted line
represents a single bond.
[0122] Methods for preparing compounds of Formula 13.0a disclosed
in WO 95/10516, U.S. Pat. No. 5,151,423 and described below employ
a tricyclic ketone intermediate. Such intermediates of the formula
45
[0123] wherein R.sup.11b, R.sup.1a, R.sup.2a, R.sup.3a and R.sup.4a
are independently selected from the group consisting of hydrogen
and halo, can be prepared by the following process comprising:
[0124] (a) reacting a compound of the formula 46
[0125] (i) with an amine of the formula NHR.sup.5aR.sup.6a, wherein
R.sup.5a as R.sup.6a are as defined in the process above; in the
presence of a palladium catalyst and carbon monoxide to obtain an
amide of the formula: 47
[0126] (ii) with an alcohol of the formula R.sup.10aOH, wherein
R.sup.10a is C.sub.1-C.sub.6 lower alkyl or C.sub.3-C.sub.6
cycloalkyl, in the presence of a palladium catalyst and carbon
monoxide to obtain the ester of the formula 48
[0127] followed by reacting the ester with an amine of formula
NHR.sup.5aR.sup.6a to obtain the amide;
[0128] (b) reacting the amide with an iodo-substituted benzyl
compound of the formula 49
[0129] wherein R.sup.1a, R.sup.2a, R.sup.3a, R.sup.4a and R.sup.7a
are as defined above, in the presence of a strong base to obtain a
compound of the formula 50
[0130] (c) cyclizing a compound of step (b) with a reagent of the
formula R.sup.8aMgL, wherein R.sup.8a is C.sub.1-C.sub.8 alkyl,
aryl or heteroaryl and L is Br or Cl, provided that prior to
cyclization, compounds wherein R.sup.5a or R.sup.6a is hydrogen are
reacted with a suitable N-protecting group.
[0131] Compounds of Formula 1.0, wherein substituent a is NO (Ring
I) and X is C or CH, can be made from compounds of Formula 13.0a
using procedures well known to those skilled in the art. For
example the compound of Formula 13.0a can be reacted with
m-chloroperoxybenzoic acid in a suitable organic solvent, e.g.,
dichloromethane (usually anhydrous) or methylene chloride, at a
suitable temperature, to produce a compound of Formula 13.0b 51
[0132] Generally, the organic solvent solution of Formula 13.0a is
cooled to about 0.degree. C. before the m-chloroperoxybenzoic acid
is added. The reaction is then allowed to warm to room temperature
during the reaction period. The desired product can be recovered by
standard separation means. For example, the reaction mixture can be
washed with an aqueous solution of a suitable base, e.g., saturated
sodium bicarbonate or NaOH (e.g., 1N NaOH), and then dried over
anhydrous magnesium sulfate. The solution containing the product
can be concentrated in vacuo. The product can be purified by
standard means, e.g., by chromatography using silica gel (e.g.,
flash column chromatography).
[0133] Alternatively, compounds of Formula 1.0, wherein substituent
a is NO and X is C or CH, can be made from compounds of Formula
1.0, wherein substituent a is N, by the m-chloroperoxybenzoic acid
oxidation procedure described above.
[0134] Also, alternatively, the compounds of Formula 1.0, wherein
substituent a is NO and X is C or CH, can be made from tricyclic
ketone compounds 52
[0135] using the oxidation procedure with m-chloroperoxybenzoic
acid. The oxidized intermediate compounds 53
[0136] are then reacted by methods known in the art to produce
compounds of the invention.
[0137] Those skilled in the art will appreciate that the oxidation
reaction can be conducted on racemic mixtures and the isomers can
then be separated by know techniques, or the isomers can be
separated first and then oxidized to the corresponding N-oxide.
[0138] Those skilled in the art will appreciate that it is
preferable to avoid an excess of m-chloroperoxybenzoic acid when
the oxidation reaction is carried out on the compounds having a
C-11 double bond to piperidine Ring IV. In these reactions an
excess of m-chloroperoxybenzoic acid can cause epoxidation of the
C-11 double bond.
[0139] (+)-Isomers of compounds of Formula 13.0a wherein X is CH
can be prepared with high enantioselectivity by using a process
comprising enzyme catalyzed transesterification. Preferably, a
racemic compound of Formula 13.0a, wherein X is C, the double bond
is present and R.sup.4 is not H, is reacted with an enzyme such as
Toyobo LIP-300 and an acylating agent such as trifluoroethly
isobutyrate; the resultant (+)-amide is then hydrolyzed, for
example by refluxing with an acid such as H.sub.2SO.sub.4, to
obtain the corresponding optically enriched (+)-isomer wherein X is
CH and R.sup.3 is not H. Alternatively, a racemic compound of
Formula 13.0a, wherein X is C, the double bond is present and
R.sup.4 is not H, is first reduced to the corresponding racemic
compound of Formula 13.0a wherein X is CH and then treated with the
enzyme (Toyobo LIP-300) and acylating agent as described above to
obtain the (+)-amide, which is hydrolyzed to obtain the optically
enriched (+)-isomer.
[0140] Compounds of the invention, wherein a is NO and X is N, can
be prepared from the tricyclic ketone (II) described above. Ketone
(II) can be converted to the corresponding C-11 hydroxy compound
which in turn can be converted to the corresponding C-11 chloro
compound 54
[0141] and (IV) can then be reacted with piperazine to produce the
intermediate 55
[0142] Intermediate (V) can then be reacted with the reagents,
using techniques well known in the art, which will provide the
desired compound.
[0143] Compounds useful in this invention are exemplified by the
following examples, which should not be construed to limit the
scope of the disclosure.
PREPARATIVE EXAMPLE 1
[0144] 56 57
[0145] Combine 14.95 g (39 mmol) of
8-chloro-11-(1-ethoxy-carbonyl-4-piper-
idinyl)-11H-benzo[5,6]cyclohepta[1,2-b]pyridine and 150 mL of
CH.sub.2Cl.sub.2, then add 13.07 g (42.9 mmol) of
(nBu).sub.4NNO.sub.3 and cool the mixture to 0.degree. C. Slowly
add (dropwise) a solution of 6.09 mL (42.9 mmol) of TFAA in 20 mL
of CH.sub.2Cl.sub.2 over 1.5 hours. Keep the mixture at 0.degree.
C. overnight, then wash successively with saturated NaHCO.sub.3
(aqueous), water and brine. Dry the organic solution over
Na.sub.2SO.sub.4, concentrate in vacuo to a residue and
chromatograph the residue (silica gel, EtOAc/hexane gradient) to
give 4.32 g and 1.90 g of the two product compounds 1A(i) and
1A(ii), respectively. Mass Spec. for compound 1A(i):
MH.sup.+=428.2. Mass Spec. for compound 1A(ii): MH.sup.+=428.3.
58
[0146] Combine 22.0 g (51.4 mmol) of the product 1A(i) from Step A,
150 mL of 85% EtOH (aqueous), 25.85 g (0.463 mole) of Fe powder and
2.42 g (21.8 mmol) of CaCl.sub.2, and heat at reflux overnight. Add
12.4 g (0.222 mole) of Fe powder and 1.2 g (10.8 mmol) of
CaCl.sub.2 and heat at reflux for 2 hours. Add another 12.4 g
(0.222 mole) of Fe powder and 1.2 g (10.8 mmol) of CaCl.sub.2 and
heat at reflux for 2 hours more. Filter the hot mixture through
celite.RTM., wash the celite.RTM. with 50 mL of hot EtOH and
concentrate the filtrate in vacuo to a residue. Add 100 mL of
anhydrous EtOH, concentrate to a residue and chromatograph the
residue (silica gel, MeOH/CH.sub.2Cl.sub.2 gradient) to give 16.47
g of the product compound. 59
[0147] Combine 16.47 g (41.4 mmol) of the product from Step B. and
150 mL of 48% HBr (aqueous) and cool to -3.degree. C. Slowly add
(dropwise) 18 mL of bromine, then slowly add (dropwise) a solution
of 8.55 g (0.124 mole) of NaNO.sub.2 in 85 mL of water. Stir for 45
minutes at -3.degree. to 0.degree. C., then adjust to pH=10 by
adding 50% NaOH (aqueous). Extract with EtOAc, wash the extracts
with brine and dry the extracts over Na.sub.2SO.sub.4. Concentrate
to a residue and chromatograph (silica gel, EtOAc/hexane gradient)
to give 10.6 g and 3.28 g of the two product compounds 1C(i) and
1C(ii), respectively. Mass Spec. for compound 1C(i): MH.sup.+=
461.2. Mass Spec. for compound 1C(ii): MH.sup.+=539. 60
[0148] Hydrolyze the product 3C(i) of Step C by dissolving in
concentrated HCl and heating to about 100.degree. C. for @ 16
hours. Cool the mixture, the neutralize with 1 M NaOH (aqueous).
Extract with CH.sub.2Cl.sub.2, dry the extracts over MgSO.sub.4,
filter and concentrate in vacuo to the title compound. Mass Spec.:
MH.sup.+= 466.9.
PREPARATIVE EXAMPLE 2
[0149] 61
[0150] Combine 25.86 g (55.9 mmol) of
4-(8-chloro-3-bromo-5,6-dihydro-11H--
benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic
acid ethyl ester and 250 mL of concentrated H.sub.2SO.sub.4 at
-5.degree. C., then add 4.8 g (56.4 mmol) of NaNO.sub.3 and stir
for 2 hours. Pour the mixture into 600 g of ice and basify with
concentrated NH.sub.4OH (aqueous). Filter the mixture, wash with
300 mL of water, then extract with 500 mL of CH.sub.2Cl.sub.2. Wash
the extract with 200 mL of water, dry over MgSO.sub.4, then filter
and concentrate in vacuo to a residue. Chromatograph the residue
(silica gel, 10% EtOAc/CH.sub.2Cl.sub.2) to give 24.4 g (86% yield)
of the product. m.p.=165-167.degree. C., Mass Spec.: MH.sup.+=506
(CI). Elemental analysis: calculated--C, 52.13; H, 4.17; N, 8.29;
found--C, 52.18; H, 4.51; N, 8.16. 62
[0151] Combine 20 g (40.5 mmol) of the product of Step A and 200 mL
of concentrated H.sub.2SO.sub.4 at 20.degree. C., then cool the
mixture to 0.degree. C. Add 7.12 g (24.89 mmol) of
1,3-dibromo-5,5-dimethylhydantoin to the mixture and stir for 3
hours at 20.degree. C. Cool to 0.degree. C., add an additional 1.0
g (3.5 mmol) of the dibromohydantoin and stir at 20.degree. C. for
2 hours. Pour the mixture into 400 g of ice, basify with
concentrated NH.sub.4OH (aqueous) at 0.degree. C., and collect the
resulting solid by filtration. Wash the solid with 300 mL of water,
slurry in 200 mL of acetone and filter to provide 19.79 g (85.6%
yield) of the product. m.p.=236-237.degree. C., Mass Spec.:
MH.sup.+=584 (CI). Elemental analysis: calculated--C, 45.11; H.
3.44; N, 7.17; found--C, 44.95; H, 3.57; N, 7.16
[0152] Step C: 63
[0153] Combine 25 g (447 mmol) of Fe filings, 10 g (90 mmol) of
CaCl.sub.2 and a suspension of 20 g (34.19 mmol) of the product of
Step B in 700 mL of 90:10 EtOH/water at 50.degree. C. Heat the
mixture at reflux overnight, filter through Celite.RTM. and wash
the filter cake with 2.times.200 mL of hot EtOH. Combine the
filtrate and washes, and concentrate in vacuo to a residue. Extract
the residue with 600 mL of CH.sub.2Cl.sub.2, wash with 300 mL of
water and dry over MgSO.sub.4. Filter and concentrate in vacuo to a
residue, then chromatograph (silica gel, 30%
EtOAc/CH.sub.2Cl.sub.2) to give 11.4 g (60% yield) of the product.
m.p.=211-212.degree. C., Mass Spec.: MH.sup.+= 554 (CI). Elemental
analysis: calculated--C, 47.55; H, 3.99; N, 7.56; found--C, 47.45;
H, 4.31; N. 7.49. 64
[0154] Slowly add (in portions) 20 g (35.9 mmol) of the product of
Step C to a solution of 8 g (116 mmol) of NaNO.sub.2 in 120 mL of
concentrated HCl (aqueous) at -10.degree. C. Stir the resulting
mixture at 0.degree. C. for 2 hours, then slowly add (dropwise) 150
mL (1.44 mole) of 50% H.sub.3PO.sub.2 at 0.degree. C. over a 1 hour
period. Stir at 0.degree. C. for 3 hours, then pour into 600 g of
ice and basify with concentrated NH.sub.4OH (aqueous). Extract with
2.times.300 mL of CH.sub.2Cl.sub.2, dry the extracts over
MgSO.sub.4, then filter and concentrate in vacuo to a residue.
Chromatograph the residue (silica gel, 25% EtOAc/hexanes) to give
13.67 g (70% yield) of the product. m.p.=163-165.degree. C., Mass
Spec.: MH.sup.+=539 (CI). Elemental analysis: calculated--C, 48.97;
H, 4.05; N, 5.22; found--C, 48.86; H, 3.91; N, 5.18. 65
[0155] Combine 6.8 g (12.59 mmol) of the product of Step D and 100
mL of concentrated HCl (aqueous) and stir at 85.degree. C.
overnight. Cool the mixture, pour it into 300 g of ice and basify
with concentrated NH.sub.4OH (aqueous). Extract with 2.times.300 mL
of CH.sub.2Cl.sub.2, then dry the extracts over MgSO.sub.4. Filter,
concentrate in vacuo to a residue, then chromatograph (silica gel,
10% MeOH/EtOAc+2% NH.sub.4OH (aqueous)) to give 5.4 g (92% yield)
of the title compound. m.p.=172-174.degree. C., Mass Spec.: MH+=467
(FAB). Elemental analysis: calculated--C, 48.69; H, 3.65; N, 5.97;
found--C, 48.83; H, 3.80; N, 5.97
PREPARATIVE EXAMPLE 3
[0156] 66 67
[0157] Hydrolyze 2.42 g of
4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]c-
yclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic acid
ethyl ester via substantially the same procedure as described in
Preparative Example 1, Step D, to give 1.39 g (69% yield) of the
product. 68
[0158] Combine 1 g (2.48 mmol) of the product of Step A and 25 mL
of dry toluene, add 2.5 mL of 1 M DIBAL in toluene and heat the
mixture at reflux. After 0.5 hours, add another 2.5 mL of 1 M DIBAL
in toluene and heat at reflux for 1 hour. (The reaction is
monitored by TLC using 50% MeOH/CH.sub.2Cl.sub.2+NH.sub.4OH
(aqueous).) Cool the mixture to room temperature, add 50 mL of 1 N
HCl (aqueous) and stir for 5 min. Add 100 mL of 1 N NaOH (aqueous),
then extract with EtOAc (3.times.150 mL). Dry the extracts over
MgSO.sub.4, filter and concentrate in vacuo to give 1.1 g of the
title compound.
PREPARATIVE EXAMPLE 4
[0159] 69
[racemic as well as (+)- and (-)-isomers]
[0160] 70
[0161] Combine 16.6 g (0.03 mole) of the product of Preparative
Example 2, Step D, with a 3:1 solution of CH.sub.3CN and water
(212.65 mL CH.sub.3CN and 70.8 mL of water) and stir the resulting
slurry overnight at room temperature. Add 32.833 g (0.153 mole) of
NaIO.sub.4 and then 0.31 g (2.30 mmol) of RuO2 and stir at room
temperature give 1.39 g (69% yield) of the product. (The addition
of RuO is accompanied by an exothermnic reaction and the
temperature climbs from 20.degree. to 30.degree. C.) Stir the
mixture for 1.3 hrs. (temperature returned to 25.degree. C. after
about 30 min.), then filter to remove the solids and wash the
solids with CH.sub.2Cl.sub.2. Concentrate the filtrate in vacuo to
a residue and dissolve the residue in CH.sub.2Cl.sub.2. Filter to
remove insoluble solids and wash the solids with CH.sub.2Cl.sub.2.
Wash the filtrate with water, concentrate to a volume of about 200
mL and wash with bleach, then with water. Extract with 6 N HCl
(aqueous). Cool the aqueous extract to 0.degree. C. and slowly add
50% NaOH (aqueous) to adjust to pH=4 while keeping the temperature
<30.degree. C. Extract twice with CH.sub.2Cl.sub.2, dry over
MgSO.sub.4 and concentrate in vacuo to a residue. Slurry the
residue in 20 mL of EtOH and cool to 0.degree. C. Collect the
resulting solids by filtration and dry the solids in vacuo to give
7.95 g of the product. .sup.1H NMR (CDCl.sub.3, 200 MHz): 8.7 (s,
1H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45 (m, 2H); 3.15 (m, 2H). 71
[0162] Combine 21.58 g (53.75 mmol) of the product of Step A and
500 mL of an anhydrous 1:1 mixture of EtOH and toluene, add 1.43 g
(37.8 mmol) of NaBH.sub.4 and heat the mixture at reflux for 10
min. Cool the mixture to 0.degree. C., add 100 mL of water, then
adjust to pH.apprxeq.4-5 with 1 M HCl (aqueous) while keeping the
temperature <10.degree. C. Add 250 mL of EtOAc and separate the
layers. Wash the organic layer with brine (3.times.50 mL) then dry
over Na.sub.2SO.sub.4. Concentrate in vacuo to a residue (24.01 g)
and chromatograph the residue (silica gel, 30%
hexane/CH.sub.2Cl.sub.2) to give the product. Impure fractions were
purified by rechromatography. A total of 18.57 g of the product was
obtained. .sup.1H NMR (DMSO-d.sub.6, 400 MHz): 8.5 (s, 1H); 7.9 (s,
1H); 7.5 (d of d, 2H); 6.2 (s, 1H); 6.1 (s, 1H); 3.5 (m, 1H); 3.4
(m, 1H); 3.2 (m, 2H). 72
[0163] Combine 18.57 g (46.02 mmol) of the product of Step B and
500 mL of CHCl.sub.3, then add 6.70 mL (91.2 mmol) of SOCl.sub.2,
and stir the mixture at room temperature for 4 hrs. Add a solution
of 35.6 g (0.413 mole) of piperazine in 800 mL of THF over a period
of 5 min. and stir the mixture for 1 hr. at room temperature. Heat
the mixture at reflux overnight, then cool to room temperature and
dilute the mixture with 1 L of CH.sub.2Cl.sub.2. Wash with water
(5.times.200 mL), and extract the aqueous wash with CHCl.sub.3
(3.times.100 mL). Combine all of the organic solutions, wash with
brine (3.times.200 mL) and dry over MgSO.sub.4. Concentrate in
vacuo to a residue and chromatograph (silica gel, gradient of 5%,
7.5%, 10% MeOH/CH.sub.2Cl.sub.2+NH.sub.4OH) to give 18.49 g of the
title compound as a racemic mixture. 73
[0164] The racemic title compound of Step C is separated by
preparative chiral chromatography (Chiralpack AD, 5 cm.times.50 cm
column, flow rate 100 mL/min., 20% iPrOH/hexane+0.2% diethylamine),
to give 9.14 g of the (+)-isomer and 9.30 g of the (-)-isomer.
[0165] Physical chemical data for (+)-isomer:
m.p.=74.5.degree.-77.5.degre- e. C.; Mass Spec. MH.sup.+=471.9;
[.alpha.].sub.D.sup.25=+97.4.degree. (8.48 mg/2 mL MeOH).
[0166] Physical chemical data for (-)-isomer:
m.p.=82.9.degree.-84.5.degre- e. C.; Mass Spec. MH.sup.+=471.8;
[.alpha.].sub.D.sup.25=+97.4.degree. (8.32 mg/2mL MeOH).
PREPARATIVE EXAMPLE 5
[0167] 74
[0168] Combine 15 g (38.5 mmol) of
4-(8-chloro-3-bromo-5,6-dihydro-11H-ben-
zo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic
acid ethyl ester and 150 mL of concentrated H.sub.2SO.sub.4 at
-5.degree. C., then add 3.89 g (38.5 mmol) of KNO.sub.3 and stir
for 4 hours. Pour the mixture into 3 L of ice and basify with 50%
NaOH (aqueous). Extract with CH.sub.2Cl.sub.2, dry over MgSO.sub.4,
then filter and concentrate in vacuo to a residue. Recrystallize
the residue from acetone to give 6.69 g of the product. .sup.1H NMR
(CDCl.sub.3, 200 MHz): 8.5 (s, 1H); 7.75 (s, 1H): 7.6 (s, 1H); 7.35
(s, 1H); 4.15 (q, 2H); 3.8 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.8 (m,
2H); 2.6-2.2 (m, 4H); 1.25 (t, 3H). 75
[0169] Combine 6.69 g (13.1 mmol) of the product of Step A and 100
mL of 85% EtOH/water, then add 0.66 g (5.9 mmol) of CaCl.sub.2 and
6.56 g (117.9 mmol) of Fe and heat the mixture at reflux overnight.
Filter the hot reaction mixture through celite.RTM. and rinse the
filter cake with hot EtOH. Concentrate the filtrate in vacuo to
give 7.72 g of the product. Mass Spec.: MH.sup.+=478.0 76
[0170] Combine 7.70 g of the product of Step B and 35 mL of HOAc,
then add 45 mL of a solution of Br2 in HOAc and stir the mixture at
room temperature overnight. Add 300 mL of 1 N NaOH (aqueous), then
75 mL of 50% NaOH (aqueous) and extract with EtOAc. Dry the extract
over MgSO.sub.4 and concentrate in vacuo to a residue.
Chromatograph the residue (silica gel, 20%-30% EtOAc/hexane) to
give 3.47 g of the product (along with another 1.28 g of partially
purified product). Mass Spec.: MH.sup.+= 555.9.
[0171] .sup.1H NMR (CDCl.sub.3, 300 MHz): 8.5 (s, 1H); 7.5 (s, 1H);
7.15 (s, 1H); 4.5 (s, 2H); 4.15 (m, 3H); 3.8 (br s, 2H); 3.4-3.1
(m, 4H); 9-2.75 (m, 1H); 2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H); 1.25 (m,
3H). 77
[0172] Combine 0.557 g (5.4 mmol) of t-butylnitrite and 3 mL of
DMF, and heat the mixture at to 60.degree.-70.degree. C. Slowly add
(dropwise) a mixture of 2.00 g (3.6 mmol) of the product of Step C
and 4 mL of DMF, then cool the mixture to room temperature. Add
another 0.64 mL of t-butylnitrite at 40.degree. C. and reheat the
mixture to 60.degree.-70.degree. C. for 0.5 hrs. Cool to room
temperature and pour the mixture into 150 mL of water. Extract with
CH.sub.2Cl.sub.2, dry the extract over MgSO.sub.4 and concentrate
in vacuo to a residue. Chromatograph the residue (silica gel,
10%-20% EtOAc/hexane) to give 0.74 g of the product. Mass Spec.:
MH.sup.+=541.0.
[0173] .sup.1H NMR (CDCl.sub.3, 200 MHz): 8.52 (s, 1H); 7.5 (d,
2H); 7.2 (s, 1H); 4.15 (q, 2H); 3.9-3.7 (m, 2H); 3.5-3.1 (m, 4H);
3.0-2.5 (m, 2H); 2.4-2.2 (m, 2H); 2.1-1.9 (m, 2H); 1.26 (t, 3H).
78
[0174] Combine 0.70 g (1.4 mmol) of the product of Step D and 8 mL
of concentrated HCl (aqueous) and heat the mixture at reflux
overnight. Add 30 mL of 1 N NaOH (aqueous), then 5 mL of 50% NaOH
(aqueous) and extract with CH.sub.2Cl.sub.2. Dry the extract over
MgSO.sub.4 and concentrate in vacuo to give 0.59 g of the title
compound. Mass Spec.: M.sup.+=468.7.
m.p.=123.9.degree.-124.2.degree. C.
PREPARATIVE EXAMPLE 6
[0175] 79
[racemic as well as (+)- and (-)-isomers]
[0176] 80
[0177] Prepare a solution of 8.1 g of the title compound from
Preparative Example 5. Step E, in toluene and add 17.3 mL of a 1M
solution of DIBAL in toluene. Heat the mixture at reflux and slowly
add (dropwise) another 21 mL of 1 M DIBAL/toluene solution over a
period of 40 min. Cool the reaction mixture to about 0.degree. C.
and add 700 mL of 1 M HCl (aqueous). Separate and discard the
organic phase. Wash the aqueous phase with CH.sub.2Cl.sub.2,
discard the extract, then basify the aqueous phase by adding 50%
NaOH (aqueous). Extract with CH.sub.2Cl.sub.2, dry the extract over
MgSO.sub.4 and concentrate in vacuo to give 7.30 g of the title
compound, which is a racemic mixture of enantiomers. 81
[0178] The racemic title compound of Step A is separated by
preparative chiral chromatography (Chiralpack AD, 5 cm.times.50 cm
column, using 20% iPrOH/hexane+0.2% diethylamine), to give the
(+)-isomer and the (-)-isomer of the title compound.
[0179] Physical chemical data for (+)-isomer: m.p.=148.8.degree.
C.; Mass Spec. MH.sup.+=469; [.alpha.].sub.D.sup.25=+65.6.degree.
(12.93 mg/2mL MeOH).
[0180] Physical chemical data for (-)-isomer: m.p.=112.degree. C.;
Mass Spec. MH.sup.+=469; [.alpha.].sub.D.sup.25=-65.2.degree. (3.65
mg/2mL MeOH).
PREPARATIVE EXAMPLE 7
[0181] 82
[racemic as well as (+)- and (-)-isomers]
[0182] 83
[0183] Combine 40.0 g (0.124 mole) of the starting ketone and 200
mL of H.sub.2SO.sub.4 and cool to 0.degree. C. Slowly add 13.78 g
(0.136 mole) of KNO.sub.3 over a period of 1.5 hrs., then warm to
room temperature and stir overnight. Work up the reaction using
substantially the same procedure as described for Preparative
Example 2, Step A. Chromatograph (silica gel, 20%, 30%, 40%. 50%
EtOAc/hexane, then 100% EtOAc) to give 28 g of the 9-nitro product,
along with a smaller quantity of the 7-nitro product and 19 g of a
mixture of the 7-nitro and 9-nitro compounds. 84
[0184] React 28 g (76.2 mmol) of the 9-nitro product of Step A, 400
mL of 85% EtOH/water, 3.8 g (34.3 mmol) of CaCl.sub.2 and 38.28 g
(0.685 mole) of Fe using substantially the same procedure as
described for Preparative Example 2, Step C, to give 24 g of the
product 85
[0185] Combine 13 g (38.5 mmol) of the product of Step B, 140 mL of
HOAc and slowly add a solution of 2.95 mL (57.8 mmol) of Br.sub.2
in 10 mL of HOAc over a period of 20 min. Stir the reaction mixture
at room temperature, then concentrate in vacuo to a residue. Add
CH.sub.2Cl.sub.2 and water, then adjust to pH=8-9 with 50% NaOH
(aqueous). Wash the organic phase with water, then brine and dry
over Na.sub.2SO.sub.4. Concentrate in vacuo to give 11.3 g of the
product. 86
[0186] Cool 100 mL of concentrated HCl (aqueous) to 0.degree. C.,
then add 5.61 g (81.4 mmol) of NaNO.sub.2 and stir for 10 min.
Slowly add (in portions) 11.3 g (27.1 mmol) of the product of Step
C and stir the mixture at 0.degree.-3.degree. C. for 2.25 hrs.
Slowly add (dropwise) 180 mL of 50% H.sub.3PO.sub.2 (aqueous) and
allow the mixture to stand at 0.degree. C. overnight. Slowly add
(dropwise) 150 mL of 50% NaOH over 30 min., to adjust to pH=9, then
extract with CH.sub.2Cl.sub.2. Wash the extract with water, then
brine and dry over Na.sub.2SO.sub.4. Concentrate in vacuo to a
residue and chromatograph (silica gel, 2% EtOAc/CH.sub. 2Cl.sub.2)
to give 8.6 g of the product. 87
[0187] Combine 8.6 g (21.4 mmol) of the product of Step D and 300
mL of MeOH and cool to 0.degree.-2.degree. C. Add 1.21 g (32.1
mmol) of NaBH.sub.4 and stir the mixture at .about.0.degree. C. for
1 hr. Add another 0.121 g (3.21 mmol) of NaBH.sub.4, stir for 2 hr.
at 0.degree. C., then let stand overnight at 0.degree. C.
Concentrate in vacuo to a residue then partition the residue
between CH.sub.2Cl.sub.2 and water. Separate the organic phase and
concentrate in vacuo (50.degree. C.) to give 8.2 g of the product.
88
[0188] Combine 8.2 g (20.3 mmol) of the product of Step E and 160
mL of CH.sub.2Cl.sub.2, cool to 0.degree. C., then slowly add
(dropwise) 14.8 mL (203 mmol) of SOCl.sub.2 over a 30 min. period.
Warm the mixture to room temperature and stir for 4.5 hrs., then
concentrate in vacuo to a residue, add CH.sub.2Cl.sub.2 and wash
with 1 N NaOH (aqueous) then brine and dry over Na2SO.sub.4.
Concentrate in vacuo to a residue, then add dry THF and 8.7 g (101
mmol) of piperazine and stir at room temperature overnight.
Concentrate in vacuo to a residue, add CH.sub.2Cl.sub.2, and wash
with 0.25 N NaOH (aqueous), water, then brine. Dry over
Na.sub.2SO.sub.4 and concentrate in vacuo to give 9.46 g of the
crude product. Chromatograph (silica gel, 5%
MeOH/CH.sub.2Cl.sub.2+NH.sub.3) to give 3.59 g of the title
compound, as a racemate. .sup.1H NMR (CDCl.sub.3, 200 MHz): 8.43
(d, 1H); 7.55 (d, 1H); 7.45 (d, 1H); 7.11 (d, 1H); 5.31 (s, 1H);
4.86-4.65 (m, 1H); 3.57-3.40 (m, 1H); 2.98-2.55 (m, 6H); 2.45-2.20
(m, 5H). 89
[0189] The racemic title compound from Step F (5.7 g) is
chromatographed as described for Preparative Example 4, Step D,
using 30% iPrOH/hexane+0.2% diethylamine, to give 2.88 g of the
R-(+)-isomer and 2.77 g of the S-(-)-isomer of the title
compound.
[0190] Physical chemical data for the R-(+)-isomer: Mass Spec.
MH.sup.+=470.0; [.alpha.].sub.D.sup.25=+12.1.degree. (10.9 mg/2 mL
MeOH).
[0191] Physical chemical data for the S-(-)-isomer: Mass Spec.
MH.sup.+=470.0; [.alpha.].sub.D.sup.25=+13.2.degree. (11.51 mg/2 mL
MeOH).
PREPARATIVE EXAMPLE 8
[0192] 90
[racemic as well as (+)- and (-)-isomers]
[0193] 91
[0194] Combine 13 g (33.3 mmol) of the title compound from
Preparative Example 2, Step E, and 300 mL of toluene at 20.degree.
C., then add 32.5 mL (32.5 mmol) of a 1 M solution of DIBAL in
toluene. Heat the mixture at reflux for 1 hr., cool to 20.degree.
C., add another 32.5 mL of 1 M DIBAL solution and heat at reflux
for 1 hr. Cool the mixture to 20.degree. C. and pour it into a
mixture of 400 g of ice, 500 mL of EtOAc and 300 mL of 10% NaOH
(aqueous). Extract the aqueous layer with CH.sub.2Cl.sub.2
(3.times.200 mL), dry the organic layers over MgSO.sub.4, then
concentrate in vacuo to a residue. Chromatograph (silica gel, 12%
MeOH/CH.sub.2Cl.sub.2+4% NH.sub.4OH) to give 10.4 g of the title
compound as a racemate. Mass Spec.: MH.sup.+=469 (FAB). Partial
.sup.1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H); 7.57 (s, 1H): 7.27
(d. 1H): 7.06 (d, 1H), 3.95 (d, 1H). 92
[0195] The racemic title compound of Step A is separated by
preparative chiral chromatography (Chiralpack AD, 5 cm.times.50 cm
column, using 5% iPrOH/hexane+0.2% diethylamine), to give the
(+)-isomer and the (-)-isomer of the title compound.
[0196] Physical chemical data for (+)-isomer: Mass Spec.
MH.sup.+=469 (FAB); [.alpha.].sub.D.sup.25=+43.5.degree. (c=0.402,
EtOH); partial .sup.1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H);
7.57 (s, 1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1H).
[0197] Physical chemical data for (-)-isomer: Mass Spec.
MH.sup.+=469 (FAB); [.alpha.].sub.D.sup.25=-41.8.degree. (c=0.328
EtOH); partial .sup.1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H);
7.57 (s, 1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1H).
PREPARATIVE EXAMPLE 9
[0198] 93
[racemic as well as R-(+)- and S-(-)-isomers]
[0199] The compound 94
[0200] is prepared according to the procedures of Preparative
Example 40 of WO 95/10516 (published Apr. 20, 1995), by following
the procedures described in Example 193 of WO 95/10516.
[0201] The (+)- and (-)-isomers can be separated by following
essentially the same procedure as Step D of Preparative Example
4.
[0202] Physical chemical data for the R-(+)-isomer: .sup.13C NMR
(CDCl.sub.3): 155.8 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2
(C); 135.3 (C); 133.4 (C); 132.0 (CH); 129.9 (CH); 125.6 (CH);
119.3 (C); 79.1 (CH); 52.3 (CH.sub.2); 52.3 (CH); 45.6 (CH.sub.2);
45.6 (CH.sub.2); 30.0 (CH.sub.2); 29.8 (CH.sub.2).
[.alpha.].sub.D.sup.25=+25.8.degree. (8.46 mg/2 mL MeOH).
[0203] Physical chemical data for the S-(-)-isomer: .sup.13C NMR
(CDCl.sub.3): 155.9 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2
(C); 135.3 (C); 133.3 (C); 132.0 (CH); 129.9 (CH); 125.5 (CH);
119.2 (C); 79.1 (CH); 52.5 (CH.sub.2); 52.5 (CH); 45.7 (CH.sub.2);
45.7 (CH.sub.2); 30.0 (CH.sub.2); 29.8 (CH.sub.2).
[.alpha.].sub.D.sup.25=-27.9.degree. (8.90 mg/2 mL MeOH).
PREPARATIVE EXAMPLE 10
Ethyl tetrahydropyran-4-ylidenylacetate (15.0), and ethyl
5,6-dihydro-2H-pyran-4-acetate (16.0)
[0204] 95
[0205] Following the chemistry described in J. Med. Chem., (1993),
36, 2300, a 2 L three-neck flask equipped with a thermometer,
addition funnel and a nitrogen inlet tube and a magnetic stirrer
was flame dried and charged with 1.0 L of anhydrous
1,2-dimethoxyethane and 9.0 g (0.38 mol) of sodium hydride (60%
dispersion in oil). Triethyl phosphono-acetate, 56 g (0.25 mol),
was added, dropwise with sirring, at such a rate that the reaction
temperature was maintained at 20-25.degree. C. After addition, the
reaction was stirred at 25.degree. C. for 45 min. then 25 g (0.25
mol) of tetrahydro-4H-pyran-4-one was added dropwise while keeping
the reaction temperature at 20-25.degree. C. by cooling with an ice
bath. After addition, the reaction was refluxed for one hour,
cooled to room temperature and then poured into 4 L of ice water.
This was extracted with three 2 L portions of ether. The combined
ether layers were dried over magnesium sulfate and concentrated
under vacuum giving 27 g of a yellow oil that is a 1:1.4 mixture of
15.0 and 16.0 as determined by NMR.
[0206] Sixteen grams of the above oil were flash chromatographed on
1.5 Kg of silica gel using ethyl acetate-hexane, 10-90, and
collecting 200 mL fractions. Fractions 13-22 yielded 5.65 g of pure
15.0, ethyl tetrahydropyran-4-ylidenyl-acetate, and fractions 31-50
yielded 8.06 g of pure 16.0, ethyl
5,6-dihydro-2H-pyran-4-acetate.
PREPARATIVE EXAMPLE 11
Ethyl tetrahydropyran-4-acetate
[0207] 96
[0208] A mixture of 15.0 and 16.0 (3 g, 17.6 mmol) from Preparative
Example 10 was dissolved in 20 mL of ethyl acetate containing 1.0 g
of 10% paladium on carbon. This mixture was stirred for 18 hours
under an atmosphere of hydrogen. The catalyst was filtered and the
filtrate was concentrated under vacuum giving 3.04 g of the title
product as a colorless oil.
PREPARATIVE EXAMPLE 12
Ethyl tetrahydrothiopyran-4-ylidenylacetate
[0209] 97
[0210] Following the procedure of Preparative Example 10, but using
2.32 g (20 mmol) of tetrahydrothiopyran-4-one instead of
tetrahydropyran-4-one, 3.53 g of the product was obtained as a
colorless oil.
PREPARATIVE EXAMPLE 13
Ethyl tetrahydrothiopyran-4-acetate
[0211] 98
[0212] Ethyl tetrahydrothiopyran-4-ylidenylacetate (2.3 g, 12.4
mmol), from Preparative Example 12, was dissolved in 25 mL of
ethanol containing 2.34 g (61.8 mmol) of sodium borohydride. After
stirring for 24 hours at 25.degree. C., an additional 1.2 g of
sodium borohydride was added and the reaction was stirred for an
additional 24 hours. Two additional 1.2 g portions of sodium
borohydride were added followed by stirring for 24 hours after each
addition. Silica gel TLC using hexane-ethyl acetate (95-5) showed
the reaction to be complete. The reaction was treated with 200 mL
of water and stirred for 5 minutes. The mixture was then extracted
with three 150 mL portions of ethyl acetate. The combined organic
layers were dried over magnesium sulfate and concentrated under
vacuum giving 1.6 g of a colorless oil. The oil was chromatographed
on 325 mL of silica gel using hexane-ethyl acetate (98-2) and 125
mL fractions were collected. Fractions 2-15 yielded 0.24 g of the
product as a colorless oil.
PREPARATIVE EXAMPLE 14
Ethyl 2'-(1,4-dioxanyl)-acetate
[0213] 99
[0214] Following a procedure described in Tetrahedron (1989), 45,
69, a 125 mL three-neck flask equiped with an addition funnel,
condenser and a magnetic stirrer was charged with 25 mL of
anhydrous 1,4-dioxane and 0.05 g of dirhodium diacetate. This was
refluxed under nitrogen and a solution of 2.0 g (17.5 mmol) of
ethyl diazoacetate in 20 mL of anhydrous 1,4-dioxane was added
dropwise over a period of 130 minutes. After addition was complete,
the reaction was allowed to cool to 25.degree. C. and filtered
through a short pad of alumina and concentrated under vacuum. The
residue was vacuum distilled (short path head) and the the fraction
having a bp of 61.degree.-68.degree. C. at 0.5 mm Hg was collected,
giving 1.5 g of the product as a colorless oil.
PREPARATIVE EXAMPLE 15
Ethyl tetrahydrofuran-2-acetate
[0215] 100
[0216] Following the procedure of Preparative Example 14, 2.0 g
(17.5 mmol) of ethyl diazo acetate was reacted with tetrahydrofuran
to give 1.7 g of the product as a colorless oil, bp
84.degree.-86.degree. C. at 20 mm Hg.
PREPARATIVE EXAMPLE 16
Ethyl tetrahydropyran-2-acetate
[0217] 101
[0218] Following the procedure of Preparative Example 14, 2.0 g
(17.5 mmol) of ethyl diazo acetate was reacted with tetrahydropyran
to give 1.75 g of the product as a colorless oil, bp
95.degree.-106.degree. C. at 20 mm Hg.
PREPARATIVE EXAMPLE 17
Ethyl 2-oxabicyclo[4.1.0]heptane-7-exo-acetate (18.01 and Ethyl
2-oxabicyclo[4.1.0]heptane-7-endo-acetate (19.0)
[0219] 102
[0220] Following a procedure described in Comp. Rend. (1957), 244,
2806, a 100 mL three-neck flask equiped with an addition funnel,
condenser and a magnetic stirrer was charged with 27.37 g (300
mmole) of 3,4-dihydro-2H-pyran and 0.08 g of anhydrous copper II
sulfate. This was refluxed under nitrogen and a solution of 11.42 g
(100 mmole) of ethyl diazo-acetate and 8.41 g (100 mmol) of
3,4-dihydro-2H-pyran was added dropwise over a 60 minute period.
After addition was complete, the reaction was refluxed for an
additional 2 hours and then allowed to cool to 25.degree. C. This
mixture was filtered through a short pad of alumina and
concentrated under vacuum. The residue was flash chromatographed on
silica gel using hexane-ethyl acetate (60-40) giving 10 g of the
product as a colorless oil. Silca gel TLC Rf= 0.48 using the above
chromatography solvent. NMR shows a mixture of 18.0 and 19.0 in an
15% to 85% ratio.
PREPARATIVE EXAMPLE 18
Ethyl 3-oxabicyclo[3.1.0]hexane-6-exo-acetate [20.0] and Ethyl
3-oxabicyclo[3.1.0]hexane-6-endo-acetate (21.0)
[0221] 103
[0222] Following the procedure of Preparative Example 17, react
11.42 g (100 mmole) of ethyl diazo acetate with 2,5-dihydrofuran to
give 4 g of the product as a colorless oil. Silica gel TLC Rf= 0.85
(hexane-ethyl acetate 60-40).
PREPARATIVE EXAMPLE 19
Ethyl 2-oxabicyclo[3.1.0]hexane-6-exo-acetate (22.0) and Ethyl
2-oxabicyclo[3.1.0]hexane-6-endo-acetate (23.0)
[0223] 104
[0224] Following the procedure of Preparative Example 17, react
11.42 g (100 mmole) of ethyl diazo acetate with 2,3-dihydrofuran to
give 10.4 g of the product as a colorless oil. Silica gel TLC Rf=
0.91 (hexane-ethyl acetate 60-40).
PREPARATIVE EXAMPLE 20
Ethyl 4-H-pyran-4-ylidenylacetate
[0225] 105
[0226] Following the procedure of Preparative Example 10 but using
5 g (52 mmol) of 4-H-pyran-4-one instead of tetrahydropyran-4-one,
obtain 0.4 g of the product as a yellow solid, mp= 116.5-118.7,
after flash silica gel chromatography using ethyl acetate-hexane
20%-80%.
PREPARATIVE EXAMPLE 21
Ethyl tetrahydropyran-3-acetate
[0227] 106
[0228] Following a procedure described in Comp. Rend. (1957), 244,
2806, if one were to hydrogenate the products of Preparative
Example 17 at 750 psi and 100.degree. C. using Raney nickle as the
catalyst then one would obtain the product.
PREPARATIVE EXAMPLE 22
Ethyl tetrahydrofuran-3-acetate
[0229] 107
[0230] Following a procedure described in Comp. Rend. (1957), 244,
2806, if one were to hydrogenate the products of Preparative
Example 19 at 750 psi and 100.degree. C. using Raney nickle as the
catalyst then one would obtain the product.
PREPARATIVE EXAMPLE 23
Ethyl 2,6-dimethyltetrahydropyran-4-ylidenylacetate
[0231] 108
[0232] Following the procedure of Preparative Example 10, if one
were to react 2,6-dimethyltetrahydro-4H-pyran-4-one (Recueil.
(1959) 78, 91) with sodium hydride and triethyl phosphonoacetate to
then one would obtain the product.
PREPARATIVE EXAMPLE 24
Ethyl 2,6-dimethyltetrabydropyran-4-acetate
[0233] 109
[0234] Following the procedure of Preparative Example 11, if one
were to hydrogenate the product of Preparative Example 23 one would
obtain the product.
PREPARATIVE EXAMPLE 25
Ethyl 2,2,6,6-tetramethyltetrahydropyran-4-ylidenylacetate
[0235] 110
[0236] Following the procedure of Preparative Example 10, if one
were to react 2,2,6,6-tetramethyltetrahydro-4H-pyran-4-one (J.
Chem. Soc. (1944) 338) with sodium hydride and triethyl
phosphonoacetate then one would obtain the product.
PREPARATIVE EXAMPLE 26
Ethyl 2,2,6,6-tetramethyltetrahydropvran-4-acetate
[0237] 111
[0238] Following the procedure of Preparative Example 11, if one
were to hydrogenate the product of Preparative Example 25 one would
obtain to the product.
PREPARATIVE EXAMPLE 27
Ethyl tetrahydropyran-4-acetate
[0239] 112
[0240] Following a procedure described in Liebigs Ann. Chem. (1982)
250, if one were to react ethyl tetrahydro-4-ylidenylcarboxylate,
product 15.0 of Preparative Example 10, with an excess of sodium
cyanide at 80-100.degree. C. one would obtain the product.
PREPARATIVE EXAMPLE 28
Ethyl 8-oxabicyclo[3.2.1]octa-6-ene-3-ylidenylacetate
[0241] 113
[0242] Following the procedure of Preparative Example 10, if one
were to react 8-Oxabicyclo[3.2.1]octa-6-ene-3-one (J. Am. Chem.
Soc. (1978) 100,1765) with sodium hydride and triethyl
phosphonoacetate one would obtain the product.
PREPARATIVE EXAMPLE 29
Ethyl 8-oxabicyclo[3.2.1]octa-6-ene-3-.beta.-acetate (24.0)
and
Ethyl 8-oxabicyclo[3.2.1]octa-6-ene-3-.alpha.-acetate (25.0)
[0243] 114
[0244] Following the procedure of Preparative Example 11, if one
were to hydrogenate the product of Preparative Example 28 one would
obtain the products after separation by silica gel
chromatography.
PREPARATIVE EXAMPLE 30
Ethyl 2-ethoxytetrahydropyran-3-acetate
[0245] 115
[0246] Following a procedure described in Comp. Rend. (1957), 244,
2806, if the products of Preparative Example 9 were to be reacted
with boiling ethanol containing 1-2% HCl gas then the product would
be obtained.
PREPARATIVE EXAMPLE 31
Ethyl 2-ethoxytetrahydrofuran-3-acetate
[0247] 116
[0248] Following a procedure described in Comp. Rend. (1957), 244,
2806, if the products of Preparative Example 14 were to be reacted
with boiling ethanol containing 1-2% HCl gas then the product would
be obtained.
PREPARATIVE EXAMPLE 32
Tetrahydropyran-4-acetic acid
[0249] 117
[0250] The product of Preparative Example 11 (3.04 g, 17.7 mmol)
was dissoloved in 90 mL of ethanol containing 3 g (53 mmol) of
potassium hydroxide. This was stirred for 18 hours and then
concentrated under vacuum. The residue was dissolved in 15 mL of
water, adjusted to pH 2 with 12 N HCl, and extracted with three 50
mL portions of dichloromethane. The combined organic layers were
dried over magnesium sulfate and concentrated under vacuum giving
2.04 g of the product as a white solid, mp=60-63.degree. C.
[0251] Using the hydrolysis procedure of Preparative Example 32,
the esters of Preparative Examples 10-20 were hydrolyzed to the
carboxylic acids identified as Preparative Examples 33 to 46 in
Table 1. If one were to follow the hydrolysis procedure of
Preparative Example 32, the esters of Preparative Examples 21 to 31
could be hydrolyzed to obtain the carboxylic acids identified as
Preparative Examples 47-59 in Table 1.
1TABLE 1 Starting Material (Ester) Product (Carboxylic Acid) 118
119 Preparative Example 10 Preparative Example 33 120 121
Preparative Example 10 Preparative Example 34 122 123 Preparative
Example 12 Preparative Example 35 124 125 Preparative Example 13
Preparative Example 36 126 127 Preparative Exampel 14 Preparative
Example 37 128 129 Preparative Example 15 Preparative Example 38
130 131 Preparative Example 16 Preparative Example 39 132 133
Preparative Example 17 Preparative Example 40 134 135 Preparative
Example 17 Preparative Example 41 136 137 Preparative Example 18
Preparative Example 42 138 139 Preparative Example 18 Preparative
Example 43 140 141 Preparative Example 19 Preparative Example 44
142 143 Preparative Example 19 Preparative Example 45 144 145
Preparative Example 20 Preparative Example 46 146 147 Preparative
Example 21 Preparative Example 47 148 149 Preparative Example 22
Preparative Example 48 150 151 Preparative Example 23 Preparative
Example 49 152 153 Preparative Example 24 Preparative Example 50
154 155 Preparative Example 25 Preparative Example 51 156 157
Preparative Example 26 Preparative Example 52 158 159 Preparative
Example 27 Preparative Example 53 160 161 Preparative Example 27
Preparative Example 54 162 163 Preparative Example 28 Preparative
Example 55 164 165 Preparative Example 29 Preparative Example 56
166 167 Preparative Example 29 Preparative Example 57 168 169
Preparative Example 30 Preparative Example 58 170 171 Preparative
Example 31 Preparative Example 59
PREPARATIVE EXAMPLE 60
2-Oxabicyclo[2.2.2]-5-anti-carboxylic acid
[0252] 172
[0253] Ethyl 2-oxabicyclo[2.2.2]-5-anti-carboxylate (a by-product
produced along with
5-anti-carbomethoxy-7-anti-acetoxy-2-oxabicyclo[2.2.2]octane
described in Tet. Lett. (1979) 35, 3275) was hydrolyzed following
the procedure of Preparative Example 32 to give the product as a
waxy solid.
EXAMPLE 1
(+)
-4-(3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cyclohepta[1,2-b]p-
yridine-11(R)-yl-1-[(tetrahydro-4H-pyran-4-yl)acetyl piperidine
[0254] 173
[0255] Dissolve the (+) product of Preparative Example 6, Step B,
(0.1 g, 0.212 mmol) in 5 mL of DMF, stir at room temperature and
add 0.043 g (0.424 mmol) of 4-methylmorpholine, 0.053 g (0.0276
mmol) of DEC, 0.037 g (0.276 mmol) of HOBT and 0.0397 g (0.276
mmole) of the product of Preparative Example 32. Stir the mixture
at room temperature for 18 hours, then concentrate in vacuo to a
residue and partition between methylene chloride and water. Wash
the organic phase with aqueous sodium bicarbonate solution then
brine. Dry the organic phase over magnesium sulfate, filter and
concentrate in vacuo to a residue. Chromatograph the residue on a
silica gel plate, eluting with methylene chloride-methanol (96%-4%)
to yield the product (0.13 g) as a white solid.
M.p.=83.2-88.7.degree. C., Mass Spec.: MH+=597.
[.alpha.].sub.D.sup.23.2.- degree. C.=+55.5.degree., c=0.2,
methylene chloride.
[0256] Using the coupling procedure of Example 1, the acids of
Preparative Examples 33-60 are reacted with the (+) product of
Preparative Example 6, Step B, to produce the compounds of Formula
1.16: 174
[0257] wherein R.sup.12 is as defined in Table 2 below. In Table 2,
"EX" stands for Example, and "mp" stands for melting point:
2 TABLE 2 EX R.sup.12 mp (.degree. C.) 1 175 83.2-88.7 2 176
103.3-107.9 3 177 110.3-113.9 4 178 -- 5 179 115.9-119.9 6 180
111-124 (d) 7 181 107-115 (d) 8 182 116-122 (d) 9 183 125.8-127.3
10 184 -- 11 185 124.9-127.8 12 186 -- 13 187 124.3-125.3 14 188 --
15 189 174.3-178.8 16 190 -- 17 191 -- 18 192 -- 19 193 -- 20 194
-- 21 195 -- 22 196 -- 23 197 -- 24 198 -- 25 199 -- 26 200 -- 27
201 -- 28 202 -- 29 203 152-164 (d) Isomer 1 30 204 151-159 (d)
Isomer 2 31 205 118.1-122.3 Starting acid is commerically available
32 206 -- Starting acid described in J. Am. Chem. Soc (1995) 115,
8401
EXAMPLE 33
[0258] 207
[0259] Following the procedure of Example 1, react the R-(+)-isomer
of Preparative Example 9 with the product of Preparative Example 32
to give the product as a white solid mp=93.5-97.6.degree. C.
EXAMPLE 34
[0260] 208
[0261] If the the coupling procedure described in Example 1 were to
be used, the product of Example 23 could be reacted with ammonium
chloride to produce the product.
EXAMPLE 35
[0262] 209
[0263] Dissolve 90 mg (0.14 mmol) of the product of Example 5 in 5
mL of THF and 34 mL of trifluoroacetic acid. Add 37 mL of 30%
hydrogen peroxide and stir for three days. Concentrate under vacuum
and partition the residue between water and dichloromethane. Dry
the organic layer over magnesium sulfate, concentrate under vacuum
and chromatograph the residue by preparative silica gel TLC using
dichloromethane saturated with ammonia to give the product as a
white solid. M.p.=135.6-140.degree. C., Mass Spec.: MH+=628.
EXAMPLE 36
[0264] 210
[0265] Dissolve the (+) product of Preparative Example 6, Step B
(0.5 g, 1.06 mmol) in 10 mL of dichloromethane, stir at room
temperature and add 0.128 g (1.27 mmol) of 4-methylmorpholine,
0.285 g (1.48 mmol) of DEC, 0.172 g (1.27 mmol) of HOBT and 0.097 g
(1.27 mmole) of glycolic acid. Stir the mixture at room temperature
for 18 hours, then concentrate in vacuo to a residue and partition
between methylene chloride and water. Wash the organic phase with
aqueous sodium bicarbonate solution then brine. Dry the organic
phase over magnesium sulfate, filter and concentrate in vacuo to a
residue. Chromatograph the residue by preparative silica gel TLC,
eluting with methylene chloride-methanol (95%-5%) to yield the
amide of glycolic acid and the starting tricyclic reactant of
Preparative Example 6.
[0266] Step B
[0267] Dissolve 0.34 g (0.643 mmol) of the product of Step A in 1
mL of dichloromethane containing 5.4 mL of thionyl chloride. Allow
to stir for 18 hours and concentrate under vacuum. Add 10 mL of
toluene to the residue and concentrate under vacuum and repeat this
step two additional times to give the product.
[0268] Step C
[0269] Dissolve the product of Step B in 1.0 mL of dichloromethane
followed by 0.124 g of morpholine. Stir for 18 hours then
concentrate under vacuum. Partition the residue between
dichloromethane and aqueous sodium bicarbonate solution.
Concentrate the organic layer under vacuum and chromatograph the
residue by preparative silica gel TLC using methylene
chloride-methanol (95%-5%) to yield the product as a white solid.
M.p=112.4-113.5.degree. C., Mass. Spec.: MH+=599.
EXAMPLE 37
[0270] 211
[0271] Following the procedure of Example 36, thiomorpholine was
used instead of morpholine in Step C to yield the product as a
white solid.
EXAMPLE 38
[0272] 212
[0273] Following the procedure of Example 36, except in Step A the
(-) product of Preparative Example 4 Step D was used instead of the
(+) product of Preparative Example 6, Step B, and thiomorpholine
was used instead of morpholine in Step C, to yield the product as a
white solid.
EXAMPLE 39
[0274] 213
[0275] The product of Example 38 was reacted under the conditions
of Example 35 to yield the product as a white solid.
EXAMPLE 40
[0276] 214
[0277] Dissolve 160 mg (0.268 mmol) of the product of Example 1 in
3 mL of CH.sub.2Cl.sub.2 and add 162.3 mg (0.536 mmol) of
4-chloroperoxybenzoic acid (57% pure) and stir for 3 hr. Dilute
with 50 mL of CH.sub.2Cl.sub.2 then wash with saturated NaHCO.sub.3
followed by brine. Dry the organic layer over MgSO.sub.4,
concentrate in vacuo and purify the residue by preparative silica
gel TLC using 2% methanol in CH.sub.2Cl.sub.2 saturated with
ammonia to give 116 mg (71%) of the title compound as a white
solid. m.p.=141-151.degree. C. (dec); MS MH+= 613.
ASSAYS
[0278] FPT IC.sub.50 (inhibition of famesyl protein transferase, in
vitro enzyme assay) and COS Cell IC.sub.50 (Cell-Based Assay) were
determined following the assay procedures described in WO 95/10516,
published Apr. 20, 1995. GGPIT IC.sub.50 (inhibition of
geranylgeranyl protein transferase, in vitro enzyme assay), Cell
Mat Assay, and anti-tumor activity (in vivo anti-tumor studies)
could be determined by the assay procedures described in WO
95/10516. The disclosure of WO 95/10516 is incorporated herein by
reference thereto.
[0279] Additional assays can be carried out by following
essentially the same procedure as described above, but with
substitution of alternative indicator tumor cell lines in place of
the T24-BAG cells. The assays can be conducted using either
DLD-1-BAG human colon carcinoma cells expressing an activated K-ras
gene or SW620-BAG human colon carcinoma cells expressing an
activated K-ras gene. Using other tumor cell lines known in the
art, the activity of the compounds of this invention against other
types of cancer cells could be demonstrated.
[0280] Soft Agar Assay:
[0281] Anchorage-independent growth is a characteristic of
tumorigenic cell lines. Human tumor cells can be suspended in
growth medium containing 0.3% agarose and an indicated
concentration of a farnesyl transferase inhibitor. The solution can
be overlayed onto growth medium solidified with 0.6% agarose
containing the same concentration of farnesyl transferase inhibitor
as the top layer. After the top layer is solidified, plates can be
incubated for 10-16 days at 37.degree. C. under 5% CO.sub.2 to
allow colony outgrowth. After incubation, the colonies can be
stained by overlaying the agar with a solution of MTT (3-[
4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide,
Thiazolyl blue) (1 mg/mL in PBS). Colonies can be counted and the
IC.sub.50's can be determined.
[0282] The results are given in Table 3. In Table 3, "nM"
represents nanomolar.
3TABLE 3 Compound of FPT IC.sub.50 COS Cell IC.sub.50 Example No.
(nM) (nM) 1 0.4 1.8 1.2 12 2.4 15 2 5.5 285 3 6.1 -- 5 3.5 143 6 23
-- 7 42 -- 8 2.0 68 29 6.7 30 30 7.5 75 33 45 -- 35 2.3 -- 36 3.1
-- 37 4.9 <10 39 4.7 68 40 2.2 100% @ 10
[0283] The compound of Example 40 had a Soft Agar IC.sub.50 of 8
nM.
[0284] For preparing pharmaceutical compositions from the compounds
described by this invention, inert, pharmaceutically acceptable
carriers can be either solid or liquid. Solid form preparations
include powders, tablets, dispersible granules. capsules, cachets
and suppositories. The powders and tablets may be comprised of from
about 5 to about 70 percent active ingredient. Suitable solid
carriers are known in the art, e.g. magnesium carbonate, magnesium
stearate, talc, sugar, lactose. Tablets, powders, cachets and
capsules can be used as solid dosage forms suitable for oral
administration.
[0285] For preparing suppositories, a low melting wax such as a
mixture of fatty acid glycerides or cocoa butter is first melted,
and the active ingredient is dispersed homogeneously therein as by
stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool and thereby solidify.
[0286] Liquid form preparations include solutions, suspensions and
emulsions. As an example may be mentioned water or water-propylene
glycol solutions for parenteral injection.
[0287] Liquid form preparations may also include solutions for
intranasal administration.
[0288] Aerosol preparations suitable for inhalation may include
solutions and solids in powder form, which may be in combination
with a pharmaceutically acceptable carrier, such as an inert
compressed gas.
[0289] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for either oral or parenteral administration. Such liquid forms
include solutions, suspensions and emulsions.
[0290] The compounds of the invention may also be deliverable
transdermally. The transdermal compositions can take the form of
creams, lotions, aerosols and/or emulsions and can be included in a
transdermal patch of the matrix or reservoir type as are
conventional in the art for this purpose.
[0291] Preferably the compound Is administered orally.
[0292] Preferably, the pharmaceutical preparation is in unit dosage
form. In such form, the preparation is subdivided into unit doses
containing appropriate quantities of the active component, e.g., an
effective amount to achieve the desired purpose.
[0293] The quantity of active compound in a unit dose of
preparation may be varied or adjusted from about 0.1 mg to 1000 mg,
more preferably from about 1 mg. to 300 mg, according to the
particular application.
[0294] The actual dosage employed may be varied depending upon the
requirements of the patient and the severity of the condition being
treated. Determination of the proper dosage for a particular
situation is within the skill of the art. Generally, treatment is
initiated with smaller dosages which are less than the optimum dose
of the compound. Thereafter, the dosage is increased by small
increments until the optimum effect under the circumstances is
reached. For convenience, the total daily dosage may be divided and
administered in portions during the day if desired.
[0295] The amount and frequency of administration of the compounds
of the invention and the pharmaceutically acceptable salts thereof
will be regulated according to the judgment of the attending
clinician considering such factors as age, condition and size of
the patient as well as severity of the symptoms being treated. A
typical recommended dosage regimen is oral administration of from
10 mg to 2000 mg/day preferably 10 to 1000 mg/day, in two to four
divided doses to block tumor growth. The compounds are non-toxic
when administered within this dosage range.
[0296] The following are examples of pharmaceutical dosage forms
which contain a compound of the invention. The scope of the
invention in its pharmaceutical composition aspect is not to be
limited by the examples provided.
Pharmaceutical Dosage Form Examples
EXAMPLE A
[0297]
4 Tablets No. Ingredients mg/tablet mg/tablet 1. Active compound
100 500 2. Lactose USP 122 113 3. Corn Starch, Food Grade, 30 40 as
a 10% paste in Purified Water 4. Corn Starch, Food Grade 45 40 5.
Magnesium Stearate 3 7 Total 300 700
Method of Manufacture
[0298] Mix Item Nos. 1 and 2 in a suitable mixer for 10-15 minutes.
Granulate the mixture with Item No. 3. Mill the damp granules
through a coarse screen (e.g., 1/4", 0.63 cm) if necessary. Dry the
damp granules. Screen the dried granules if necessary and mix with
Item No. 4 and mix for 10-15 minutes. Add Item No. 5 and mix for
1-3 minutes. Compress the mixture to appropriate size and weigh on
a suitable tablet machine.
EXAMPLE B
[0299]
5 Capsules No. Ingredient mg/capsule mg/capsule 1. Active compound
100 500 2. Lactose USP 106 123 3. Corn Starch, Food Grade 40 70 4.
Magnesium Stearate NF 7 7 Total 253 700
Method of Manufacture
[0300] Mix Item Nos. 1, 2 and 3 in a suitable blender for 10-15
minutes. Add Item No. 4 and mix for 1-3 minutes. Fill the mixture
into suitable two-piece hard gelatin capsules on a suitable
encapsulating machine.
[0301] While the present invention has been described in
conjunction with the specific embodiments set forth above, many
alternatives, modifications and variations thereof will be apparent
to those of ordinary skill in the art. All such alternatives,
modifications and variations are intended to fall within the spirit
and scope of the present invention.
* * * * *