U.S. patent application number 10/454960 was filed with the patent office on 2004-02-26 for 6,7-dimethoxyquinazolines and therapeutic use thereof.
This patent application is currently assigned to Parker Hughes Institute. Invention is credited to Liu, Xing-Ping, Narla, Rama Krishna, Uckun, Fatih M..
Application Number | 20040039002 10/454960 |
Document ID | / |
Family ID | 27494574 |
Filed Date | 2004-02-26 |
United States Patent
Application |
20040039002 |
Kind Code |
A1 |
Uckun, Fatih M. ; et
al. |
February 26, 2004 |
6,7-Dimethoxyquinazolines and therapeutic use thereof
Abstract
Quinazoline compounds and methods for the treatment of cancer
and for the treatment of allergic reactions.
Inventors: |
Uckun, Fatih M.; (White Bear
Lake, MN) ; Liu, Xing-Ping; (Minneapolis, MN)
; Narla, Rama Krishna; (St. Paul, MN) |
Correspondence
Address: |
Attention: Anna M. Nelson
MERCHANT & GOULD P.C.
P.O. Box 2903
Minneapolis
MN
55402-0903
US
|
Assignee: |
Parker Hughes Institute
St. Paul
MN
|
Family ID: |
27494574 |
Appl. No.: |
10/454960 |
Filed: |
June 5, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10454960 |
Jun 5, 2003 |
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09923903 |
Aug 7, 2001 |
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09923903 |
Aug 7, 2001 |
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09779809 |
Feb 8, 2001 |
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6358962 |
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09779809 |
Feb 8, 2001 |
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09357404 |
Jul 20, 1999 |
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6258820 |
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60125177 |
Mar 19, 1999 |
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60125338 |
Mar 19, 1999 |
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60125145 |
Mar 19, 1999 |
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Current U.S.
Class: |
514/266.4 ;
514/267; 544/293 |
Current CPC
Class: |
A61P 35/02 20180101;
A61P 35/00 20180101; C07D 239/94 20130101; A61P 37/00 20180101 |
Class at
Publication: |
514/266.4 ;
544/293; 514/267 |
International
Class: |
A61K 031/517; C07D
239/72 |
Claims
We claim:
1. A compound of the formula: 93wherein: R.sup.a is iodo,
(C.sub.1-C.sub.4)hydroxyalkyl, methylenedioxy, ethylenedioxy,
benzyloxy, OCF.sub.3. SCF.sub.3SO.sub.3H, SO.sub.2F,
SO.sub.2NR.sup.2R.sup.3 where R.sup.2 is hydrogen or
(C.sub.1-C.sub.4)alkyl and R.sup.3 is hydrogen,
(C.sub.1-C.sub.4)alkyl, or phenyl, NR.sup.2R.sup.4 where R.sup.2 is
hydrogen or (C.sub.1-C.sub.4)alkyl and R.sup.4 is phenyl; or a
group of the formula 94wherein R.sup.5 and R.sup.6 are each
independently, hydrogen, (C.sub.1-C.sub.4)alkyl, or
(C.sub.1-C.sub.4) perfluoroalkyl, and R.sup.7 is hydrogen, halo,
hydroxy, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)hydroxyalkyl, or N(R.sup.2).sub.2, where R.sup.2
is hydrogen or (C.sub.1-C.sub.4)alkyl; n is an integer of 1-4;
R.sup.b is each, independently, hydrogen, halo, hydroxy, mercapto,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)thioalkyl, (C.sub.1-C.sub.4)hydroxyalkyl, nitro,
cyano, methylenedioxy, ethylenedioxy, COCH.sub.3, CF.sub.3,
OCF.sub.3, SCF.sub.3, COOH, SO.sub.3H, SO.sub.2F, phenyl, or phenyl
substituted by a group selected from halo, hydroxy, mercapto,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4) alkoxy,
(C.sub.1-C.sub.4)thioalkyl, (C.sub.1-C.sub.4)hydroxyalkyl, amino,
nitro, cyano, CF.sub.3, COOH, SO.sub.3H, SO.sub.2NR.sup.2R.sup.3,
SO.sub.2F where R.sup.2 is H or (C.sub.1-C.sub.4)alkyl and R.sup.3
is H, (C.sub.1-C.sub.4)alkyl, phenyl, or phenyl substituted by a
group as defined above; benzyloxy or benzyloxy substituted on the
phenyl portion by a group defined above; NR.sup.2R.sup.3 where
R.sup.2 is H or (C.sub.1-C.sub.4)alkyl and R.sup.3 is H,
(C.sub.1-C.sub.4)alkyl, phenyl, or phenyl substituted by a group as
defined above; and R.sup.1 is (C.sub.1-C.sub.4)alkyl or a
pharmaceutically acceptable salt thereof.
2. The compound of claim 1 wherein the pharmaceutically acceptable
salt is an acid addition salt.
3. The compound of claim 1, wherein R.sup.1 is methyl.
4. The compound of claim 3, wherein R.sup.a is a member selected
from the group consisting of I, NHC.sub.6H.sub.5,
--OCH.sub.2CH.sub.2O--, --OCH.sub.2O--, OCF.sub.3, SCF.sub.3,
CH.sub.2OH, C.sub.2H.sub.4OH, SO.sub.3H, SO.sub.2NH.sub.2, and
SO.sub.2F.
5. The compound of claim 4, wherein R.sup.a is I, OCF.sub.3, or
SO.sub.2F.
6. The compound of claim 5, wherein R.sup.a is I.
7. The compound of claim 5 wherein R.sup.a is OCF.sub.3.
8. The compound of claim 3, wherein n is I and R.sup.a is a group
of the formula 95
9. The compound of claim 8, wherein R.sup.5 and R.sup.6 are each,
independently, H, CH.sub.3, or CF.sub.3.
10. The compound of claim 9, wherein R.sup.5 and R.sup.6 are
CF.sub.3 and R.sup.7 is NH.sub.2.
11. The compound of claim 3, wherein R.sup.b is at least one member
selected from the group consisting of F, Cl, Br, I, OH, NH.sub.2,
NO.sub.2, CN, COOH, CH.sub.3 and CF.sub.3.
12. The compound of claim 11, wherein R.sup.b is at least one
member selected from the group consisting of F, Cl, Br, OH and
CF.sub.3.
13. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1 and a pharmaceutically
acceptable carrier or diluent.
14. A method of treating tumors in a subject comprising
administering to said subject an anti-tumor effective amount of a
compound of claim 1.
15. A method of treating leukemia in a subject comprising
administering to said subject a compound of claim 1 in an amount
effective for inducing apoptosis of leukemia cells.
16. A method of inhibiting cancer cell growth in a subject
comprising administering to said subject a cytotoxic effective
amount of a compound of claim 1.
17. A method of treating leukemia in a subject comprising
administering to said subject a compound selected from the group
consisting of:
4-(3',5'-dibromo-4'-methylphenyl)amino-6,7-dimethoxyquinazoline,
4-(2',4',6'-tribromophenyl)amino-6,7-dimethoxyquinazoline,
4-(2',3',5',6'-tetrafluoro-4'-bromophenyl)amino-6,7-dimethoxyquinazoline,
4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(3'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(2'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(4'-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
4-(2'-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
and pharmaceutically acceptable acid addition salts thereof.
18. A method of treating breast tumors in a subject comprising
administering to said subject a compound selected from the group
consisting of 4-(3'-bromophenyl)amino-6,7-dimethoxyquinazoline,
4-(3',5'-dibromo-4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,
4-(3'-chloro-4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
4-(2',3',5',6'-tetrafluoro-4'-bromophenyl)amino-6,7-dimethoxyquinazoline,
4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(3'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(2'-fluorophenyl)amino-6,7-dimethoxyquinazoline, and
pharmaceutically acceptable acid addition salts thereof.
19. A method of treating multi-drug resistant leukemia in a subject
comprising administering to said subject
4-(4'-hydroxyphenyl)amino-6,7-di- methoxyquinazoline or a
pharmaceutically acceptable salt thereof, in an amount effective
for inducing apoptosis of multi-drug resistant leukemia cells.
20. A method of treating multi-drug resistant leukemia in a subject
comprising administering to said subject
4-(3'-bromo-4'-hydroxyphenyl)ami- no-6,7-dimethoxyquinazoline or a
pharmaceutically acceptable acid addition salt thereof, in an
amount effective for inducing apoptosis of multi-drug resistant
leukemia cells.
21. The compound
4-(3',5'-dibromo-4'-methylphenyl)amino-6,7-dimethoxyquina- zoline
or a pharmaceutically acceptable acid addition salt thereof.
22. The compound
4-2',4',6'-tribromophenyl)amino6,7-dimethoxyquinazoline or a
pharmaceutically acceptable acid addition salt thereof.
23. The compound
4-(2',3',5',6'-tetrafluoro-5'-bromophenyl)amino-6,7-dimet-
hoxyquinazoline or a pharmaceutically acceptable acid addition salt
thereof.
24. The compound 4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline
or a pharmaceutically acceptable acid addition salt thereof.
25. The compound
4-(4'-trifluoromethylphenyl)amino-6,7-dimethoxyquinazolin- e or a
pharmaceutically acceptable acid addition salt thereof.
26. The compound
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxy-qu-
inazoline or a pharmaceutically acceptable acid addition salt
thereof.
27. A pharmaceutical composition comprising an effective amount of
a compound selected from the group consisting of:
4-(3,5-bis-trifluoromethy- lphenyl)amino-6,7-dimethoxyquinazoline,
4-(2',4',6'-tribromophenyl)amino-6- ,7-dimethoxyquinazoline,
4-(2',3',5',6'-tetrafluoro-4'-bromophenyl)amino-6-
,7-dimethoxyquinazoline,
4-(3',5'-dibromo-4'-methylphenyl)amino-6,7-dimeth- oxyquinazoline,
4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
4-(4'-tifluoromethylphenyl)amino-6,7-dimethoxyquinazoline, and
pharmaceutically acceptable acid addition salts thereof; and a
pharmaceutically acceptable carrier or diluent.
Description
FIELD OF THE INVENTION
[0001] This application relates to quinazoline compounds,
compositions and therapeutic methods for the treatment of cancers
and treatment of allergic disorders by administering quinazoline
compounds.
BACKGROUND OF THE INVENTION
[0002] Quinazoline compounds have been suggested as useful
compounds in the treatment of cell growth and differentiation
characterized by activity of the human epidermal growth factor
receptor type2 (HER2). See, for example, Myers et al., U.S. Pat.
No. 5,721,237. Some quinazoline derivatives have been suggested as
useful as anti cancer agents for the treatment of specific receptor
tyrosine kinase-expressing cancers, especially those expressing
epithelial growth factor (EGF) receptor tyrosine kinase. See, for
example, Barker et al., U.S. Pat. No. 5,457,105. It is generally
taught that quinazolines exert their anti-tumor effects via
tyrosine kinase inhibition. However, while some quinazoline
compounds inhibit the growth of brain tumor cells, others with
equally potent tyrosine kinase inhibitory activity fail to do so
(Naria et.al., 1998, Clin Cancer Res. 4:1405-1414; Naria et al.,
1998, Clin Cancer Res. 4:2463-2471).
[0003] Several tumors expressing EGF receptors are not killed by
quinazoline compounds, whereas some tumors not expressing EGF
recptors are. Thus, the cytotoxic activity of quinazoline compounds
cannot be attributed to the compound's tyrosine kinase inhibitory
activity, and particularly not to the compound's ability to inhibit
EGF receptor tyrosine kinase. A chemical structure-activity
relationship determining the anti-cancer activity of quinazoline
derivatives has not been established.
[0004] Novel quinazoline compounds may provide potent new
therapeutic molecules for the treatment of disorders such as
cancers. Methods of using both known and novel quinazoline
compounds that employ an understanding of structure-function
relationships are needed.
SUMMARY OF THE INVENTION
[0005] A series of quinazoline compounds were synthesized and
analyzed for therapeutic activities, including anti cancer
activities, particularly against EGR receptor-negative leukemias.
Specific quinazoline compounds of the invention were found to
possess potent and specific tyrosine kinase inhibitory activities
affecting cell proliferation and survival. Quinazoline compounds of
the invention are demonstrated as useful for the treatment of
specific tumors, including breast tumors, brain tumors, and
leukemias, particularly EGF receptor-negative leukemias, and to be
particularly useful in the treatment of multi rug resistant
leukemias.
[0006] The invention provides novel quinazoline compounds of
formula I, as disclosed below, as well as therapeutic methods
utilizing these compounds.
BRIEF DESCRIPTION OF THE DRAWINGS
[0007] FIGS. 1A-1C are graphs showing cytotoxic activity of
fluoro-substituted dimethoxy quinazoline compounds (F-dmQ) against
leukemic NALM-6 cells.
[0008] FIGS. 2A-2C are graphs showing cytotoxic activity of F-dmQ
on breast cancer BT-20 cells.
[0009] FIGS. 3A-3F are photographs showing induction of apoptosis
in cancer cells by F-dmQ.
[0010] FIG. 4 is a bar graph showing adhesive properties of various
cancer cells to extracellular matrix proteins.
[0011] FIGS. 5A-5F are bar graphs showing the effect of F-dmQ on
cancer cell-adhesion to extracellular matrix (ECM) proteins.
[0012] FIGS. 6A-6E are photographs showing the effects of HI-P353
and HI-P364 on glioblastoma cell migration from spheroids.
[0013] FIGS. 7A and 7B are bar graphs showing the anti-invasive
activity of fluoro-substituted quinazoline compounds (F-dmQ)
against glioblastoma U373 and breast cancer MDA-MB-231 cells.
[0014] FIGS. 8A-8D are photographs showing depolymerizaton of actin
stress fibers and microtubules by HI-P353.
[0015] FIGS. 9A-9I are photographs showing inhibition of actin
stress fiber formation in glioblastoma cells by HI-P154.
[0016] FIGS. 10A-10C are graphs showing the inhibition of cancer
cell growth in vivo by the quinazolines of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0017] Definitions:
[0018] The terms "quinazoline", "quinazoline compound", and
"quinazoline derivative" are used interchangeably in this
application to mean compounds of formula I. All scientific and
technical terms used in this application have meanings commonly
used in the art unless otherwise specified. As used in this
application, the following words or phrases have the meanings
specified.
[0019] Halo is fluoro, chloro, bromo, or iodo. Alkyl, alkanoyl,
etc., denote both straight and branched groups; but reference to an
individual radical such as "propyl" embraces only the straight
chain radical, a branched chain isomer such as "isopropyl" being
specifically referred to. (C.sub.1-C.sub.4)alkyl includes methyl,
ethyl, propyl, isopropyl, butyl, iso-butyl, and sec-butyl;
(C.sub.1-C.sub.4)alkoxy includes methoxy, ethoxy, propoxy,
isopropoxy, butoxy, iso-butoxy, and sec-butoxy; and
(C.sub.1-C.sub.4)alkanoyl includes acetyl, propanoyl and
butanoyl.
[0020] As used herein, "pharmaceutically acceptable carrier" means
any material which, when combined with the compound of the
invention, allows the compound to retain biological activity, such
as the ability to potentiate antibacterial activity of mast cells
and macrophages. Examples include, but are not limited to, any of
the standard pharmaceutical carriers such as a phosphate buffered
saline solution, water, emulsions such as oil/water emulsions, and
various types of wetting agents. Compositions comprising such
carriers are formulated by well known conventional methods (see,
for example, Remington's Pharmaceutical Sciences, Chapter 43, 14th
Ed., Mack Publishing Co., Easton, Pa.).
[0021] The term "conjugate" means a compound formed as a composite
between two or more molecules. More specifically, in the present
invention, the quinazoline derivative is bonded, for example,
covalently bonded, to cell-specific targeting moieties forming a
conjugate compound for efficient and specific delivery of the agent
to a cell of interest.
[0022] The phrase "targeting moiety" means a molecule which serves
to deliver the compound of the invention to a specific site for the
desired activity. Targeting moieties include, for example,
molecules that specifically bind molecules on a specific cell
surface. Such targeting moieties useful in the invention include
anti-cell surface antigen antibodies. Cytokines, including
interleukins and factors such as granulocyte/macrophage stimulating
factor (GMCSF) are also specific targeting moieties, known to bind
to specific cells expressing high levels of their receptors.
[0023] The term "prodrug moiety" is a substitution group which
facilitates use of a compound of the invention, for example by
facilitating entry of the drug into cells or administration of the
compound. The prodrug moiety may be cleaved from the compound, for
example by cleavage enzymes in vivo. Examples of prodrug moieties
include phosphate groups, peptide linkers, and sugars, which
moieties can be hydrolyzed in vivo.
[0024] "inhibit" means to reduce by a measurable amount, or to
prevent entirely.
[0025] "to treat" means to inhibit or block at least one symptom
that characterizes a pathologic condition, in a mammal threatened
by, or afflicted with, the condition.
[0026] Compounds of the Invention
[0027] Compounds of the invention include quinazolines having the
formula: 1
[0028] where:
[0029] R.sup.8 is iodo; (C.sub.1-C.sub.4)hydroxyalkyl,
methylenedioxy, ethylenedioxy, benzyloxy, OCF.sub.3. SCF.sub.3,
SO.sub.3H, SO.sub.2F, SO.sub.2NR.sup.2R.sup.3 in which R.sup.2 is
hydrogen or (C.sub.1-.sub.4)alkyl and R.sup.3 is hydrogen,
(C.sub.1-C.sub.4)alkyl, or phenyl, NR.sup.2R.sup.4 in which R.sup.2
is as defined above and R.sup.4 is phenyl, or R.sup.a a group of
the formula 2
[0030] in which R.sup.5 and R.sup.6 are each, independently,
hydrogen, (C.sub.1-C.sub.4)alkyl or
(C.sub.1-C.sub.4)perfluoroalkyl, and R.sup.7 is hydrogen, halo,
hydroxy, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)hydroxyalkyl, or N(R.sup.2).sub.2 in which R.sup.2
is as defined above;
[0031] n is an integer of 1-4;
[0032] R.sup.b is each, independently, hydrogen; halo; hydroxy;
mercapto; (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)thioalk- yl, (C.sub.1-C.sub.4)hydroxyalkyl, nitro,
cyano, methylenedioxy, ethylenedioxy, COCH.sub.3, CF.sub.3,
OCF.sub.3; SCF.sub.3; COOH; SO.sub.3H; SO.sub.2F; phenyl or phenyl
substituted by a group selected from halo, hydroxy, mercapto,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)thioalkyl, (C.sub.1-C.sub.4)hydroxyalkyl, amino,
nitro, cyano, CF.sub.3, COOH, SO.sub.3H, SO.sub.2NR.sup.2R.sup.3 in
which R.sup.2 and R.sup.3 are as defined below, and SO.sub.2F.
R.sup.a can also be benzyloxy or benzyloxy substituted on the
phenyl portion by a group defined above, NR.sup.2R.sup.3 in which
R.sup.2 is H or (C.sub.1-C.sub.4)alkyl and R.sup.3 is H,
(C.sub.1-C.sub.4)alkyl, phenyl or phenyl substituted by a group as
defined above;
[0033] R.sup.1 is (C.sub.1-C.sub.4)alkyl, preferably methyl, or a
pharmaceutically acceptable salt thereof, such as an acid addition
salt.
[0034] Preferably, R.sup.a is a member selected from the group
consisting of I, NHC6H5, --OCH.sub.2CH.sub.2O--, --OCH.sub.2O--,
OCF.sub.3, SCF.sub.3, CH.sub.2OH, C.sub.2H40H, SO.sub.3H,
SO.sub.2NH.sub.2, and SO.sub.2F; and ore preferably R.sup.a is I
OCF.sub.3 or SO.sub.2F. Most preferably, R.sup.a is I or R.sup.a is
OCF.sub.3.
[0035] In an alternative preferred compound, n is I and R.sup.a is
a group of the formula: 3
[0036] Preferably, R.sup.5 and R.sup.6 are each, independently, H,
CH.sub.3 or CF.sub.3, and most preferably, R.sup.5 and R.sup.6 are
CF.sub.3 and R.sup.7 is NH.sub.2.
[0037] In another preferred compound, R.sup.b is at least one
member selected from the group consisting of F, Cl, Br, I, OH,
NH.sub.2, NO.sub.2, CN, COOH, CH.sub.3, and CF.sub.3, and more
preferably R.sup.b is at least one member selected from the group
consisting of F, Cl, Br, OH, and CF.sub.3.
[0038] Additional preferred quinazoline compounds useful in the
treatment of tumors are described more fully below and particularly
in the Examples. These include:
[0039]
4-(3',5'-dibromo-4'-methylphenyl)amino-6,7-dimethoxyquinazoline;
[0040]
4-(2',4',6'-tribromophenyl)amino-6,7-dimethoxyquinazoline;
[0041]
4-(2',3',5',6'-tetrafluoro-5'-bromophenyl)amino-6,7-dimethoxyquinaz-
oline;
[0042] 4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline;
[0043] 4-(4'trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline;
and
[0044]
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline.
[0045] Methods of Treatment
[0046] The compounds of the invention are useful for the treatment
of animals, including humans. In particular, the compounds of the
invention have been found to be potent inhibitors of tumor cell
proliferation and survival, and effective to induce apoptosis of
malignant cells.
[0047] Compounds of the invention have surprisingly been found to
be effective for inducing apoptosis and/or cytotoxicity of leukemia
cells. In particular,
4-(4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline compounds of the
invention have been found to effectively induce apoptosis in
multi-drug resistant leukemia. A preferred compound for the
treatment of multi-drug resistant leukemia is
4-(3'-bromo-4'-hydroxypheny- l)amino-6,7-dimethoxyquinazoline.
[0048] Compounds of the invention that are particularly useful for
treating leukemia include:
[0049]
4-(3',5'-bromo-4-methylphenyl)amino-6,7-dimethoxyquinazoline,
[0050]
4-(2',4',6'-tribromophenyl)amino-6,7-dimethoxyquinazoline,
[0051]
4-(2',3',5',6'-tetrafluoro-4'-bromophenyl)amino-6,7-dimethoxyquinaz-
oline,
[0052] 4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
[0053] 4-(3'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
[0054] 4-(2'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
[0055]
4-(4'-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
[0056] 4-(2'-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
and
[0057]
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline.
[0058] Compounds of the invention that are particularly useful for
treating breast tumors include:
[0059] 4-(3'-bromophenyl)amino-6,7-dimethoxyquinazoline,
[0060]
4-(3',5'-dibromo-4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,
[0061]
4-(3'-chloro-4'-hydroxyphenyl)amino-4,7-dimethoxyquinazoline,
[0062]
4-(3',5'-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,
[0063]
4-(2',3',5',6'-tetrafluoro-4'-bromophenyl)amino-6,7-dimethoxyquinaz-
oline,
[0064] 4-(4'-fluorophenyl)amino-6,7-dimethoxyquinazoline,
[0065] 4-(3'-fluorophenyl)amino-6,7-dimethoxyquinazoline, and
[0066] 4-(2'-fluorophenyl)amino-6,7-dimethoxyquinazoline.
[0067] Compositions
[0068] The compounds of the invention are useful as pharmaceutical
compositions prepared with a therapeutically effective amount of a
compound of claim 1 and a pharmaceutically acceptable carrier or
diluent.
[0069] The quinazoline compounds of the invention can be formulated
as pharmaceutical compositions and administered to a mammalian
host, such as a human patient in a variety of forms adapted to the
chosen route of administration, i.e., orally or parenterally, by
intravenous, intramuscular, topical or subcutaneous routes.
[0070] Thus, quinazoline compounds of the invention may be
systemically administered, e.g., orally, in combination with a
pharmaceutically acceptable vehicle such as an inert diluent or an
assimilable edible carrier, or by inhalation or insufflation. They
may be enclosed in hard or soft shell gelatin capsules, may be
compressed into tablets, or may be incorporated directly with the
food of the patient's diet. For oral therapeutic administration,
the quinazoline compounds may be combined with one or more
excipients and used in the form of ingestible tablets, buccal
tablets, troches, capsules, elixirs, suspensions, syrups, wafers,
and the like. The quinazoline compounds may be combined with a fine
inert powdered carrier and inhaled by the subject or insufflated.
Such compositions and preparations should contain at least 0.1%
quinazoline compounds. The percentage of the compositions and
preparations may, of course, be varied and may conveniently be
between about 2 to about 60% of the weight of a given unit dosage
form. The amount of quinazoline compounds in such therapeutically
useful compositions is such that an effective dosage level will be
obtained.
[0071] The tablets, troches, pills, capsules, and the like may also
contain the following: binders such as gum tragacanth, acacia, corn
starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch, alginic
acid and the like; a lubricant such as magnesium stearate; and a
sweetening agent such as sucrose, fructose, lactose or aspartame or
a flavoring agent such as peppermnint, oil of wintergreen, or
cherry flavoring may be added. When the unit dosage form is a
capsule, it may contain, in addition to materials of the above
type, a liquid carrier, such as a vegetable oil or a polyethylene
glycol. Various other materials may be present as coatings or to
otherwise modify the physical form of the solid unit dosage form.
For instance, tablets, pills, or capsules may be coated with
gelatin, wax, shellac or sugar and the like. A syrup or elixir may
contain the active compound, sucrose or fructose as a sweetening
agent, methyl and propylparabens as preservatives, a dye and
flavoring such as cherry or orange flavor. Of course, any material
used in preparing any unit dosage form should be pharmaceutically
acceptable and substantially non-toxic in the amounts employed. In
addition, the quinazoline compounds may be incorporated into
sustained-release preparations and devices.
[0072] The quinazoline compounds may also be administered
intravenously or intraperitoneally by infusion or injection.
Solutions of the quinazoline compounds can be prepared in water,
optionally mixed with a nontoxic surfactant. Dispersions can also
be prepared in glycerol liquid polyethylene glycols, triacetin and
mixtures thereof and in oils. Under ordinary conditions of storage
and use, these preparations contain a preservative to prevent the
growth of microorganisms.
[0073] The pharmaceutical dosage forms suitable for injection or
infusion can include sterile aqueous solutions or dispersions or
sterile powders comprising the quinazoline compounds which are
adapted for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions, optionally encapsulated in
liposomes. In all cases, the ultimate dosage form must be sterile,
fluid and stable under the conditions of manufacture and storage.
The liquid carrier or vehicle can be a solvent or liquid dispersion
medium comprising, for example, water, ethanol, a polyol (for
example, glycerol, propylene glycol, liquid polyethylene glycols,
and the like), vegetable oils, nontoxic glyceryl esters, and
suitable mixtures thereof. The proper fluidity can be maintained,
for example, by the formation of liposomes, by the maintenance of
the required particle size in the case of dispersions or by the use
of surfactants. The prevention of the action of microorganisms can
be brought about by various antibacterial and antifungal agents,
for example, parabens, chlorobutanol, phenol, sorbic acid,
thimerosal, and the like. In many cases, it will be preferable to
include isotonic agents, for example, sugars, buffers or sodium
chloride. Prolonged absorption of the injectable compositions can
to be brought about by the use in the compositions of agents
delaying absorption, for example, aluminum monostearate and
gelatin.
[0074] Sterile injectable solutions are prepared by incorporating
the quinazoline compounds in the required amount in the appropriate
solvent with various of the other ingredients enumerated above, as
required, followed by filter sterilization. In the case of sterile
powders for the preparation of sterile injectable solutions, the
preferred methods of preparation are vacuum drying and the freeze
drying techniques, which yield a powder of the active ingredient
plus any additional desired ingredient present in the previously
sterile-filtered solutions.
[0075] For topical administration, the quinazoline compounds may be
applied in pure form, i.e., when they are liquids. However, it will
generally be desirable to administer them to the skin as
compositions or formulations, in combination with a
dermatologically acceptable carrier, which may be a solid or a
liquid.
[0076] Useful solid carriers include finely divided solids such as
talc, clay, microcrystalline cellulose, silica, alumina and the
like. Other solid carriers include nontoxic polymeric nanoparticles
or microparticles. Useful liquid carriers include water, alcohols
or glycols or water-alcohol/glycol blends, in which the quinazoline
compounds can be dissolved or dispersed at effective levels,
optionally with the aid of non-toxic surfactants. Adjuvants such as
fragrances and additional antimicrobial agents can be added to
optimize the properties for a given use. The resultant liquid
compositions can be applied from absorbent pads, used to impregnate
bandages and other dressings, or sprayed onto the affected area
using pumppe or aerosol sprayers.
[0077] Thickeners such as synthetic polymers, fatty acids, fatty
acid salts and esters, fatty alcohols, modified celluloses or
modified mineral materials can also be employed with liquid
carriers to form spreadable pastes, gels, ointments, soaps, and the
like, for application directly to the skin of the user.
[0078] Examples of useful dermatological compositions which can be
used to deliver the quinazoline compounds to the skin are known to
the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392),
Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No.
4,559,157) and Wortztman (U.S. Pat. No. 4,820,508).
[0079] Useful dosages of the compounds of formula I can be
determined by comparing their in vitro activity, and in vivo
activity in animal models. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art; for example, see U.S. Pat. No. 4,938,949.
[0080] Generally, the concentration of the quinazoline compounds in
a liquid composition, such as a lotion, will be from about 0.1-25
wt-%, preferably from about 0.5-10 wt-%. The concentration in a
semi-solid or solid composition such as a gel or a powder will be
about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%.
[0081] The amount of the quinazoline compounds required for use in
treatment will vary not only with the particular salt selected but
also with the route of administration, the nature of the condition
being treated and the age and condition of the patient and will be
ultimately at the discretion of the attendant physician or
clinician.
[0082] In general, however, a suitable dose will be in the range of
from abut 0.5 to about 100 mg/kg, e.g., from about 10 to about 75
mg/kg of body weight per day, such as 3 to about 50 mg per kilogram
body weight of the recipient per day, preferably in the range of 6
to 90 mg/kg/day, most preferably in the range of 15 to 60
mg/kg/day.
[0083] The quinazoline compounds are conveniently administered in
unit dosage form; for example, containing 5 to 1000 mg,
conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of
active ingredient per unit dosage form.
[0084] Ideally, the quinazoline compounds should be administered to
achieve peak plasma concentrations of from about 0.5 to about 75
.mu.M, preferably, about 1 to 50 .mu.M, most preferably, about 2 to
about 30 .mu.M. This may be achieved, for example, by the
intravenous injection of a 0.05 to 5% solution of the quinazoline
compounds, optionally in saline, or orally administered as a bolus
containing about 1-100 mg of the quinazoline compounds. Desirable
blood levels may be maintained by continuous infusion to provide
about 0.01-5.0 mg/kg/hr or by intermittent infusions containing
about 0.4-15 mg/kg of the quinazoline compounds.
[0085] The quinazoline compounds may conveniently be presented in a
single dose or as divided doses administered at appropriate
intervals, for example, as two, three, four or more sub-doses per
day. The sub-dose itself may be further divided, e.g., into a
number of discrete loosely spaced administrations; such as multiple
inhalations from an insulator or by application of a plurality of
drops into the eye.
[0086] Targeting Quinazolines to Cells
[0087] In a preferred embodiment, the quinazoline compound is
targeted to cells where treatment is desired, for example, to
leukemia cells, to breast cells, or to other tumor cells. The
compound is targeted to the desired cell by conjugation to a
targeting moiety that specifically binds the desired cell, thereby
directing administration of a conjugated molecule. Useful targeting
moieties are ligands which specifically bind cell antigens or cell
surface ligands, for example, antibodies against the B cell
antigen, CD19 (such as B43) and the like.
[0088] To form the conjugates of the invention, targeting moieties
are covalently bonded to sites on the quinazoline compound. The
targeting moiety, which is often a polypeptide molecule, is bound
to compounds of the invention at reactive sites, including
NH.sub.2, SH, CHO, COOH, and the like. Specific linking agents are
used to join the compounds. Preferred linking agents are chosen
according to the reactive site to which the targeting moiety is to
be attached.
[0089] Methods for selecting an appropriate linking agent and
reactive site for attachment of the targeting moiety to the
compound of the invention are known, and are described, for
example, in Hermanson, et al., Bioconjugate Techniques, Academic
Press, 1996; Hermanson, et al., Immobilized Affinity Ligand
Techniques, Academic Press, 1992; and Pierce Catalog and Handbook,
1996, pp. T155-T201.
[0090] Administration of Quinazolines
[0091] According to the invention, quinazoline compounds may be
administered prophylactically, i.e., prior to onset the
pathological condition, or the quinazoline compounds may be
administered after onset of the reaction, or at both times.
EXAMPLES
[0092] The invention may be further clarified by reference to the
following Examples, which serve to exemplify some of the preferred
embodiments, and not to limit the invention in any way.
Example 1
Synthesis of Quinazoline Derivatives
[0093] All chemicals were purchased from the Aldrich Chemical
Company, Milwaukee, Wis., and were used directly for synthesis.
Anhydrous solvents such as acetonitrile, methanol, ethanol, ethyl
acetate, tetrahydrofuran, chloroform, and methylene chloride were
obtained from Aldrich as sure seal bottles under nitrogen and were
transferred to reaction vessels by cannulation. All reactions were
carried out under a nitrogen atmosphere.
[0094] The key starting material,
4-chloro-6,7-dimethoxyquinazoline, was prepared according to
published procedures (Nomoto, et al., 1990, Chem. Pharm. Bull.,
38:1591-1595; Thomas, C. L., 1970, IN:Catalytic Processes and
Proven Catalysts, Academic Press, New York, N.Y.) as outlined below
in Scheme 1. Specifically, 4,5-dimethoxy-2-nitrobenzoic acid
(compound 1) was treated with thionyl chloride to form acid
chloride, followed by reacting with ammonia to yield
4,5-dimethoxy-2-nitrobenzamide (compound 2). Compound 2 was reduced
with sodium borohydride in the presence of catalytic amounts of
copper sulphate to give 4,5-dimethoxy-2-aminobenzami- de (compound
3), which was directly refluxed with formic acid to yield
6,7-dimethoxyquinazoline-4(3H)-one (compound 4). Compound 4 was
refluxed with phosphorus oxytrichloride to give
4-chloro-6,7-dimethoxyquinazoline (compound 5) in good yield. 4
[0095] Substituted quinazoline derivatives were prepared by the
condensation of 4-chloro-6,7-dimethoxyquinazoline with substituted
anilines as outlined below in Scheme 2: 5
[0096] Specifically, a mixture of 4-chloro-6,7-dimethoxyquinazoline
(448 mg, 2 mmols) and the substituted aniline (2.5 mmols) in EtOH
(20 ml) was heated to reflux. After refluxing for 4-24 hours, an
excess amount of Et.sub.3N was added, and the solvent was
concentrated to give the crude product which was recrystalized from
DMF.
[0097] As discussed above, the novel hydroxy-substituted
quinazoline derivatives of the invention were created by reacting
the appropriate substituted anilines with the key starting
material, 4-chloro-6,7-dimethoxyquinazoline.
[0098] Physical Characteristics:
[0099] Melting points are uncorrected. .sup.1H NMR spectra were
recorded using a Varian Mercury 300 spectrometer in DMSO-d.sub.6 or
CDCl.sub.3. Chemical shifts are reported in parts per million (ppm)
with tetramethylsilane (TMS) as an internal standard at zero ppm.
Coupling constants (3) are given in hertz and the abbreviations s,
d, t, q, and m refer to singlet, doublet, triplet, quartet and
multiplet, respectively. Infrared spectra were recorded on a
Nicolet PROTEGE 460-IR spectrometer. Mass spectroscopy data were
recorded on a FINNIGAN MAT 95, VG 7070E-HF G.C. system with an HP
5973 Mass Selection Detector. UV spectra were recorded on BECKMAN
DU 7400 and using MEOH as the solvent. TLC was performed on a
precoated silica gel plate (Silica Gel KGF; Whitman Inc). Silica
gel (200-400 mesh, Whitman Inc.) was used for all column
chromatography separations. All chemicals were reagent grade and
were purchased from Aldrich Chemical Company (Milwaukee, Wis.) or
Sigma Chemical Company (St. Louis, Mo.).
Example 2
Bromine Substituted Quinazoline Compounds
[0100] Bromine substituted quinazoline derivatives were synthesized
and characterized as discussed above in Example 1. The structures
and physical data are shown below:
1 Bromine Substituted Quinazoline Compounds No Name Structure
Formula MW 1 P-79 6 C.sub.16H.sub.14BrN.sub.3O.sub.2 360 2 P-88 7
C.sub.17H.sub.14BrN.sub.3O.sub.4 404 3 P-97 8
C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.3 455 4 P-111 9
C.sub.17H.sub.16BrN.sub.3O.sub.2 374 5 P-112 10
C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.2 439 6 P-154 11
C.sub.16H.sub.14BrN.sub.3O.sub.3 376 7 P-160 12
C.sub.23H.sub.18BrN.sub.3O.sub.2 448 8 P-164 13
C.sub.17H.sub.13BrN.sub.2O.sub.3 373 9 P-190 14
C.sub.17H.sub.16BrN.sub.3O.sub.3 389 10 P-210 15
C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2 453 11 P-211 16
C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2 453 12 P-212 17
C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2 453 13 P-214 18
C.sub.16H.sub.13BrFN.sub.3O.sub.2 378 14 P-222 19
C.sub.16H.sub.12Br.sub.3N.sub.3O.sub.2 518 15 P-234 20
C.sub.17H.sub.17N.sub.3O.sub.2 295 16 P-241 21
C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2 453 17 P-258 22
C.sub.16H.sub.15N.sub.3O.sub.2 281 18 P-260 23
C.sub.16H.sub.14BrN.sub.3O.sub.2 360 19 P-261 24
C.sub.16H.sub.14BrN.sub.3O.sub.2 360 20 P-262 25
C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.2 439 21 P-263 26
C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.2 439
4-(3'-Bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P79) Yield
84.17%; m.p. 246.0-249.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 10.42(br, s, 1H, NH), 8.68(s, 1H, 2-H), 8.07-7.36(m, 5H,
5,2',4',5',6'-H), 7.24(s, 1H, 8H), 3.98(s, 3H, --OCH.sub.3),
3.73(s, 3H, --OCH.sub.3); IR(KBr).nu..sub.max: 3409, 2836, 1632,
1512, 1443, 1243, 1068 cm.sup.-1; GC/MS m/z 361(M.sup.+ + 1, 61.8),
360(M.sup.+, 100.0), 359(M.sup.+ - 1, 63.5), 344(11.3), 222(10.9),
140(13.7). Anal. (C.sub.16H.sub.14BrN.sub.3O.sub.2HCl) C, H, N.
4-(4'-Bromo-2'-caboxylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P88) Yield 92.82%; m.p. > 300.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6 + CF.sub.3CO.sub.2H): .delta. 9.95(d, 1H), 8.74(d,
1H, Ar--H), 8.30, 8.28(2d, 2H), 7.95(d, 1H), 7.83(s, 1H), 4.21(s,
3H, --OCH.sub.3), 4.15(s, 3H, --OCH.sub.3). UV(MeOH): 205, 229.0
nm. IR(KBr).nu..sub.max: 3444(br), 2737, 1592, 1504, 1443, 1273,
1070 cm.sup.-1. GC/MS m/z 388(M.sup.+ + 1 --OH, 18.08), 387(M.sup.+
--OH, 100.00), 386(M.sup.+ - 1 --OH, 30.84), 385(97.52), 299(4.78).
Anal. (C.sub.16H.sub.14BrN.sub.3O.sub.2HCl) C, H, N.
4-(3',5'-Dibromo-4'-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P97). Yield 72.80%; m.p. > 300.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 9.71(s, 1H, --NH), 9.39(s, 1H, --OH),
8.48(s, 1H, 2-H), 8.07(s, 2H, 2',6'-H), 7.76(s, 1H, 5-H), 7.17(s,
1H, 8-H), 3.94(s, 3H, --OCH.sub.3), 3.91(s, 3H, --OCH.sub.3).
UV(MeOH): 208.0, 210.0, 245.0, 320.0 nm; IR(KBr).nu..sub.max:
3504(br), 3419, 2868, 1627, 1512, 1425, 1250, 1155 cm.sup.-1; GC/MS
m/z 456(M.sup.+ + 1, 54.40), 455(M.sup.+, 100.00), 454(M.sup.+ - 1,
78.01), 439(M.sup.+ --OH, 7.96), 376(M.sup.+ + 1 --Br, 9.76),
375(M.sup.+ --Br, 10.91), 360(5.23). Anal.
(C.sub.16H.sub.13Br.sub.2N.sub- .3O.sub.3) C, H, N.
4-(3'-Bromo-4'-methylphenyl)-amino-6,7-dimethox- yquinazoline
(HI-P111): Yield 82.22%; m.p. 225.0-228.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 10.23(s, 1H, --NH), 8.62(s, 1H, 2-H),
8.06(d, 1H, J.sub.2',6'=2.1 Hz, 2'-H), 7.89(s, 1H, 5-H), 7.71(dd,
1H, J.sub.5',6'=8.7 Hz, J.sub.2',6'=2.1 Hz, 6'-H), 7.37(d, 1H,
J.sub.5',6'=8.7 Hz, 5'-H), 7.21(s, 1H, 8-H), 3.96(s, 3H,
--OCH.sub.3), 3.93(s, 3H, --OCH.sub.3). UV(MeOH): 204.0, 228.0,
255.0, 320.0 nm. IR(KBr).nu..sub.max: 3431, 3248, 2835, 1633, 1517,
1441, 1281, 1155 cm.sup.-1. GC/MS m/z 375(M.sup.+ + 1, 76.76),
374(M.sup.+, 100.00), 373(M.sup.+ - 1, 76.91), 358(M.sup.+ + 1
--OH, 11.15), 357(1.42), 356(6.31). Anal.
(C.sub.17H.sub.16BrN.sub.3O.sub.2.HCl) C, H, N.
4-(2',5'-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P112):
Yield 70.05%; m.p. > 300.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 11.51(s, 1H, --NH), 8.76(s, 1H, 2-H), 8.21(s, 1H, 5-H),
7.81(d, 1H, J.sub.4',6'=2.4 Hz, 6'-H), 7.75(d, 1H, J.sub.3',4'=8.7
Hz, 3'-H), 7.55(dd, 1H, J.sub.4',6'=2.4 Hz, J.sub.3',4'=8.7 Hz,
4'-H), 7.33(s, 1H, 8-H), 3.98(s, 3H, --OCH.sub.3), 3.97(s, 3H,
--OCH.sub.3). UV(MeOH): 208.0, 238.0, 330.0 nm.
IR(KBr).nu..sub.max: 3444, 2836, 1628, 1510, 1431, 1277, 1070
cm.sup.-1. GC/MS m/z 440(M.sup.+ + 1, 10.12), 439(M.sup.+, 7.0),
438(M.sup.+ - 1, 3.63), 360(M.sup.+ + 1 --Br, 99.42), 359(M.sup.+
--Br, 20.45), 358(M.sup.+ - 1 --Br, 100.00), 343(20.80), 299(8.62).
Anal. (C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.2.HCl) C, H, N.
4-[(3'-Bromo-9'-fluorenone)-2']-amino-6,7-dimethoxyquinazoline
(HI-P119): Yield 75.23%; m.p. 255.0-257.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 8.77(s, 1H, --NH), 8.33(s, 1H, 2-H).
7.89(s, 1H, 5-H), 7.40(s, 1H, 8-H), 7.74-7.26(m, 6H, Ar--H),
4.12(s, 3H, --OCH.sub.3), 4.11(s, 3H, --OCH.sub.3). UV(MeOH): 205,
229.0, 251.0, 320.0 nm. IR(KBr).nu..sub.max: 3444, 2836, 1628,
1510, 1431, 1277, 1070 cm.sup.-1. GC/MS m/z 464(M.sup.+ + 2,
40.81), 463(M.sup.+ + 1, 7.56), 462(M.sup.+, 27.26), 384(M.sup.+ +
2 --Br, 69.56), 383(M.sup.+ + 1 --Br, 35.50), 382(M.sup.+ --Br,
100.00), 352(10.85), 206(26.73), 191(11.31). Anal.
(C.sub.23H.sub.16BrN.sub- .3O.sub.3HCl) C, H, N.
4-(2',3',5',6'-Tetrafluoro-4'-bromolphenyl)a-
mino-6,7-dime-thoxyquinazoline (HI-P144: Yield 78.24%; m.p.
180.0-182.0.degree. C. .sup.1H NMR(DMSO-d.sub.6): .delta. 7.78(s,
1H, 2-H), 7.53(s, 1H, 5-H), 6.79(s, 1H, 8-H), 3.81(s, 3H,
--OCH.sub.3), 3.3.79(s, 3H, --OCH.sub.3). Anal(C.sub.16H.sub.10BrF-
.sub.4N.sub.3O.sub.2.HCl) C, H, N.
4-(3'-Bromo-4'-hydroxylphenyl)-a- mino-6,7-dimethoxyquinazoline
(HI-P154): Yield 89.90%; m.p. 233.0-233.5.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 10.08(s, 1H, --NH), 9.38(s, 1H, --OH),
8.40(s, 1H, 2-H), 7.89(d, 1H, J.sub.2',6'=2.7 Hz, 2'-H), 7.75(s,
1H, 5-H), 7.55(dd, 1H, J.sub.5',6'=9.0 Hz, J.sub.2',6'=2.7 Hz,
6'-H), 7.14(s, 1H, 8-H), 6.97(d, 1H, J.sub.5',6'=9.0 Hz, 5'-H),
3.92(s, 3H, --OCH.sub.3), 3.90(s, 3H, --OCH.sub.3). UV(MeOH):
203.0, 222.0, 250.0, 335.0 nm. IR(KBr).nu..sub.max: 3431(br), 2841,
1624, 1498, 1423, 1244 cm.sup.-1. GC/MS m/z 378(M.sup.+ + 2,
90.68), 377(M.sup.+ + 1, 37.49), 376(M.sup.+, 100.00), 360(M.sup.+,
3.63), 298(18.86), 282(6.65). Anal.
(C.sub.16H.sub.14BrN.sub.3O.sub.3.HCl) C, H, N.
4-[(7'-Bromofluorene)-2']-amino-6,7-dimethoxyquinazoline (HI-P160):
Yield 73.21%; m.p. 254.0-256.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 9.69(br, s, 1H, --NH), 8.52(s, 1H, 2-H),
8.12-7.20(m, 9H, 5, 8, 1', 3', 4', 5', 6', 8', 9'-H), 3.99(s, 3H,
--OCH.sub.3), 3.94(s, 3H, --OCH.sub.3). UV(MeOH): 208.0, 223.0,
348.0 nm. IR(KBr).nu..sub.max: 3421, 2820, 1624, 1516, 1431, 1294,
1223 cm.sup.-1. GC/MS m/z 450(M.sup.+ + 2, 100), 449(M.sup.+ + 1,
35), 448(M.sup.+, 95), 311(25). Anal.
(C.sub.23H.sub.18BrN.sub.3O.sub.2- .HCl) C, H, N.
4-(3'-Bromobenzoyl)-6,7-dimethoxyquinazoline (HI-P164). Yield
81.20%, m.p. 258.0-263.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 9.25(s, 1H, 2-H), 8.14(s, 1H, 5-H), 7.92-7.43(m, 4H,
2',4',5',6'-H), 7.40(s, 1H, 8-H), 4.11(s, 3H, --OCH.sub.3), 4.00(s,
3H, --OCH.sub.3). UV(MeOH): 203.0, 220.0, 238.0 nm.
IR(KBr).nu..sub.max: 3432, 1664, 1504, 1431, 1230 cm.sup.-1. GC/MS
m/z 374(M.sup.+ + 1, 48.96), 373(M.sup.+, 34.93), 372(M.sup.+ - 1,
47.67), 357(58.74), 343(100.00), 293(M.sup.+ --Br, 31.48),
189(26.27). Anal. (C.sub.17H.sub.13BrN.sub.2O.sub.3) C, H, Br, N.
4-(4'-Bromo-6'-hydroxymethylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P190): Yield 73.08%; m.p. 222.0-223.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.30(s, 1H, --OH), 8.22(s, 1H, --NH),
7.77-7.23(m, 5H, 5,8,2',3',5'-H), 4.49(s, 2H, PhCH.sub.2--H),
4.01(s, 3H, --OCH.sub.3), 3.90(s, 3H, --OCH.sub.3). UV(MeOH):
207.0, 250.0, 332.0 nm. IR(KBr).nu..sub.max: 3446, 2829, 2752,
1652, 1560, 1471, 1365, 1280 cm.sup.-1. GC/MS m/z 391(M.sup.+ + 1,
29.33), 389(M.sup.+, 29.82), 360(M.sup.+ --CH.sub.2OH, 50.76),
358(52.39), 311(18.33), 280(43.20), 206(62.80), 191(100.00). Anal.
(C.sub.17H.sub.16BrN.su- b.3O.sub.3.HCl) C, H, N.
4-(2',3'-Dibromo-4'-methylphenyl)-amino-6,- 7-dimethoxyquinazoline
(HI-P210): Yield 81.24%, mp 233.0-236.0.degree. C., .sup.1H
NMR(DMSO-d.sub.6): .delta. 8.55(s, 1H, --NH), 8.08(s, 1H, 2-H),
7.33-7.17(m, 4H, 5,8,5',6'-H), 3.89(s, 6H, --OCH.sub.3), 2.35(s,
3H, --CH.sub.3). UV(MeOH): 207.0, 232.0, 247.0, 330.0 nm.
IR(KBr).nu..sub.max: 3448, 2840, 1629, 1580, 1525, 1420, 1281
cm.sup.-1. GC/MS m/z 454(M.sup.+ + 1, 4.45) , 453(M.sup.+, 11.31),
452(M.sup.+ - 1, 4.45), 375(20.36), 374(97.59), 373( 23.55),
372(100.00), 358(19.61), 356(18.43). Anal.
(C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2.HCl) C, H, N.
4-(2',5'-Dibromo-4'-methylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P211): Yield 83.50%; m.p. 282.0-284.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.30(s, 1H, --NH), 8.58(s, 1H, 2-H),
8.00(s, 1H, 5-H), 7.65(s, 1H, 6'-H), 7.60(s, 1H, 3'-H), 7.13(s, 1H,
8-H), 3.79(s, 3H, --OCH.sub.3), 3.78(s, 3H, --OCH.sub.3), 2.29(s,
3H, --CH.sub.3). UV(MeOH): 207.0, 239.0, 330.0 nm.
IR(KBr).nu..sub.max: 3442, 2620, 1631, 1580, 1514, 1380, 1280
cm.sup.-1. GC/MS m/z 454(M.sup.+ + 1, 5.86), 453(M.sup.+, 16.16),
452(M.sup.+ - 1, 5.35), 374(92.12), 373(23.66), 372(100.00),
358(17.68), 356(17.35). Anal.
(C.sub.17H.sub.15Br.sub.2N.sub.3O.sub.2.HCl) C, H, N.
4-(3',5'-Dibromo-4'-methylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P212): Yield 83.47%; m.p. 275.0-279.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.30(s, 1H, --NH), 8.58(s, 1H, 2-H),
8.35(s, 1H, 5-H), 7.24(s, 2H, 2',6'-H), 7.13(s, 1H, 8-H), 3.91(s,
3H, --OCH.sub.3), 3.88(s, 3H, --OCH.sub.3), 2.31(s, 3H,
--CH.sub.3). UV(MeOH): 237.0, 307.0, 319.0 nm. IR(KBr).nu..sub.max:
3471, 3434, 2640, 1633, 1580, 1504, 1420, 1281 cm.sup.-1. GC/MS m/z
454(M.sup.+ + 1, 5.34), 453(M.sup.+, 16.05), 452(M.sup.+ - 1,
5.87), 374(99.02), 373(26.20), 372(100.00), 358(20.39), 356(19.98),
32(8.29), 314(8.49), 206(19.02). Anal. (C.sub.17H.sub.15Br.sub.2N.-
sub.3O.sub.2HCl) C, H, N.
4-(2'-Fluoro-4'-bromophenyl)-amino-6,7-di- methoxyquinazoline
(HI-P214): Yield 77.21%; m.p. 243.0-245.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 8.57(s, 1H, 2-H), 7.91(s, 1H, 5-H),
7.57(d, 1H, 3'-H), 7.34(m, 2H, 5',6'-H), 7.07(s, 1H, 8-H), 3.78(s,
3H, --OCH.sub.3), 3.77(s, 3H, --OCH.sub.3). UV(MeOH): 204.0, 215.0,
250.0, 330.0 nm. IR(KBr).nu..sub.max: 3431, 2629, 1633, 1580, 1511,
1420, 1278 cm.sup.-1. GC/MS m/z 379(M.sup.+ + 1, 34.39),
378(M.sup.+, 21.33), 377(M.sup.+ - 1, 39.08), 360(62.05),
359(31.58), 358(62.57), 357(19.81), 299(19.31), 298(100.00),
282(17.88), 240(28.76). Anal.
(C.sub.16H.sub.13BrFN.sub.3O.sub.2HCl) C, H, N.
4-(2',4',6'-Tribromophenyl)amino-6,7-dimethoxyquinazoline
(HI-P222): Yield 54.86%; m.p. 250.0-255.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 8.00(s, 1H, 2-H), 7.89(s, 2H, 3', 5'-H),
7.74(s, 1H, 5-H), 7.01(s, 1H, 8-H), 3.87(s, 3H, --OCH.sub.3),
3.86(s, 3H, --OCH.sub.3). UV(MeOH): 209.0, 236.0, 333.0 nm.
IR(KBr).nu..sub.max: 3417, 2838, 1625, 1514, 1429, 1276, 1073
cm.sup.-1. GC/MS m/z 519(M.sup.+ + 1, 18.12), 518(M.sup.+, 17.30),
517(M.sup.+ - 1, 16.63), 439(M.sup.+ + 1-Br, 99.42), 438(M.sup.+ -
Br, 95.45), 437(M.sup.+ - 1 --Br, 100.00), 359(20.80), 358(18.62),
357(19.32), 281(88.98), 207(15.42). Anal.
(C.sub.16H.sub.12Br.sub.3N.sub.3O.sub.2HCl) C, H, N.
4-(2',6'-Dibromo-4'-methylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P241): Yield 79.47%, m.p. 235.0-237.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 9.77(s, 1H, --HN), 8.20(s, 1H, 2-H),
7.87(s, 1H, 8-H), 7.61(s, 2H, 3',5'-H), 7.15(s, 1H, 5-H), 3.93(s,
6H, --OCH.sub.3). UV(MeOH): 208.0, 245.0, 318.0, 339.0 nm.
IR(KBr).nu..sub.max: 3241, 2839, 2783, 1635, 1580, 1514, 1420,
1360, 1281 cm.sup.-1. GC/MS m/z 454(M.sup.+ + 1, 7.86),
453(M.sup.+, 56.16), 452(M.sup.+ - 1, 15.30), 374(95.12),
373(18.66), 372(100.00), 358(29.64), 356(19.36). Anal.
(C.sub.17H.sub.15Br.sub.2N.sub- .3O.sub.2HCl) C, H, N.
4-(4'-Bromophenyl)-amino-6,7-dimethoxyquinaz- oline (HI-P260):
Yield 75.28%. m.p. 270.0-272.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.30(s, 1H, --NH), 8.85(s, 1H, 2-H),
8.27(s, 1H, 5-H), 7.70(s, 4H, 2',3',5',6'-H), 7.32(s, 1H, 8H),
4.02(s, 3H, --OCH.sub.3). 4.00(s, 3H, --OCH.sub.3). UV(MeOH):
204.0, 218.0, 252.0, 335.0 nm. IR(KBr).nu..sub.max: 3431, 3034,
2636, 1635, 1589, 1514, 1435, 1284 cm.sup.-1. GC/MS m/z 361(M.sup.+
+ 1, 74.00), 360(M.sup.+, 100.00), 359(M.sup.+ - 1, 72.00),
358(M.sup.+ - 2, 95.00), 329(3.20), 301(13.0), 281(21.0),
207(38.0). Anal. (C.sub.16H.sub.14BrN.sub.3O.sub.2.HCl) C, H, N.
4-(2'-Bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P261): Yield
71.94%; m.p. 241.0-243.0.degree. C. .sup.1H NMR(DMSO-d.sub.6)
.delta. 11.67(d, 1H, --NH), 8.79(s, 1H, 2-H), 8.32(s, 1H, 5-H),
7.86-7.38(m, 4H, 3',4',5',6'-H), 7.40(s, 1H, 8H), 4.01(s, 6H,
--OCH.sub.3). UV(MeOH): 204.0, 226.0, 248.0, 330.0 nm.
IR(KBr).nu..sub.max : 3454, 3032, 2638, 1630, 1589, 1514, 1430,
1281 cm.sup.-1. GC/MS m/z 361(M.sup.+ + 1, 7.00), 360(M.sup.+,
5.00), 359(M.sup.+ - 1, 6.00), 358(M.sup.+ - 2, 5.00), 301(13.0),
281(21.0), 280(100.00), 207(25.00). Anal
(C.sub.16H.sub.14BrN.sub.3O.sub.2.HC- l) C, H, N.
4-(2',6'-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P262):
Yield 69.45%, mp 243.0-246.0.degree. C., .sup.1H NMR(DMSO-d.sub.6):
.delta. 11.91(d, 1H, --NH), 8.80(s, 1H, 2-H), 8.43(s, 1H, 5-H),
7.86(d, 2H, J=8.4 Hz, 3',5'-H), 7.49(s, 1H, 8H), 7.35(t, 1H, J=8.4
Hz, 4'-H), 4.02(s, 3H, --OCH.sub.3), 4.01(s, 3H, --OCH.sub.3).
UV(MeOH): 208.0, 227.0, 245.0, 330.0 nm. IR(KBr).nu..sub.max: 3454,
3032, 2638, 1630, 1589, 1514, 1430, 1281 cm.sup.-1.
4-(2',4'-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P263):
Yield 70.62%; m.p. 257.0-262.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.91(d, 1H, --NH), 8.79(s, 1H, 2-H),
8.21(s, 1H, 5-H), 8.12-7.51(m, 3H, 3',5',6'-H), 7.35(s, 1H, 8-H),
4.01(s, 3H, --OCH.sub.3), 3.99(s, 3H, --OCH.sub.3). UV(MeOH):
208.0, 210.0, 248.0, 330.0 nm. IR(KBr).nu..sub.max: 3458, 3028,
2641, 1633, 1594, 1511, 1435, 1277 cm.sup.-1.
Example 3
[0101]
2 Chlorine Substituted Quinazoline Compounds Chlorine substituted
quinazoline derivatives were synthesized and characterized as
discussed above in Example 1. The structures and physical data are
shown below: No Name Structure Formula MW 1 P-87 27
C.sub.16H.sub.14ClN.sub.3O.sub.2 316 2 P-93 28
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 3 P-189 29
C.sub.16H.sub.13Cl.sub.2N.sub.3O.sub.3 365 4 P-197 30
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 5 P-268 31
C.sub.16H.sub.14ClN.sub.3O.sub.2 316 6 P-269 32
C.sub.16H.sub.14ClN.sub.3O.sub.2 316 7 P-278 33
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 8 P-415 34
C.sub.20H.sub.16ClN.sub.3O.sub.2 365
4-(3'-Chlorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P87). Yield
76.98%; m.p. 242.0-245.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 10.47(br, s, 1H, NH), 8.69(s, 1H, 2-H), 8.06(s, 1H, 5-H),
7.95-7.23(m, 4H, 2',4',5',6'-H), 7.24(s, 1H, 8-H), 3.98(s, eH,
--OCH.sub.3), 3.35(s, 3H, 0OCH.sub.3). UV(MeOH): 228.0, 251.0,
332.0 nm. IR(KBr).nu..sub.max: 3406, 2839, 1632, 1516, 1443, 1278,
1068 cm.sup.-1. GC/MS m/z 316(M.sup.+ - 1, 68.34), 314(M.sup.+ - 2,
100.00, 344(11.34), 222(4.35), 140(9.86). Found: C, 54.62; H, 4.68;
N, 11.93; Cl, 19.23. C.sub.16H.sub.14ClN.sub.3O.sub.2.HCl requires:
C, 54.70; H, 4.28; N, 11.96; Cl, 19.96%. 4-(c'-Chloroo-6'-hydroxyp-
henyl)amino-6,7-dimethoxyquinazoline (HI-P93) Yield 83.08%; m.p.
295.0.degree. C.(dec). .sup.1H NMR9DMSO-d.sub.6: .delta. 10.14(s,
1H, --OH), 8.37(s, 1H, 2-H), 7.78(s, 1H, 5H), 7.57(d, 1H,
J.sub.2',4'=2.4 Hz, 2'-H), ), 7.16(s, 1H, 8-H), 7.07(dd, 1H,
J.sub.2',4'=2.4 Hz, J.sub.4',5'=8.7 Hz, 4'-H), 6.92(d, 1H,
J.sub.4',5'=8.7 Hz, 5'-H), 3.93(s, 3H, --OCH.sub.3). 3.92(s, 3H,
--OCH.sub.3). UV(MeOH): 205, 229.0, 251.0, 320.0 nm.
IR(KBr).nu..sub.max: 3500(br), 3430, 2835, 1622, 1512, 1432, 1259
cm.sup.-1. GC/MS m/z 333(M.sup.+ + 2, 13.41), 332(M.sup.+ + 1,
9.73, 331(M.sup.+, 39.47), 314(M.sup.+ --OH, 100.00), 298(7.64).
Found: C, 52.25; H, 4.07; N, 11.39.
C.sub.16H.sub.14ClN.sub.3O.sub.3.HCl requires: C, 52.32; H, 4.09;
N, 11.44%. 4-(4'-Hydroxyl-3',5'-dichlorophenyl)ami-
no-6,7-dimethoxyquinazoline (HI-P189) Yield 79.45%; m.p.
293.0-295.0.degree. C. .sup.1H NMR-DMSO-d.sub.6): .delta. 11.32(s,
1H, --NH), 10.34(s, 1H, --OH), 8.87(s, 1H, 2-H), 8.29(s, 1H, 5-H),
7.90(s, 2H, 2',6'-H), 7.32(s, 1H, 8-H), 4.01(s, 3H, --OCH.sub.3),
3.99(s, 3H, --OCH.sub.3). UV(MeOH): 213.0, 232.0, 250.0, 335.0 nm.
IR(KBr).nu..sub.max: 3479, 2564, 1641, 1579, 1429, 1282, 1147
cm.sup.-1. GC/MS m/z 367(M.sup.+ = 2, 66.57), 366(M.sup.+=1,
75.91), 365(M.sup.+, 100.00), 364(M.sup.+ - 194.08), 349(M.sup.+
--OH, 11.16). Anal. (C.sub.16H.sub.13Cl.sub.2N.sub.3O.sub.3) C, H,
N. Found: C, 48.93; H, 4.51; N, 10.00.
C.sub.17oH.sub.17Cl.sub.2N.sub.3O.sub- .3.Hcl requires: C, 48.80;
H, 4.31; N, 10.04.%.
4-(3'-Chloro-4'-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P197). Yield 84.14%; m.p. 245.0.degree. C.(dec). .sup.1H
NMR(DMSO-d.sub.6): .delta. 10.00(s, 1H, --NH), 9.37(s, 1H, --OH),
8.41(s, 1H, 2-H), 7.78(s, 1H, 5-H), 7.49(d, 1H, J.sub.2',5'=2.7 Hz,
2'-H), 7.55(dd, 1H, J.sub.5',6'=9.0 Hz, J.sub.2',6'=2.7 Hz, 6'-H),
7.16(s, 1H, 8-H), 6.97(d, 1H, J.sub.5',6'=9.0 Hz, 5'H), 3.93(s, 3H,
--OCH.sub.3), 3.91(s, 3H, --OCH.sub.3). UV(MeOH): 209.0, 224.0,
249.0, 330.0 nm. IR(KBr).nu..sub.max: 3448, 2842, 1623, 1506, 1423,
1241 cm.sup.-1. GC/MX m/z: 341(M.sup.+, 100.00), 326(M.sup.+
--CH.sub.3, 98.50), 310(M.sup.+ --OCH.sub.3, 12.5), 295(9.0.),
189(13.5), 155(13.8). Found: C, 521.35; H, 4.16; Cl, 19.15; N,
11.39. C.sub.16H.sub.14ClN.sub.3O.sub.3.HCl requires: C, 52.32; H,
4.09; Cl, 19.07; N, 11.44%. 4-(2'-Chlorophenyl)-amino-6,7-dimethox-
yquinazoline (HI-P268) Yield 87.28%; m.p. 247.0-279.5.degree. C.
.sup.1H NMR(DMSO-d.sub.6): .delta. 11.71(s, 1H, --NH), 8.78(s, 1H,
2-H), 8.33(s, 1H, 5-H), 7.67(s, 1H, 8H), 7.68-7.42(m, 4H,
3',4,5,6'-H), 4.00(s, 3H, --OCH.sub.3), 3.99(s, 3H, --OCH.sub.3).
UV(MeOH): 213.0, 234.0, 251.0, 331.0 nm. IR(KBr).nu..sub.max: 3479,
2566, 1643, 1577, 1429, 1282, 1147 cm.sup.-1. GC/MX m/z 317(M.sup.+
+ 1, 6.60), 316(M.sup.+, 6.60), 315(M.sup.+ - 1, 18.52),
314(M.sup.+ - 2, 11.11), 281(21.22), 280(M.sup.+ --Cl, 100.00),
264(29.62). Found: C, 54.51; H, 4.41; N, 11.81.
C.sub.16H.sub.14ClN.sub.3O.sub.2.HCl requires: C, 54.45; H, 4.26;
N, 11.93%. 4-(4'-Chlorophenyl)-amino-6,7-dimethoxyquinazoline
(HI-P269) Yield 94.94%. m.p. 248.0-250.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.62(s, 1H, --NH), 8.85(s, 1H, 2-H),
8.42(s, 1H, 5-H), 7.88(d, 2H, J=8.7 Hz, 3', 5', --H), 7.54(d, 2H,
J=8.7 Hz, 2', 6', --H), 7.38(s, 1H, 8-H0, 4.02(s, 3H, --OCH.sub.3),
3.99(s, 3H, --OCH.sub.3). UV(MeOH): 215.0, 230.0, 253.0, mn.
IR(KBr).nu..sub.max: 3477, 2563, 1640, 1578 cm.sup.-1. GC/MX m/z
317(M.sup.+ + 1, 18.18), 316(M.sup.+, 29.55), 315(M.sup.+ - 1,
48.85), 314(M.sup.+ - 2, 61.36), 281(32., 95), 207(100.00). Found:
C, 54.65; H, 4.38; N, 11.92. C.sub.16H.sub.14ClN.sub.3O.sub.2.HCl
requires: C, 54.55; H, 4.26; N, 11.93%. 4-(4'-Hydroxyl-2'-chloroph-
enyl)-amino-6,7-dimethoxy-quinazoline (HI-P278) Yield 81.44%; m.p.
245.0-247.0.degree. C. .sup.1H NMR(DMSO-d.sub.6): .delta. 11.39(s,
1H, --NH), 10.30(s, 1H, --OH), 8.75(s, 1H, 2-H), 8.24(s, 1H, 5-H),
7.38-6.85(m, 3H, 3',5',6'-H), 7.37(s, 1H, 8H), 3.98(s, 3H,
--OCH.sub.3), 3.96(s, 3H, --OCH.sub.3). UV(MeOH): 222.0, 234.0,
239.0, 245.0, 254.0, 348.0 nm. IR(KBr).nu..sub.max: 3448, 3242,
3144, 3025, 2917, 2834, 1638, 1591, 1514, 1437, 1365, 1277, 1209
cm.sup.-1. GC/MS m/z: 332(M.sup.+ + 1, 5.00), 331(M.sup.+, 17.00),
330(M.sup.+ - 1, 5.00), 297(17.00), 296(100.00), 281(18.00),
280(29.00), 253(9.00). Found: C, 52.17; 11, 4.06; N, 11.32.
C.sub.16H.sub.14ClN.sub.3O.sub.3.HCl requires: C, 52.32; H, 4.01;
N, 11.44%. 4-(4'-Chloronaphthy-1')-amino-6,7-dimethoxyquinazoline
(HI-P415) Yield, 85.07%. m.p. 245.0-248.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.91(s, 1H, --NH), 8.66(s, 1H, 2-H),
8.45(s, 1H, 5-H), 8.30-7.62(m, 6H, 2',3',5',6',7',8'-H), 7.38(s,
1H, 8-H), 4.03(s, 3H, --OCH.sub.3), 4.01(s, 3H, --OCH.sub.3).
UV(MeOH): 211.0, 233.0, 250.0, mn. IR(KBr).nu..sub.max: 3481, 2567,
1645, 1579 cm.sup.-1. Found: C, 59.32; H, 4.27; N, 10.24.
C.sub.20H.sub.16ClN.sub.3O.sub.2.HCl requires: C, 59.70; 11, 4.23;
N, 10.48%.
Example 4
Iodine Substituted Quinazoline Compounds
[0102] Iodine substituted quinazoline derivatives were synthesized
as discussed above in Example 1, and analyzed. The structures and
physical data are shown below:
3 Iodine Substituted Quinazoline Compounds No Name Structure
Formula MW 1 P-270 35 C.sub.16H.sub.14IN.sub.3O.sub.2 407 2 P-271
36 C.sub.16H.sub.14IN.sub.3O.sub.2 407 3 P-300 37
C.sub.16H.sub.14IN.sub.3O.sub.2 407 4 P-294 38
C.sub.16H.sub.13I.sub.2N.sub.3O.sub.3 549 5 P-299 39
C.sub.16H.sub.14IN.sub.3O.sub.3 423
4-(2'-Iodophenyl)-amino-6,7-dimethoxyquinazoline (P-270): Yield
75.37%; m.p. 225.0-230.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 11.74(s, 1H, --NH), 8.79(s, 1H, 2-H), 8.33(s, 1H, 5-H),
8.05-7.13(m, 4H, 3',4,5,6'-H), 7.44(s, 1H, 8H), 4.01(s, 6H,
--OCH.sub.3). UV(MeOH): 219.0, 222.0, 253.0, 342.0 nm.
IR(KBr).nu..sub.max: 3165, 3027, 2827, 1639, 1572, 1501, 1434,
1275, 1070 cm.sup.-1. GC/MS m/z 408(M.sup.+ + 1, 3.47),
407(M.sup.+, 15.28), 406(M.sup.+ - 1, 3.47), 281(33.33),
280(M.sup.+ --I, 100.00), 264(50.00), 207(34.72). Found: C, 43.62;
H, 3.60; N, 9.42. C.sub.16H.sub.14IN.sub.3O.sub.2.HCl requires: C,
43.34; H, 3.38; N, 9.48%.
4-(3'-Iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P271): Yield
79.85%; m.p. 235.0-242.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 11.43(s, 1H, --NH), 8.88(s, 1H, 2-H), 8.33(s, 1H, 5-H),
8.13(s, 1H, 2'-H), 7.80-7.26(m, 3H, 4',5',6'-H), 7.35(s, 1H, 8H),
4.02(s, 3H, --OCH.sub.3), 4.00(s, 3H, --OCH.sub.3). UV(MeOH):
203.0, 210.0, 228.0, 251.0, 331.0 nm. (KBr).nu..sub.max: 3191,
3022, 2940, 2836, 2576, 1629, 1516, 1444, 1276, 1153, 1060
cm.sup.1. GC/MS m/z 406(M.sup.+, 1.52), 405(M.sup.+ - 1, 6.22),
281(35.33), 207(100.00). Found: C, 43.55; H, 3.43; N, 9.32.
C.sub.16H.sub.14IN.sub.3O.sub.2.HCl requires: C, 43.34; H, 3.38; N,
9.48%. 4-(4'-Hydroxy-3,5-diiodophenyl)-amino-6,7-dimethoxy-quinazo-
line (HI-P294: Yield 77.47%; m.p. 259.0-260.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.13(s, 1H, NH), 9.73(s, 1H, --OH),
8.87(s, 1H, 2-H), 8.16(s, 1H, 5-H), 8.09(s, 2H, 2',6'-H), 7.28(s,
1H, 8H), 3.98(s, 6H, --OCH.sub.3). UV(MeOH).lambda..sub.max
(.epsilon.):. 217.0, 227.0, 252.0 nm. IR(KBr).nu..sub.max: 3457,
3201, 2934, 2832, 2566, 1629, 1562, 1521, 1439, 1275, 1075
cm.sup.-1. GC/MS m/z: 422(M.sup.+ --I, 33.53), 405(7.50),
281(86.67), 221(51.80), 207(91.30). Found: C, 32.60; H, 2.50; N,
6.92. C.sub.16H.sub.13I.sub.2N.sub.3O.sub.3.HCl requires: C, 32.82;
H, 2.39; N, 7.18%.
4-(4'-Hydroxy-3'-iodophenyl)-amino-6,7-dimethoxyquinazoline
(HI-P299) Yield 71.59%; m.p. 248.0-250.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.32(d, 1H, NH), 10.62(s, 1H, --OH,
8.79(s, 1H, 2-H), 8.26(s, 1H, 5-H), 7.98-6.98(m, 3H, 2',3',6'-H),
7.32(s, 1H, 8H), 3.98(s, 3H, --OCH.sub.3), 3.97(s, 3H,
--OCH.sub.3). UV(MeOH).lambda..sub.max (.epsilon.): 217.0 , 227.0,
252.0 nm. IR(KBr).nu..sub.max: 3411, 2975, 2730, 2366, 1634, 1573,
1501, 1429, 1229, 1075 cm.sup.-1. GC/MS m/z: 406(M.sup.+ - 1,
3.33), 405(M.sup.+ - 2, 7.50), 281(M.sup.+ - 1 --I, 26.67),
253(11.80), 207(100.00). Found: C, 41.96; H, 3.40; N, 8.98.
C.sub.16H.sub.14IN.sub.3O.sub.3.HCl requires: C, 41.83; H, 3.26; N,
9.15%. 4-(4'-Iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P300):
Yield 85.24%; m.p. 240.0-242.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.51(s, 1H, NH), 8.82(s, 1H, 2-H),
8.37(s, 1H, 5-H), 7.81(d, 2H, J=8.4 Hz, 2',6'-H), 7.55(d, 2H, J=8.4
Hz, 3',5'-H), 7.35(s, 1H, 8H), 4.01(s, 3H, --OCH.sub.3), 3.98(s,
3H, --OCH.sub.3). UV(MeOH): 217.0, 227.0, 252.0 nm.
IR(KBr).nu..sub.max: 3211, 3032, 2832, 2720, 1629, 1573, 1501,
1434, 1235, 1153, 1070 cm.sup.-1. GC/MS m/z 406(M.sup.+ - 1, 3.33),
405(M.sup.+ - 2, 7.50), 281(M.sup.+ - 1 --I, 26.67), 253(11.80),
207(100.00). Found: C, 43.40; H, 3.39; N, 9.36.
C.sub.16H.sub.14IN.sub.3O.sub.2.HCl. requires: C, 43.34; H, 3.38;
N, 9.48%.
Example 5
[0103]
4 OH Group Substituted Quinazoline Compounds OH group substituted
quinazoline derivatives were synthesized and characterized as
discussed above for Example 1. The structures and physical data are
shown below: No Name Structure Formula MW 1 P-93 40
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 2 P-97 41
C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.3 455 3 P-131 42
C.sub.16H.sub.15N.sub.3O.sub.3 297 4 P-132 43
C.sub.16H.sub.15N.sub.3O.sub.3 297 5 P-133 44
C.sub.19H.sub.16N.sub.4O.sub.3 348 6 P-150 45
C.sub.15H.sub.14N.sub.4O.sub.3 298 7 P-154 46
C.sub.16H.sub.14BrN.sub.3O.sub.3 376 8 P-180 47
C.sub.16H.sub.15N.sub.3O.sub.3 297 9 P-182 48
C.sub.17H.sub.15N.sub.3O.sub.5 341 10 P-189 49
C.sub.16H.sub.13Cl.sub.2N.sub.3O.sub.3 365 11 P-190 50
C.sub.17H.sub.16BrN.sub.3O.sub.3 389 12 P-191 51
C.sub.17H.sub.17N.sub.3O.sub.3 311 13 P-192 52
C.sub.16H.sub.15N.sub.3O.sub.4 313 14 P-197 53
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 15 P-215 54
C.sub.14H.sub.13N.sub.5O.sub.4 315 16 P-259 55
C.sub.17H.sub.17N.sub.3O.sub.3 311 17 P-265 56
C.sub.18H.sub.19N.sub.3O.sub.3 325 18 P-266 57
C.sub.18H.sub.19N.sub.3O.sub.3 325 19 P-274 58
C.sub.2OH.sub.17N.sub.3O.sub.3 347 20 P-275 59
C.sub.20H.sub.17N.sub.3O.sub.3 347 21 P-276 60
C.sub.18H.sub.19N.sub.3O.sub.3 325 22 P-277 61
C.sub.28H.sub.23N.sub.3O.sub.3 449 23 P-278 62
C.sub.16H.sub.14ClN.sub.3O.sub.3 331 24 P-289 63
C.sub.18H.sub.19N.sub.3O.sub.5 357 25 P-292 64
C.sub.20H.sub.17N.sub.3O.sub.3 341 26 P-293 65
C.sub.20H.sub.17N.sub.3O.sub.3 341 27 P-294 66
C.sub.16H.sub.13I.sub.2N.sub.3O.sub.3 549 28 P-229 67
C.sub.16H.sub.14IN.sub.3O.sub.3 423 29 P-312 68
C.sub.16H.sub.14N.sub.4O.sub.5 342 30 P-313 69
C.sub.16H.sub.14N.sub.4O.sub.5 342 31 P-315 70
C.sub.16H.sub.14N.sub.4O.sub.5 342 32 P-323 71
C.sub.16H.sub.14N.sub.4O.sub.5 342 4-(3'-Chlooro-6'-hydrox-
ylphenyl)amino-6,7-dimethoxyquinazoline (HI-P93) yield 93.08%; m.p.
295.degree. C.(dec). .sup.-H NMR-DMSO-d.sub.6: .delta. 10.14(s, 1H,
--NH), 9.16(s, 1H, --OH), 8.37(s, 1H, 2-h), 7.78(s, 1H, 5H),
7.57(d, 1H, J.sub.2',2'=2.4 Hz, 2'-H), ), 7.16(s, 1H, 8-H),
7.07(dd, 1H, J.sub.2',4'=2.4 Hz, J.sub.4',5'=8.7 Hz, 4'-H), 6.92(d,
1H, J.sub.4',5'=8.7 Hz, 5'-H), 3.93(s, 3H, --OCH.sub.3), 3.92(s,
3H, --OCH.sub.3. UV(MeOH): 205, 229.0, 251.0, 320.0 nm.
IR(KBr).nu..sub.max: 3500(br), 3430, 2835, 1622, 1512, 1432, 1259
cm.sup.-1. GC/MS m/z 333(M.sup.- = 2, 13.41), 332(M.sup.- = 1,
9.73), 331(M.sup.+, 39.47), 314(M.sup.+ --OH, 100.00), 298(7.64).
Found: C, 52.25; H, 4.07; N, 11.39,
C.sub.16H.sub.14ClN.sub.3O.sub.3.HCl requires: C, 52.32; H, 4.09;
N, 11.44%.
4-(3',5'-Dibromo-4'-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline-(HI-P9-
7). Yield 72.80%; m.p. > 300.000. .sup.1H NMR(DMSO-d.sub.6):
.delta. 9.71(s, 1H, --NH), 9.39(s, 1H, --OH), 8.48(s, 1H, 2-h),
8.07(s, 2H, 2',6'-H), 7.76(s, 1H, 5-H), 7.17(s, 1H, 8-H), 3.94(s,
3H, --OCH.sub.3, 3.91(s, 3H, --OCH.sub.3). UV(MeOH): 208.0, 210.0,
245.0, 320.0 nm; IR(KBr).nu..sub.max: 3504(br), 3419, 2868, 1627,
1512, 1425, 1250, 1155 cm.sup.-1; GC/MS m/z 456(M.sup.1 = 1,
54.40), 455(M.sup.-, 100.00), 454(M.sup.- 1, 78.01), 439(M.sup.-
--OH, 7.96), 376(M.sup.- + 1 --Br, 9.76), 375(M.sup.- Br, 10.91),
360(5.23). Anal. (C.sub.16H.sub.13Br.sub.2N.sub.3O.sub.3) C, H, N.
4-(4'-Hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P131):
yield 84.29%; m.p. 245.0-248.0.degree. C. IR(KBr).nu..sub.max:
3428, 2836, 1635, 1516, 1443, 1234 cm: .sup.1H NMR(DMSO-d.sub.6:
.delta. 11.21(s, 1H, --NH), 9.70(s, 1H, --OH), 8.74(s, 1H, 2-h),
8.22(s, 1H, 5-h), 7.40(d, 2H, J-8.9 Hz, 2',6'-H), 7.29(s, 1H, 8-H),
6.85(d, 2H, J=8.9 Hz, 3',5'-H), 3.98(s, 3H, --OCH.sub.3, 3.97(s,
3H, --OCH.sub.2). GC/MS m/z 298(M.sup.- = 1, 100.00), 297(M.sup.-,
26.6), 296(M.sup.+ - 1, 12.5). Anal.
(C.sub.16H.sub.15N.sub.3O.sub.3HCl) Cl, H, N.
4-(2'-Hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P132):
yield 82.49%; m.p. 255.0-258.0.degree. C. IR(KBr).nu..sub.max:
3500(br), 3425, 2833, 1625, 1512, 1456, 1251, 1068 cm.sup.-1.
.sup.1H NMR(DMSO-d.sub.6): .delta. 9.78(s, 1H, --NH), 9.29(s, 1H,
--OH), 8.33(s, 1H, 2-h), 7.85(s, 1H, 5-H), 7.41-6.83(m, 4H,
3',4',5',6'-H), 7.16(s, 1H, 8-H), 3.93(s, 3H, --OCH.sub.3, 3.92(s,
3H, --OCH.sub.3), 280(M.sup.+ --OH, 10.0). Anal.
(C.sub.16H.sub.15N.sub.3O.sub.3, HCl) C, H, N.
4-[(8'-Hydroxyquiline)-5'-Jamino-6,7-dimethoxyquinazoline (HI-P133)
yield 83.51%; m.p. 238.0-239.0.degree. C. .sub.1H NME(DMSO-d.sub.6:
.delta. 10.12(br, s, 1H, --NH), 8.93-7.09 M, 8H,
2,5,2,2',3',4',6',7'-H), 4.04(s, 3H, --OCH.sub.3), 3.96(s, 3H,
--OCH.sub.3). UV(MeOH): 204.0, 245.0, 332.0 nm.
IR(KBr).nu..sub.max: 3425(br), 2935, 1632, 1510, 1437, 1273
cm.sup.-1. GC/MS m/z 349(M.sup.- = 1, 100.00), 348(m+, 26.56),
307(38.50), 289(21.00).
4-[(3'-Hydroxylpyridine)-2']-amino-6,7-dimethoxyquinaz- oline
(HI-P150) Yield 78.65%; m.p. 185.0-187.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 10.08(br, s, 1H, --NH), 8.52(s, 1H,
2-H), 7.88-7.86(m, 1H, 6'-H), 7.60(s, 1H, 5-H), 7.39-7.35(m, 1H,
4'-H), 7.32(s, 1H, 8-H), 6.63-6.58(m, 1H, 5'-H), 5.96(s, 1H, --OH),
3.97(s, 3H, --OCH.sub.3), 3.94(s, 3H, --OCH.sub.3). UV(MeOH):
204.0, 238.0, 321.0 nm. IR(KBr).nu..sub.max: 3500, 3446, 2960,
1475, 1236, 1375, 1182 cm.sup.-1. GC/MS m/z 299(M.sup.- = 1, 100),
298(M.sup.+, 34), 289(11), 291(9). Found: C, 60.26; H, 4.81; N,
18.68. C.sub.15H.sub.14N.sub.4O.sub.5, requires: C, 60.26; H, 4.81;
N, 18.68%.
4-(3'-Bromo-4'-hydroxylphenyl)-amino-6,7-dimethoxyquinazol- ine
(HI-P154); yield 89.90%; m.p. 233.0-233.5.degree. C. .sup.1H
NMR(DMSO-d.sub.6): 10.08(s, 1h, --NH), 9.38(s, 1H, --OH), 8.40(s,
1H 2-H), 7.89(d, 1H, J.sub.2',6'=2.7 Hz, 2'-H), 7.75(s, 1H, 5-h),
7.55(dd, 1H, J.sub.5',6'=9.0 Hz, J.sub.2',6'=2.7 Hz, 6'-H), 7.14(s,
1H, 8-H), 6.97(d, 1H, J.sub.5',6'=9.0 Hz, 5'-H), 3.92(s, 3H,
--OCH.sub.3), 3.90(s, 3H, --OCH.sub.3). UV(MeOH): 203.0, 222.0,
25.0, 335.0 nm. IR(KBr).nu..sub.max: 3431(br), 2841, 1624, 1498,
1423, 1244 cm.sup.-1. GC/MS m/z 378(M.sup.+ = 2, 90.68),
377(M.sup.+ = 1, 37.49), 376(M.sup.+, 100.00), 360(MK.sup.+, 3.63),
298(28.86), 282(6.65). Anal. (C.sub.16H.sub.14BrN.sub.3O.sub.3,
HCl) C, H, N. 4-(3'-Hydroxyphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P180) Yield 71.55%; m.p. 256.0-258.0.degree. C.
IR(KBr).nu..sub.max: 3394, 2836, 1626, 1508, 1429, 1251 cm.sup.-1.
.sup.1H NMR(DMSO-d.sub.6): 9.41(s, 1H, --NH), 9.36(s, 1H, --OH),
8.46(s, 1H, 2-H), 7.84(s, 1H, 5-H), 7.84-6.50(m, 4H,
2',4',5',6'-H), 7.20(s, 1H, 8-H), 3.96(s, 3H, --OCH.sup.3), 3.93(s,
3H --OCH.sub.3). GC/MS m/z: (C.sub.16H.sub.15N.sub.3O.sub.3.HCl) C,
H, N.
4-(4'-Hydroxyl-3'-Carboxyphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P182) Yield 79.25%; m.p. > 300.0.degree. C. .sup.-H
NMR(DMSO-d.sub.6)I: .delta. 10.53(s, 1H, --NH), 8.53(s, 1H, 2-H),
8.10-78.2(m, o3H, 2', 5', 6', --H), 7.26(s, 1H, 5-H), 6.9(s, 1H,
80H), 4.01(s, 3H, --OCH.sub.3), 3.99(s, 3H, --OCH.sub.3). UV(MeOH):
210.0, 239.0, 335.0 nm. IR(KBr).nu..sub.max: 3421, 2839, 1686,
1631, 1508, 1491, 1280 cm.sup.-1. GC/MS m/z 341(M.sup.+, 7.91),
323(M.sup.+ --OH, 12.19), 297(M.sup.+ --COOH, 12.35), 296(M.sup.+
--COOH - 1.1760), 295(M.sup.+ --COOH - 2, 28.65), 206(11.28).
4-(4'-Hydroxyl-3'-5'-dicholophenyl-6,7-dimethoxyquinazoline
(HI-P189) Yield 79.45%; m.p. 293.0-295.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): 11.32(s, 1H, --NH), 10.34(a, 1H, --OH), 8.87(s,
1H, 2-H), 8.29(s, 1H, 5-H), 7.90(s, 2H, 2',6'-H), 7.32(s, 1H, 8-H),
4.01(s, 3H, --OCH.sub.3), 3.99(s, 3H, --OCH.sub.3). UV(MeOH):
213.0, 232.0, 250.0, 335.0 nm. IR(KBr).nu..sub.max: 3479, 2564,
1641, 1579, 1429, 1282, 1147 cm.sup.-1. GC/MS m/z 367(M.sup.+ + 2;
66.57), 366(M.sup.+ + 1, 75.91), 365(M.sup.+, 100.00), 364(M.sup.+
- 1, 94.08), 349(M.sup.- OH, 11.16. Anal.
(C.sub.16H.sub.13Cl.sub.2N.sub.3O.sub.3) C, H, N. Found: C, 48.93;
H, 4.51; N, 10.00. --H.sub.1--Cl.sub.2N.sub.3O.sub.3.HCl requires:
C, 48.80; H, 4.31; N, 10.04%.
4-(4'-Bromo-6'-Hydroxymethylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P190) Yield 7o3.08%; m.p. 222.0-223.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.30(s, 1H, --OH), 8.22(s, 1H, --NH)O,
7.77.7.23(m, 5H, 5,8,2',3',5'-H), 4.49(s, 2H, PhCH.sub.2--H),
4.01(s, 3H, --OCH.sub.3), 3.90(s, 3H, --OCH.sub.3). UV(MeOH):
207.0, 250.0, 332.0 nm. IR(KBr).nu..sub.max: 3446, 2829, 2752,
1652, 1560, 1471, 1365, 1280 cm.sup.-1. GC/MS m/z 391(M.sup.- = 1,
29.33), 389(M.sup.-, 29.82), 360(M.sup.- CH.sup.2OH, 50.76),
358(52.39), 311(18.33), 280(43.20), 206(62.80), 191(100.00). Anal.
(C.sup.17H.sup.16BrN.sub.3O.sub.3.HCl) C, H, N.
4-(6'-Hydroxymethylphenyl)-amino-6,7-dimethosyquinazoline (HI-P191)
Yield 78.45%; m.p. 215.0-218.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.54(s, 1H, --NH)O, 8.70(s, 1H, 2-H),
8.34(s, 1H, 5-H), 7.62-7.33(m, 4H, 3',4',5',6'-H), 7.39(s, 1H,
8-H), 4.49(s, 2H, PhCH.sub.2OH), 3.99(s, 3H, --OCH.sub.3), 3.98(s,
3H, --OCH.sub.3). UV9MeOH): 209.0, 224.0, 246.0, 335.0 nm.
IR(KBr).nu..sub.max: 3421, 2941, 1675, 2606, '128, 1508, 1437o,
1244 cm.sup.-1. GC/MS m/z 311(M.sup.-, 38.07), 310(M.sup.- - 1,
27.04), 28o0(M.sup.- CH.sub.2OH, 100.00), 206(17.24), 191(51.34).
4-(2',4'-Dihydroxyphenyl)-amino-6,7-dimethoxyquinazoline (HI-P192)
Yield 86.25%; m.p. 240.0.degree. C.(dec). .sup.1H
NMR(DMSO-d.sub.6): 10.92(s, 1H, --NH), 976(s, 1H, --OH), 9.59(s,
1H, --OH), 8.67(s, 1H, 20H), 81.9(s, 1H, 8-H), 7.36(s, 1H, 50H),
705(d, 1H, J-8.7 Hz, 1'-H), 6.51(s, 1H, 5'-H), 6.31(d, 1H, J-8.7
Hz, 3'-H), 3.98(s, 6H, --OCH.sub.3). UV(MeOH): 206.0, 209.0, 223.0,
250.0, 342.0, 486 nm. IR(KBR).nu..sub.max: 3391, 3139, 2938, 2850,
1633, 1607, 1567, 1509, 1447, 1359, 1220, 1189, 1055 cm.sup.-1.
GC/MS m/z: 314(M.sup.- = 1, 13.00), 313(m.sup.-, 72.80),
312(m.sup.+ - 1, 10.20), 296(524), 206(17.50).
4-(2',3'-Dihydroxyphenyl)-amino-6,7-dimethoxyquinazoli- ne
(HI-P192) Yield 86.25%; m.p 240.0.degree. C.(dec). .sup.1H
NMR(DMSO-d.sub.6): 10.00(s, 1H, --NH), 9.37(s, 1H, --OH), 8.41(s,
1H, 2-H), 7.78(s, 1H, 5-H), 7.49(d, 1H, J.sub.2',3'=2.7 Hz, 2'-H),
7.55(dd, 1H, J.sub.5',6'=9.0 Hz, J.sub.2',6'=2.7 Hz, 6'-H), 7.16(s,
1H, 8-H), 6.97(d, 1H, J.sub.5',6'=9.0 Hz, 5'-h), 3.93(s, 3H,
--OCH.sub.3), 3.91(s, 3H, --OCH.sub.3). UV9MeOH): 209.0, 224.0,
249.0, 330.0 nm. IR(KBr).nu..sub.max: 3448, 2842, 1623, 1506, 1423,
1241 cm.sup.-1. GC/MS m/z: 341(M.sup.+, 100.00), 326(M.sup.-
CH.sub.3, 98.50), 310(M.sup.+ --OCH.sub.3, 12.5), 295(9.0),
189(13.5), 155(13.8). Found: C, 52.35; H, 4.16; Cl, 19.15; N,
11.39. C.sub.16H.sub.14ClN.sub.3O.sub.3HCl requires: C, 52.32; H,
4.09; Cl, 19.07; N, 11.44%.
4-(2',4'-Dihydroxyl-1',3'-diazine-5')-amino--
6,7-dime-thoxyquinazoline (HI-P215) (Yield 89.23%, m.p. >
300.0.degree. C.) .sup.1H NMR(DMSO-d.sub.6): .delta. 8.59(s, 1H,
2-H), 7.89(s, 1H, 5-H), 7.60(d, 1H, 6'-H), 7.09(s, 1H, 8-H),
3.78(s, 3H, --OCH.sub.3), 3.76(s, 3H, --OCH.sub.3). UV(MeOH):
222.0, 246.0, 331.0 nm. IR(KBr).nu..sub.max: 3446, 3212, 3057,
1750, 1682, 1620, 1590, 1511, 1420, 1265 cm.sup.-1. GC/MS m/z:
315(M.sup.- .57.52), 206(46.50), 191(18.21), 127(100.00).
4-(3'-Hydroxymethylphenyl)-am- ino-6,7-dimethoxyquina-zoline
(HI-P259) Yield 74.28%; m.p. 230.0-232.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.29(s, 1H, --NH), 8.83(s, 1H, 2-H)I,
8.28(s, 1H, 5-H), 7.61-7.25(m, 4H, 2',4',5',6'-H), 7.36(s, 1H,
8H)O, 4.57(s, 2H, --CH2OH), 4.02(s, 3H, --OCH.sub.3), 4.00(s, 3H,
--OC.sub.3). UV(MeOH): 207.0, 224.0, 251.0, 334.0 nm.
IR(KBr).nu..sub.max: 3500, 3029, 1639, 1589, 1514, 1456, 1284
cm.sup.-1. GC/MS m/z: 281(M.sup.- + 1 --CH.sub.2OH, 54.00),
280(M.sup.- --CH2OH, 100.00). Found: C, 58.68; H;, 5.30; N, 12.02.
C.sub.16H.sub.15N.sub.3O.sub.2.HCl requires: C, 58.79; H, 5.19; N,
12.10%. 4-[4'-(2"-Hydroxylethylphenyl)]-amino-6,7-dimethoxyqui-naz-
oline (HI-P265) Yield 92.30%; m.p. 235.0-240.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.44(s, 1H, --NH), 8.79(s, 1H, 2-H),
8.34(s, 1H, 5-h)I, 7.56(d, 2H, J=8.1 Hz, 2',6'-H), 7.34(d, 2H,
J-8.1 Hz, 3',5'-H), 7.31(s, 1H, 8H), 4.00(s, 3H, --OCH.sub.3),
3.99(s, 3H, --OCH.sub.3), 3.64(t, 2H, j=6.9 Hz, 1"-H)I, 2.77(t, 2H,
J=6.9 Hz, 2"-H). UV(MeOH): 204.0, 226.0, 251.0, 335.0 m.
IR(KBr).nu..sub.max: 3361, 3015, 27o6o7, 1628, 1581, 1514, 1432,
1282 cm.sup.-1. GC/MS m/z: 281(17.00), 253(10.00), 207(100.00).
4-[2'-(2"-Hydroxylethylphenyl)]-amino-6,7-dimethoxyqui-nazoline (HI
P266) Yield 87.69%; m.p/ 228.0-230.0.degree. C. .sup.1H
NMR-DMSO-d.sub.6): .delta. 11.32(s, 1H, --NH), 8.74(s, 1H, 2'-H),
8.13(s, 1H, 5-H), 7.46-7.34(m, 4H, 3',4',5,6'-H), 7.32(s, 1H, 8H),
4.00(s, 3H, --OCH.sub.3), 3.99(s, eH, --OCH.sub.3), 3.58(t, 2H,
J-7.2 Hz, 1"-H), 2.75(t, 2H, J=7.2 Hz, 2"-H). UV(MeOH): 210.0,
226.0, 249.0, 332.0 nm. IR(KBr).nu..sub.max: 3366, 3226, 3056,
2917o, 2685, 21638, 1571, 1514, 1467, 1277 cm.sup.-1. GC/MS m/z:
281(20.00), 253(9.00), 207(100.00).
4-(1'-Naphthol-4')-amino-6,7-edimethoxyqui- nazoline (HI-P274)
Yield 79.26; m.p. 205.0-208.0.degree. C. .sup.1H NMR-DMSO-d.sub.6):
.delta. 11.64(s, 1H, --NH), 10.61(s, 1H, --OH), 8.59(s, 1H, 2-h),
8.41(s, 1H, 5-H), 8.22-6.98(m, 5H, 3',5',6',7',8'-H), 7.40(s, 1H,
8H), 4.00(s, 3H, --OCH.sub.3), 3.99(s, 3H, --OCH.sub.3). UV9MeOH):
208.0, 215.0, 225.0, 240.0, 330.0 nm. IR(KBr).nu..sub.max: 3438,
3211, 3061, 2932, 2834, 1633, 1576, 1509, 1437, 1380, 1276, 1215
cm.sup.-1. GC/MS m/z: 281(51.00), 253(22.00), 207(88.00). Found: C,
62.26; H, 4.87; N, 10.77. C.sub.20H.sub.17N.sub.3O.sub.3.HCl
requires: C, 62.66; H, 4.70; N, 10.96%.
4-(2'-Naphthol-1')-amino-6,7-dimethoxyquinazoline (HI-P275) Yield
83.17%; m.p. 218.0-220.0.degree. C. .sup.1H NMR(DMSO-d.sub.6):
.delta. 11.33(s, 1H, --NH), 10.22(s, 1H, --OH), 8.62(s, 1H, 2-H),
8.40(s, 1H, 5-H), 7.98-7.31(m, 6H, 3',4',5',6',7"8'-H), 7.41(s, 1H,
8H), 4.02(s, 3H, --OCH.sub.2), 4.00(s, 3H, --OCH.sub.3), .
UV(MeOH): 206.0, 210, 219.0, 225.0, 230.0, 340.0 nm.
IR(KBr).nu..sub.max: 3391, 3165, 3051, 2938, 2840, 1628, 1576,
1504, 1437, 1281, 1215 cm.sup.-1. GC/MS m/z: 348(M.sup.- +1, 7.00),
347(M.sup.-, 100.00), 346(M.sup.- 1.22.00), 331(15.00), 330(12.00),
281(23.00), 253(12.00), 207(49.00). Found: C, 62.91; H, 4.76; N,
10.75. C.sub.20H.sub.1N.sub.3O.sub.3.HCl requires: C, 62.66; H, 4,
70; N, 10.96%. 4-[3'-(1"-Hydroxyethyl)]-amino-6,7-dim-
ethoxyquinazoline (HI-P276) Yield 79.21%; m.p. 215.0-218.0.degree.
C. .sup.1H NMR(DMSO-d.sub.6): .delta. 11.40(s, 1H, --NH), 8.81(s,
1H, 20H), 8.31(s, 1H, 5-H)O, 7.60-7.26(m, 4H, 2',4',5',6'-H),
7.41(s, 1H, 8H), 4.65(q, 1H, J=6.6 Hz, --CH(OH)CH.sub.3), 4.00(s,
3H, --OCH.sub.3), 3.98(s, 3H, --OCH.sub.3), 1.350(d, 3H, J=6.6 Hz,
--CH.sub.3). UV9MeOH): 204.0, 216.0, 220.0, 224.0, 250.00, 348.0
nm. IR(KBr).nu..sub.max: 3407, 3030, 2977, 2840, 1643, 1591 1514,
1463, 1370, 1282, 1230 cm.sup.-1. GC/MS m/z: 325(M.sup.- + 1,
67.00), 324(M.sup.-, 100.00), 323(M.sup.- 1.22.00), 308(17.00),
307(56.00), 306(21.00), 281(2.00), 280(8.00), 264(6.00).
4-(4'-Hydroxy-3',5'-diphenylphenyl)-amino-6,7-dime-hoxyquinazoline
(HI-P277) Yield 76.11%; m.p. 255.0-257.0.degree. C. .sup.1H
NMR_DMSO-d.sub.6): .delta. 11.50(s, 1H, --NH), 8.80(d, d, 2H,
2',6'-H), 8.58(s, 1H, 5-H), 7.60-7.30(m, 10H, 3', 5', Ph--H),
7.39(s, 1H, 8H), 4.00(s, 3H, --OCH.sub.3), 3.97(s, 3H,
--OCH.sub.3), 1.350(d, eH, J=6.6 Hz, --CH.sub.3). UV(MeOH): 210.0,
214.0, 229.0, 239.0, 345.0, 248.0, 352.0 nm. IR(KBr).nu..sub.max:
3520, 3218, 3023, 2935, 1630, 1562, 1518, 1457, 1281, 1234
cm.sup.-1. GC/MS m/z: 281(35.00), 267(6.00), 253(10.00),
207(100.00).
4-(4'-Hydroxyl-2'-chlorophenyl)-amino-6,7-dimethoxy-quinazoline
(HI-P2878) Yield 81.44%; m.p. 245.0-247.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.39(s, 1H, --NH)O, 10.30(s, 1H, --OH),
8.75(s, 1H, 2-H), 8.24(s, 1H, 5-H), 7.38-6.85(m, 3H, 3',5',6'-H),
7.37(s, 1H, 8H), 3.98(s, 3H, --OCH.sub.3), 3.96(s, H.sub.3,
--OCH.sub.3). UV(MeOH): 222.0, 234.0, 239.0, 245.0, 254.0, 348.0
nm. (R(KBr).nu..sub.max: 3448, 3242, 3144, 3025, 2917, 2834,
1638,
1591, 1514, 1437, 1365, 1277, 1209 cm.sup.-1. GC/MS c/z:
332(M.sup.- + 1, 5.00), 331(M.sup.-, 17.00), 330(M.sup.- - 1,
5.00), 297(17.00), 296(100.00), 281(18.00), 280o(29.00), 253(9.00).
4-(2'-Hydroxy-naphthyl-3')-amino-6,7-dimethoxyquinazolin (HI-P292)
Yield 87.41%; m.p. 277.0-279.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.38(s, 1H, --NH)O, 10.35(s, 1H, --OH),
8.73(s, 1H, 2-H), 8.25(s, 1H, 5-H), 7.93-7.30(m, 6H,
1',4',5',6',7',8'-H), 7.37(s, 1H, 8H)O, 4.00(s, 6H, --OCH.sub.3).
UV(MeOH): 204.0, 221.0, 224.0, 230.0, 256.0, 344.0 nm.
IR(KBr).nu..sub.max: 3479, 3386, 3036, 2901, 1632, 1581, 1504,
1437, 1281 cm.sup.-1. GC/MS m/z: 281(41.00), 253(11.00),
207(100.00). Found: C, 62.87; H;, 4.83; N, 1o0.78.
C.sub.20H.sub.1N.sub.3O.sub.3.HCl requires: C, 62.66; H, 4.70, N,
10.96%. 4-(1'-Hydroxy-naphthyl-5')-amino-6,70-dimethoxyquina-zoline
(HI-P293) Yield 87.21%; m.p. 204.0-205.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.73(s, 1H, --NH), 10.43(s, 1H, --OH),
8.65(s, 1H, 2-H, 8.38(s, 1H, 5-H), 8.21-6.95(m, 6H,
2',3',4',6',7',8'-H), 7.33(s, 1H, 8H)O, 4.00(s, 6H, --OCH.sub.3).
UV9MeOH): 204.0, 214.0, 224.0, 229.0, 235.0 348 nm.
IR(KBr.nu..sub.max: 3449, 3335, 3102, 2927o, 1633, 1571, 1509,
1437, 1287 cm.sup.-1. Found: C, 62.23; H, 4.96; N, 10.89.
C.sub.20H.sub.17N.sub.3O.sub.3.HCl requires. C, 62.66; H, 4.70; N,
10.96%. 4-(4'-Hydroxy-3,5-diiodophenyl)-amino-6-
,7-dimethoxy-quinazoline (HI-P294) Yield 77.47& m.p.
259.0-260.0.degree. C. .sup.1H NMR(DMSO-d.sub.6): .delta. 11.13(s,
1H, NH), 9.73(s, 1H, --OH), 8.87(s, 1H, 2-H), 8.16(s, 1H, 5-H),
8.09(s, 2H, 1',6'-H), 7.28(s, 1H, 8H), 3.98(s, 6H, --OCH.sub.3),.
UV(MeOH).lambda..sub.max: 217.0, 227.0, 252.00 nm.
IR(KBr.nu..sub.max: 3457, 3201, 2934, 2832, 2566, 1629, 1562, 1521,
1439, 1275, 1075 cm.sup.-1. GC/MS m/z: GC/MS m/z 422(M.sup.-
1.33.53), 405(7.50), 281(86.67), 221(51.80), 207(91.30). Found: C,
32.60; H, 2.50; N, 6.92. C.sub.16H.sub.13I.sub.2N.sub.3O.sub.3.HCl
requires: C. 32/82.' J. 2.39; N, 7.18%.
4-(4'-Hydroxy-3'-iodophenyl)-amino-6,7-- dimethoxyquinazoline
(HI-P299) Yield 71.59%; m.p. 248.0-250.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.32(d, 1H, NHO), 10.62(s, 1H, --OH,
8.79(s, 1H, 2-H), 8.26(s, 1H, 5-H), 7.98-6.98(m, 3H, 2',3',6'-H),
7.32(s, 1H, 8H), 3.98(s, 3H, --OCH.sub.3), 3.97(s, 3H,
--OCH.sub.3). UV(MeOH).lambda..sub.max(.- epsilon.): 217.0, 227.0,
252.0 nm. IR(KBr).nu..sub.max: 3411, 2975, 2730, 2366, 1634, 1573,
1501, 1429, 1229, 1075 cm.sup.-1. GC/MS m/z: 406(M.sup.- 1.3.33),
405(M.sup.- 2, 7.50), 281(M.sup.+ - 1 --I, 26.67), 253(11.80),
207(100.00). Found: C, 41.96; H, 3.40; N, 8.98.
C.sub.16H.sub.14IN.sub.3O.sub.3.HCl requires: C, 41.83; H, 3.26; N,
9.15%.
[0104]
5TABLE 5 Fluoroquinazoline Derivatives 72 (HI-P352) (HI-P353) No R
Formular MW HI-P144 2-F, 3-F, 5-F, 6-F, 4-Br
C.sub.16H.sub.10BrF.sub.4N.sub.3O.sub.2 432 HI-P214 2-F, 4-Br
C.sub.16H.sub.13BrFN.sub.3O.sub.2 378 HI-P218 3-CF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.2 349 HI-P219 4-OCF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 HI-P221 4-F
C.sub.16H.sub.14FN.sub.3O.sub.2 299 HI-P223 4-CF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.2 349 HI-P224 3-F
C.sub.16H.sub.14FN.sub.3O.sub.2 299 HI-P228 2-CF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.2 349 HI-P232 4-SO.sub.2F
C.sub.16H.sub.14FN.sub.3O.sub.4S 363 HI-P264 2-F
C.sub.16H.sub.14FN.sub.3O.sub.2 299 HI-P352 *
C.sub.25H.sub.20F.sub.6N.sub.4O.sub.2 522 HI-P353 *
C.sub.25H.sub.20F.sub.6N.sub.4O.sub.2 522 HI-P364 3-OCF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 HI-P365 2-OCF.sub.3
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 HI-P366 3-CF.sub.3,
5-CF.sub.3, C.sub.18H.sub.13F.sub.6N.sub.3O.sub.2 417 HI-P367
2-CF.sub.3, 5-CF.sub.3, C.sub.18H.sub.13F.sub.6N.sub.3O.sub.2 417
HI-P369 3-F, 4-OH C.sub.16H.sub.14FN.sub.3O.sub.3 315 HI-P408 3-F,
5-F, 4-OH C.sub.16H.sub.13F.sub.2N.sub.3O.sub.3 333 73 HI-P352 74
HI-P353
Example 6
[0105]
6 Fluorine Substituted Quinazoline Compounds Fluorine substituted
quinazoline derivatives were synthesized and characterized as
discussed above for Example 1. The structures and physical data are
shown below: No Name Structure Formula MW 1 P-144 75
C.sub.16H.sub.10BrF.sub.4N.sub.3O.sub.2 432 2 P-214 76
C.sub.16H.sub.13BrFN.sub.3O.sub.2 378 3 P-218 77
C.sub.17H.sub.13F.sub.4N.sub.3O.sub.2 367 4 P-219 78
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 5 P-221 79
C.sub.16H.sub.14FN.sub.3O.sub.2 299 6 P-223 80
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.2 349 7 P-224 81
C.sub.16H.sub.14FN.sub.3O.sub.2 299 8 P-228 82
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.2 349 9 P-232 83
C.sub.16H.sub.14F.sub.2SN.sub.3O.sub.4 363 10 P-264 84
C.sub.16H.sub.14FN.sub.3O.sub.2 299 11 P-352 85
C.sub.25H.sub.20F.sub.6N.sub.4O.sub.2 522 12 P-353 86
C.sub.25H.sub.20F.sub.6N.sub.4O.sub.2 522 13 P-364 87
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 14 P-365 88
C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3 365 15 P-366 89
C.sub.18H.sub.13F.sub.6N.sub.3O.sub.2 417 16 P-367 90
C.sub.18H.sub.13F.sub.6N.sub.3O.sub.2 417 17 P-369 91
C.sub.16H.sub.14FN.sub.3O.sub.3 315 18 P-408 92
C.sub.16H.sub.13F.sub.2N.sub.3O.sub.3 333
4-(2',3',5',6'-Terrafluoro-4'-bromophenyl)-amino-6,7-dime-thoxyquinazolin-
e (HI-P144) The yield 78.24%: m.p. 180.0-182.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 7.78(s, 1H, 2-H), 7.53(s, 1H, 5-H),
6.79(s, 1H, 8-H), 3.81(s, 3H, --OCH.sub.3), 3.3.79(s, 3H,
--OCH.sub.3). Found: C, 41.12; H, 2.41; N, 9.89,
C.sub.10H.sub.10BrF_N.sub.3O.sub.- 2.HCl. requires: C, 41.11; H,
2.36; N, 9.93%.
4-(2'-Fluoro-4'-bromophenyl)-amino-6,7-dimethoxyquina-zoline
(HI-P214) The yield 77.21%; m.p. 247.0-252.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 8.57(s, 1H, 2-H), 7.91(s, 1H, 5-H),
7.57(d, 1H, 3'-H), 7.34(m, 2H, 5',6'-H), 7.07(s, 1H, 8-H), 3.78(s,
3H, --OCH.sub.3), 3.77(s, 3H, --OCH.sub.3). UV(MeOH): 204.0, 215.0,
250.0, 330.0 nm. IR(KBr).nu..sub.max: 3431, 2629, 1633, 1580, 1511,
1420, 1278 cm.sup.-1. GC/MS m/z 379(M.sup.+ + 1, 34.39),
378(M.sup.-, 31.33), 377(M.sup.- - 1, 39.08), 360(62.05),
359(31.58), 358(62.57), 357(19.81), 299(19.31), 298(100.00),
282(17.88), 240(28.76).
4-(3'-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinaz- o-line
(HI-P218) The yield 85.61%: m.p. 242.0-245.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.09(s, 1H, --NH), 8.67(s, 1H, 2-H),
8.03(s, 1H, 5-H), 7.92-7.43(m, 4H, 2'4'5',6'-H), 7.10(s, 1H, 8-H),
3.81(s, 3H, --OCH.sub.3), 3.79(s, 3H, --OCH.sub.3). UV(MeOH):
206.0, 276.0, 349.0 nm. IR.nu..sub.max(KBr): 3372, 3257, 2935,
1626, 1512, 1380, 1225 cm.sup.-1. GC/MS m/z 350(M.sup.+ + 1, 10.5),
249(M.sup.-, 85.5), 173(M.sup.- - 1, 100.0), 332(10.5), 290(8.8).
4-(4'-Trifluoromethoxylphenyl)-amino-6,7-dimethoxyqui-nazoline
(HI-P219) The yield 83.14%; m.p. 228.0-230.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.39(s, 1H, --HN), 8.63(s, 1H, 2-H),
8.18(s, 1H, 5-H), 7.63(t, 2H, 3',5'-H), 7.27(t, 2H, 2',6'-H),
7.15(s, 1H, 8-H), 3.81(s, 3H, --OCH.sub.3), 3.78(s, 3H,
--OCH.sub.3). UV(MeOH): 209.0, 216.0, 251.0, 332.0 nm.
IR(KBr).nu..sub.max: 3207, 2839, 2762, 1633, 1508, 1480, 1276
cm.sup.-1. GC/MS m/z 366(M.sup.+ + 1, 12.50), 365(M.sup.-, 75.00),
364(M.sup.- - 1, 100.00), 348(17.50), 319(19), 306(8.00),
207(15.00). 4-(4'-Fluorophenyl)-amino-6,7-dimethoxyquinazoline
(HI-P221) The yield 84.25%: .sup.1H NMR(DMSO-d.sub.6): .delta.
11.19(s, 1H, --HN), 8.60(s, 1H, 2-H), 8.08(s, 1H, 5-H), 7.50(t, 2H,
3'-H), 7.13(s, 1H, 8-H), 7.12(t, 2H, 2',6'-H), 3.79(s, 3H,
--OCH.sub.3), 3.78(s, 3H, --OCH.sub.3). UV(MeOH): 225.0, 251.0,
333.0 nm. IR(KBr).nu..sub.max: 3205, 3007, 2837, 1633, 1580, 1508,
1470, 1220 cm.sup.-1. GC/MS m/z 300(M.sup.+ + 1, 10.76),
299(m.sup.-, 76.92), 39.8(M.sup.- - 1, 100.00), 282(20.00),
253(13.08), 207(3.80). Found: C, 57.17; H, 4.37; N, 12.47,
C.sub.16H.sub.14FN.sub.3O.sub.2.HCl requires C, 57, 31; H, 4.48; N,
12.54%. 4-(4'-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P223) The yield 91.70%: m.p. 243.0-245.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.47(s, 1H, --NH), 8.67(s, 1H, 2-H),
8.23(s, 1H, 5-H), 7.79(d, 2H, J=8.4 Hz, 3'5'-H), 7.61(d, 2H, J=8.4
Hz, 2'6'-H), 7.17(s, 1H, 8-H), 3.82(s, 3H, --OCH.sub.3), 3.78(s,
3H, --OCH.sub.3). GC/MS m/z 350(M.sup.- + 1, 11.00), 349(M.sup.-,
65.00), 348(M.sup.- - 1, 100.00), 332(18.50), 303(10.00),
207(18.50). Found: C, 53.01; H, 3.94; N, 10.88.
C.sub.1--H.sub.14F.sub.3N.sub.- 3O.sub.2HCl requires C, 52.98; H,
3.90; N, 10.91%. 4-(4'-Flurophenyl)-amino-6,7-dimethoxyquinazoline
(HI-P224) The yield 88.69%; m.p. 254.0-255.0 0.degree. C. .sup.1H
NMr(DMSO-d.sub.6): .delta. 11.16(s, 1H, --HN), 8.67(s, 1H, 2-H),
8.09(s, 1H, 5-H), 7.13(s, 1H, 8-H), 7.51-6/94(m, 4H,
2',3',5',6'-H)O, 3.80(s, 3H, --OCH.sub.3), 3.79(s, 3H,
--OCH.sub.3). UV(MeOH): 206.0, 226.0, 251.0, 333.0, 343 nm.
IR(KBr).nu..sub.max: 3437, 3211, 2619, 1637, 1580, 1500, 1448, 1281
cm.sup.-1. GC/MS m/z (300(M.sup.+ + 1, 8.00), 299(M.sup.-, 68.00),
398(M.sup.- 1, 100.00), 282(21.60), 253(25.00), 207(80.00),. Found:
C, 57.25; H, 4.58; N, 12.42. C.sub.16H.sub.14FN.sub.3O.sub.2.Hcl
requires C, 57.31; H, 4.48; N, 12.54%.
4-(2'-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinazolin- e
(HI-P228). The yield 83.57%; m.p. 242.0-245.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.58(s, 1H, --HN), o8.76(s, 1H, 2-H),
8.25(s, 1H, 5-H), 7.95-7.62(m, 4H, 3',4',5',6'-H), 7.38(s, 1H, 8H),
4.01(S, 3H, --OCH.sub.3), 3.00(s, 3H, --OCH.sub.3). GC/MS m/z
350(M.sup.- + 1, 8.50), 349(M.sup.-, 32.00), 348(M.sup.+ -
1.31.50), 328(18.50), 207(5.0)I, 280(M.sup.+ --CF.sub.3, 100.00),
264(18.50), 207(32.50). Found: C, 52.71; H, 4.26; N, 10.91%.
4-[4'-benzenesulfanilyl fluoride]-amino-6,7-dimethoxyquinazoline
(HI-P232) Yield 84.02%; m.p. 228.0-230.0.degree. C. .sup.1H
NMR9DMSO-d.sub.6): .delta. 11.62(s, 1H, --HN), 8.78(s, 1H, 2-H),
8.29(s, 1H, 5-H), 8.12-8.02(m, 4H, 2",3",5",6"-H), 7.21(s, 1H,
8-H), 3.86(s, 3H, --OCH.sub.3), 3.81(s, 3H, --OCH.sub.3). UV(MeOH):
208.0, 215.0, 253.0, 278.0, 338.0 nm. IR(KBr).nu..sub.max: 3440,
3277, 2571, 1635, 1580, 1516, 1435, 1209 cm.sup.-1. GC/MS m/z:
281(43.00), 253(12.00), 207(100.00). Found: C, 48.13; H, 3.73; N,
10.53. C.sub.16H.sub.14FN.sub.3O.sub.4S.HCl requires: C, 48.12; H,
3.76; N, 10.53%. 4-(2'-Fluorophenyl)-amino-6,7-dimethoxyquinazoline
(HI-P264) Yield 73.58%; m.p. 233.0-235.0 0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.69(d, 1H, --NH), 8.82(s, 1H, 2-H),
8.37(s, 1H, k 50H), 7.59-7.32(m, 4H 3',4'5',6'-H), 7.41(s, 1H,
8H)O, 4.02(s, 3H, --OCH.sub.3), 4.01(s, 3H, --OCH.sub.3). UV(MeOH):
204.0, 226.0, 248.0, 330.0 nm. IR(KBr.nu..sub.max: 3454, 3032,
2638, 1630, 1589, 1514, 1430, 1291 cm.sup.-1. GC/MS m/z 300(M.sup.+
= 1, 7.00), 299(M.sup.-, 38.00), 298(M.sup.- 1.22.00), 280(M.sup.-
F, 100.00), 264(15.00), 207(35.00). Found: C, 57.12; H, 4.57; N,
12.45. C.sub.16H.sub.14FN.sub.3O.sub.2.HCl requires: C, 57.31; H,
4.48; N, 12.54%.
4-{4'-[2"-(4'''-Aminophenyl)-hexafluoropropyl]phenyl}-amin-
o-6,7-dimethoxyquinazoline (HI-P352) Yield, 80.41%, m.p.
280.0-282.0.degree. C. .sup.1H NMR(DMSO-d.sub.6): .delta. 11.87(s,
1H, --NH), 8.91(s, 1H, 2-H)I, 8.55-7.18(m, 10H,
5,8,2',3',5',6',2''',3''',5''',6'''-H), 4.05(s, 3H, --OCH.sub.3),
4.00(s, 3H, --OCH.sub.3). .sup.19F NMR(DMSO-d.sub.6): 128.76.
Found: C, 50.33; H, 3.87; N, 9.57.
C.sub.25H.sub.20F.sub.6N.sub.4O.sub.2.- 2HCl requires: C, 50.50; H,
3.70; N, 9.42% 4-{3'-[2"-(3'''-Aminophe-
nyl)-hexafluoropropyl]phenyl}-amino-6,7- dimethoxyquinazoline
(HI-P353) Yield, 83.11%, m.p. 292.0-284.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.68(s, 1H, --NH), 8.81(s, 1H, 2-H),
8.44-7.09(m, 10H, 5,8,2',4',5',6',2''',4''',5'''6'''-H). 4.00(s,
3H, --OCH.sub.3), 3.97(s, 3H, --OCH.sub.3). .sup.19F
NMR(DMSO-d.sub.6): 129.21. Found: C, 53.96; H, 3.93; N, 9.77.
C.sub.25H.sub.20F.sub.6- N.sub.4O.sub.2.HCl requires: C, 53.76; H,
3.76; N, 10.03%
4-(3'-Trifluoromethoxylphenyl)-amino-6,7-dimethoxyquinazoline
(HI-P364) Yield, 83.25%. m.p. 233.0-235.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.65(s, 1H, --NH), 8.88(s, 1H, 2-H),
8.41(s, 1H, 5-H), 7.86-7.29(m, 4H, 2',4',5',6'-H), 7.36(s, 1H,
8-H), 4.02(s, 3H, --OCH.sub.3). 3.98(s, 3H, --OCH.sub.3). .sup.19F
NMR(DMSO-d.sub.6): 135.37. GC/MS m/z: 366(M.sup.+ + 1, 11.0),
365(M.sup.+, 67.0), 364(M.sup.+ - 1, 100.0). Found: C, 50.93; H,
3.75; N, 10.61. C.sub.17H.sub.14F.sub.3N.sub.3O.sub.3.HCl requires:
C, 50.97; H, 3.74; N, 10.47%.
4-(2'-Trifluoromethoxylphenyl)-amino-6,- 7-dimethoxyquinazoline
(HI-P365) Yield, 77.85%. m.p. 235.0-237.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.68(s, 1H, --NH), 8.80(s, 1H, 2-H),
8.32(s, 1H, 5-H), 7.64-7.53(m, 4H, 3',4',5',6'-H), 7.40(s, 1H,
8-H), 3.99(s, 6H, --OCH.sub.3). .sup.19F NMR(DMSO-d.sub.6): 135.71.
GC/MS m/z: 366(M.sup.- + 1, 2.0), 365(M.sup.+, 15.0), 364(M.sup.+ -
1, 4.0), 281(21.0), 280(M.sup.- --OCF.sub.3 100). Found: C, 50.83;
H, 3.79; N, 10.52. C.sub.1--H.sub.14F.sub.3N.sub.3O.sub.3.HCl
requires: C, 50.87; H, 3.74; N, 10.47%.
4-(3',5'-Ditrifluoromethylphenyl)-amno-6,7-dimethoxyqui- nazoline
(HI-P366) Yield, 82.88% m.p. 270.0-272.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.87(s, 1H, --NH), 8.97(s, 1H, 2-H),
8.60)s, 2H, 2',6'-H), 8.43(s, 1H, 5-H), 7.98(s, 1H, 4'-H), 7.35(s,
1H, 8-H), 4.03(s, 3H, --OCH.sub.3). 3.99(s, 3H, --OCH.sub.3).
.sup.19F NMR(DMSO-d.sub.6): XX GC/MS m/z: 418(M.sup.- + 1, 19.0),
417(M.sup.-, 100.0), 416(M.sup.- - 1, 73.0), 398(M.sup.- --F,
16.0), 398(M.sup.- --F, 16.0), 348(M.sup.- --CF.sub.3, 16.0).
Found: C, 47.78; H, 3.20; N, 9.26.
C.sub.18H.sub.13F.sub.6N.sub.3O.sub.2.HCl requires: C, 47.68; H,
3.09; N, 9.27%. 4-(4'-Hydroxyl-3'-fluorophe-
nyl)-amino-6,7-dimethoxyquinazoline (HI-P369) Yield, 84.28%. m.p.
268.0-270.0.degree. C. .sup.1H NMR(DMSO-d.sub.6: .delta. 11.36(s,
1H, --NH), 10.13(s, 1H, --OH), 8.80(s, 1H, 2-H), 8.30(s, 1H, 5-H),
7.60-7.02(m, 3H, 2',5',6'-H), 7.31(s, 1H, 8-H), 3.99(s, 3H,
--OCH.sub.3), 3.97(s, 3H, --OCH.sub.3). .sup.19F NMR(DMSO-d.sub.6):
.delta. 57.38. Found: C, 54.90; H, 4.28; N, 11.91.
C.sub.16H.sub.14FN.sub.- 3O.sub.3.HCl requires C, 54.70; H, 4.27;
N, 11.97%.
4-(4'-Hydroxyl-3',5'-difluorophenyl)-amino-6,7-dimethoxy-quinazoline
(HI-P408) Yield, 83.15%, m.p. 228.0-230.0.degree. C. .sup.1H
NMR(DMSO-d.sub.6): .delta. 11.46(s, 1H, --NH), 10.39(s, 1H, 2-H),
8.36(s, 1H, 5-H), 7.56, 7.54 (s, s, 2H, 2',6'-H), 7.33(s, 1H, 8-H),
4.00)s, 3H, --OCH.sub.3), 3.98(s, 3H, --OCH.sub.3). .sup.19F
NMR(DMSO-d.sub.6: .delta. 60.25, 60.22. Found: C, 52.04; H, 4.17;
N, 11.10. C.sub.16H.sub.13F.sub.2N.sub.3O.sub.3.HCl. requires C,
52.03; H, 3.79; N, 11.38%.
Example 7
Anti-Tumor Activities of Specific Quinazoline Compounds
[0106] The cytotoxicity of the substituted quinazoline derivative
compounds against a variety of human tumor cells was evaluated. The
relative importance of particular substituent groups on the
compounds was also studied. The substituted quinazoline derivative
compounds, prepared as described above, were tested, along with
DMSO as a control.
[0107] Cytotoxicity Assay
[0108] The cytotoxicity assay of various compounds against human
tumor cell lines was performed using the MTT
(3-[4,5-dimethylthiazol-2-yl]-2,5-- diphenyl tetrazolium bromide)
assay (Boehringer Mannheim Corp., Indianapolis, Ind.). Briefly,
exponentially growing tumor cells were seeded into a 96-well plate
at a density of 2.5.times.10.sup.4 cells/well and incubated for 36
hours at 37.degree. C. prior to drug exposure. On the day of
treatment, culture medium was carefully aspirated from the wells
and replaced with fresh medium containing the quinazoline compounds
at concentrations ranging from 0.1 to 250 .mu.M. Triplicate wells
were used for each treatment.
[0109] Human cell lines were obtained from American Type Culture
Collection (Rockville, Md.) and maintained as a continuous cell
line in Dulbecco's modified Eagles' medium supplemented with 10%
fetal bovine serum and antibiotics. Cells used in this study
include human leukemia cells (NALM-6 and MOLT-3), human breast
cancer cells (BR20), human prostate cancer cells (PC3), and human
brain tumor cells (U373).
[0110] The cells were incubated with the various compounds for
24-36 hours at 37.degree. C. in a humidified 5% CO.sub.2
atmosphere. To each well, 10 .mu.l of MIT (0.5 mg/ml final
concentration) was added and the plates were incubated at
37.degree. C. for 4 hours to allow MTT to form formazan crystals by
reacting with metabolically active cells. The formazan crystals
were solubilized overnight at 37.degree. C. in a solution
containing 10% SDS in 0.01 M HCl. The absorbance of each well was
measured in a microplate reader (Labsystems) at 540 nm and a
reference wavelength of 690 nm. To translate the OD 540 values into
the number of live cells in each well, the OD 540 values were
compared to those on standard OD 540-versus-cell number curves
generated for each cell line. The percent survival was calculated
using the formula: 1 % Survival = live cell number [ test ] live
cell number [ control ] .times. 100
[0111] The IC.sub.50 values were calculated by non-linear
regression analysis.
[0112] Detection of Apoptosis
[0113] The demonstration of apoptosis was performed by the in situ
nick-end-labeling method using ApopTag in situ detection kit
(Oncor, Gaithersburg, Md.) according to the manufacturer's
recommendations. Exponentially growing cells were seeded in 6-well
tissue culture plates at a density of 50.times.10.sup.4 cells/well
and cultured for 36 hours at 37.degree. C. in a humidified 5%
CO.sub.2 atmosphere. The supernatant culture medium was carefully
aspirated and replaced with fresh medium containing unconjugated
EGF or EGF-P154 at a concentration of 10, 25, or 50 .mu.g/ml. After
a 36 hour incubation at 37.degree. C. in a humidified 5% CO.sub.2
incubator, the supernatants were carefully aspirated and the cells
were treated for 1-2 minutes with 0.1% trypsi The detached cells
were collected into a 15 mil centrifuge tube, washed with medium
and pelleted by centrifugation at 1000 rpm for 5 minutes. Cells
were resuspended in 50 .mu.l of PBS, transferred to poly-L-lysine
coated coverslips and allowed to attach for 15 minutes. The cells
were washed once with PBS and incubated with equilibration buffer
for 10 minutes at room temperature.
[0114] After removal of the equilibration buffer, cells were
incubated for 1 hour at 37.degree. C. with the reaction mixture
containing terminal deoxynucleotidyl transferase (TdT) and
digoxigenin-11-UTP for labeling of exposed 3'-hydroxyl ends of
fragmented nuclear DNA. The cells were washed with PBS and
incubated with anti-digoxigenin antibody conjugated to FITC for 1
hour at room temperature to detect the incorporated dUTP. After
washing the cells with PBS, the coverslips were mounted onto slides
with Vectashield containing propidium iodide (Vector Labs,
Burlingame, Calif.) and viewed with a confocal laser scanning
microscope. Non-apoptotic cells do not incorporate significant
amounts of dUITP due to lack of exposed 3-hydroxyl ends, and
consequently have much less fluorescence than apoptotic cells which
have an abundance of exposed 3'-hydroxyl ends. In control
reactions, the TdT enzyme was omitted from the reaction
mixture.
[0115] Results:
[0116] The cytotoxicity results for each tested group of compounds
is shown in Tables 1-5 below:
7TABLE 1 Cytotoxic Activity of Bromo Substituted Quinazoline
Compounds against Leukemic (NALM-6 & MOLT-3) and Breast Cancer
(BT-20) NALM-6 MOLT-3 BT20 IC50 IC50 IC50 Drug (.mu.M) (.mu.M)
(.mu.M) HI-P79 142.1 194.9 201.5 HI-P88 >250 >250 >250
HI-P97 >250 >250 26.1 HI-P111 200.6 >250 >250 HI-P154
12.5 9.1 >250 HI-P160 135.2 240.7 25.5 HI-P164 >250 >250
39.2 HI-P190 >250 >250 >250 HI-P210 >250 >250
>250 HI-P211 >250 >250 >250 HI-P212 52.7 54.5 >250
HI-P214 >250 >250 >250 Hl-P222 34.0 48.3 >250 HI-P234
>250 >250 >250 HI-P241 >250 >250 >250 HI-P258
>250 >250 >250 HI-P260 32.4 51.3 82.1 HI-P261 72.6 148.5
218.6 HI-P262 >250 >250 >250
[0117]
8TABLE 2 Cytotoxic Activity of Chloro Substituted Quinazoline
Compounds against Leukemic (NALM-6 & MOLT-3) and Breast Cancer
(BT-20) NALM-6 MOLT-3 BT20 IC50 IC50 IC50 Drug (.mu.M) (.mu.M)
(.mu.M) HI-P87 95.9 >104.6 >250 HI-P93 >250 >250
>250 HI-P189 >250 >250 >250 HI-P197 39.3 68.0 136.9
HI-P239 29.6 28.7 25.7 HI-P246 >250 >250 >250 HI-P268
215.2 227.4 121.5 HI-P269 >250 >250 >250 HI-P415 67.9
>250 38.1
[0118]
9TABLE 3 Cytotoxic Activity of Iodide Substituted Quinazoline
Compounds against Leukemic (NALM-6 & MOLT-3), Breast Cancer
(BT-20) and Brain Tumor (U373) cells NALM-6 MOLT-3 BT20 U373 IC50
IC50 IC50 IC50 Drug (.mu.M) (.mu.M) (.mu.M) (.mu.M) HI-P270 >250
78.9 >250 >250 HI-P271 6.1 9.6 >250 >250 HI-P294
>250 >250 >250 >250 HI-P299 15.4 60.1 >250 >250
HI-P300 58.0 59.1 72.6 116.2
[0119]
10TABLE 4 Cytotoxic Activity of OH Substituted Quinazokline
Compounds against Leukemic (NALM-6 & MOLT-3), Breast Cancer
(BT-20) and Brain Tumor (U373) cells NALM-6 MOLT-3 BT20 U373 IC50
IC50 IC50 IC50 Drug (.mu.M) (.mu.M) (.mu.M) (.mu.M) HI-P93 >250
>250 >250 >250 HI-P97 >250 >250 26.1 161.2 HI-P131
32.1 38.6 >250 >250 HI-P154 12.5 9.1 >250 167.4 HI-P189
>250 >250 >250 >250 HI-P190 >250 >250 >250
>250 HI-P192 >250 >250 >250 >250 HI-P197 68.5 63.8
71.5 >250 HI-P294 >250 >250 >250 >250 HI-P299 66.3
51.2 >250 >250
[0120]
11TABLE 5 Cytotoxic activity of fluoro-substituted dimethoxy
quinazolines on cancer cells. NALM-6 MOLT-3 U373 BT20 PC3 IC50 IC50
IC50 IC50 IC50 Compound (.mu.M) (.mu.M) (.mu.M) (.mu.M) (.mu.M)
HI-P144 28.1 .+-. 2.6 24.9 .+-. 3.7 49.5 .+-. 11.3 63.4 .+-. 5.5
>250 Hl-P214 >250 >250 >250 >250 >250 HI-P218
37.0 .+-. 5.8 33.2 .+-. 3.3 29.9 .+-. 7.3 37.62 .+-. 5.2 126.1 .+-.
5.8 HI-P219 22.3 .+-. 3.0 41.3 .+-. 4.4 83.6 .+-. 6.5 44.2 .+-.
10.9 58.3 .+-. 3.2 HI-P221 100.5 .+-. 4.8 98.73 .+-. 3.8 28.8 .+-.
12.7 30.67 .+-. 7.9 >250 HI-P223 39.5 .+-. 8.0 40.8 .+-. 15.1
32.1 .+-. 3.9 27.56 .+-. 8.6 >250 HI-P224 20.15 .+-. 8.1 23.3
.+-. 7.7 22.4 .+-. 5.9 58.33 .+-. 5.8 >250 HI-P228 57.3 .+-.
24.8 237.1 .+-. 4.8 >250 >250 >250 HI-P232 41.4 .+-. 6.9
43.6 .+-. 2.3 207.7 .+-. 18.1 70.54 .+-. 8.2 88.9 .+-. 17.2 HI-P264
47.0 .+-. 19.5 70.9 .+-. 17.3 53.3 .+-. 6.7 33.33 .+-. 7.5 >250
HI-P352 7.1 .+-. 1.8 21.8 .+-. 1.7 65.5 .+-. 11.2 50.3 .+-. 14.8
72.6 .+-. 2.5 HI-P353 6.1 .+-. 1.4 17.4 .+-. 1.5 14.5 .+-. 7.6 14.1
.+-. 3.3 64.9 .+-. 11.9 HI-P364 7.9 .+-. 1.9 25.3 .+-. 9.1 27.7
.+-. 1.2 40.1 .+-. 8.6 >250 HI-P365 86.5 .+-. 3.4 110.7 .+-. 7.5
>250 >250 >250 HI-P366 52.8 .+-. 14.0 137.2 .+-. 10.3 55.5
.+-. 13.2 61.7 .+-. 12.1 >250 HI-P369 >250 >250 >250
>250 >250 HI-P408 116.3 .+-. 17.8 228.5 .+-. 20.8 >250
>250 >250
[0121] The compounds were tested for activity against various
cancer cells. For example, NALM-6 cells were incubated with
HI-P144, HI-P214, HI-P221, HI-P224,.HI-P258, HI--P264, HI-P218,
HI-P223, HI-P228, HI-P366, HI-P367, HI-P219, HI-P352, HI-P353,
HI-P364 or HI-P365 for 24 hours in 96-well plates and cell survival
was determined by MTT assay. The data points represent the means
(.+-.SE) values from 3 independent experiments.
[0122] BT-20 breast cancer cells were incubated with HI-P144,
HI-214, HI-P221, HI-P224, HI-P258, HI-P264, HI-P218, HI-P223,
HI-P228, HI-P366, HI-P367, HI-P219, HI-P352, HI-P353, HI-P364, or
HI-P365 for 24 hours in 96-well plates and cell survival was
determined by MTT assay. The data points represent the mean
(.+-.SE) values from 3 independent experiments.
[0123] Apoptosis was induced by the compounds. The cells were
incubated with HI-P353 or HI-P219 for 24 hours, fixed,
permeabilized, and visualized for DNA degradation in a TUNEL assay
using digoxigenin-UTP-labeling kit. In the color photographs, red
fluorescence represents nuclei stained with propidium iodide. Green
or yellow (i.e., superimposed red plus green) represents the
apoptotic nuclei. Shown are control NALM-6 (FIG. 3A) and BT-20
(FIG. 3B) cells; HI-P353 treated NALM (FIG. 3D) and BT-20 cells
(FIG. 3E); and control (FIG. 3C) and HI-P219 treated PC3 cells
(FIG. 3F).
Example 8
Pharmacokinetic Studies
[0124] In pharmacokinetic studies, mice were injected either
intravenously (i.v.) via the tail vein or intraperitoneally (i.p.)
with a bolus dose of 300 .mu.g/mouse (.about.12.5 mg/kg=34
.mu.mols/kg) of HI-P131. Blood samples were obtained from the
ocular venous plexus by retroorbital venupuncture prior to and at 3
minutes, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes
and 1 hour, 2 hours, 4 hours, and 8 hours after administration of
HI-P131. All collected blood samples were heparinized and
centrifuged at 7,000.times.g for 10 minutes in a microcentrifuge to
obtain plasma. The plasma samples were stored at -20.degree. C.
until analysis. Aliquots of plasma were used for extraction and
HPLC analysis. Pharmacokinetic modeling and pharmacokinetic
parameter calculations were carried out using the pharmacokinetics
software, WinNonline Program, Version 1.1. (Scientific Consulting
Inc., Cary, N.C.). Concentration data were weighted by
I/concentration. An appropriate pharmacokinetic model was chosen on
the basis of lowest weighted squared residuals, lowest Schwartz
criterion (SC), lowest Akaike's Information Criterion (AIC) value,
lowest standard errors of the fitted parameters, and dispersion of
the residuals. The elimination half-life was estimated by linear
regression analysis of the terminal phase of the plasma
concentration profile. The area under the concentration time curve
(AUC) was calculated by the trapezoidal rule between first (0 h)
and last sampling time plus C/k, where C is the concentration of
last sampling and k is the elimination rate constant. Systemic
clearance (CL) was determined by dividing the dose by the AUC. The
apparent volume of distribution at steady-state was calculated
using the following equation,
V.sub.ss=Dose.multidot.AUMC/(AUC).sup.2. Bioavailability (F) was
estimated using the equation
F(%)=AUC.sub.ip.times.Dose.sub.iv/AUC.sub.iv.times.Dose.sub.ip.
[0125] HPLC Analysis of Plasma HI-P131 Levels:
[0126] A highly sensitive quantitative HPLC detection method (Chen
et al., 1998, J. Chromatography B (Biomedical sciences), in press)
was used to determine the pharmocokinetics of HI-P131. In brief,
the HPLC system consisted of a Hewlett Packard (HP) series 1100
equipped with an automated electronic degasser, a quaternary pump,
an autosampler, an automatic thermostatic column compartment, diode
array detector and a computer with a Chemstation software program
for data analysis. A 250.times.4 mm Lichrospher 100, RP-18 (5
.mu.m) analytical column and a 4.times.4 mm Lichrospher 100, RP-18
(5 .mu.m) guard column were obtained from Hewlett Packard Inc. (San
Fernando, Calif.). Acetonitrile/water containing 0.1% of
trifluoroacetic acid (TFA) and 0.1% triethylamine (TEA) (28:72,
v/v) was used as the mobile phase. The wavelength of detection was
set at 340 nm. Peak width, response time and slit were set at
>0.03 minutes, 0.5 seconds and 8 nm, respectively.
[0127] For determination of HI-P131 levels, 10 .mu.L of internal
standard HI-P154 (50 .mu.M) was added to a 100 .mu.L plasma sample.
For extraction, 7 ml chloroform was then added to the plasma
sample, and the mixture was vortexed thoroughly for 3 minutes.
Following centrifugation (300.times.g, 5 minutes), the aqueous
layer was frozen using acetone/dry ice and the organic phase was
transferred into a clean test tube. The chloroform extracts were
dried under a slow steady stream of nitrogen. The residue was
reconstituted in 100 .mu.L of methanol: water (9:1) and 50 .mu.L
aliquot of this solution was used for HPLC analysis. Under the
described chromatographic separation conditions, the retention
times for HI-P131 and HI-P154 were 5.1 minutes and 9.5 minutes,
respectively. At the retention time, HI-P131 and its internal
standard HI-P154 were eluted without any interference peaks from
the blank plasma.
[0128] HI-P131 was not toxic to mice at intraperitoneal single
bolus doses ranging from 0.5 mg/kg to 250 mg/kg. None of the 50
mice treated with HI-P1131 experienced side effects or died of
toxicity during the 30 day observation period. In particular, we
observed no hematologic side effects such as neutropenia,
lymphopenia, or anemia at the tested dose levels. No
histopathologic lesions were found in the organs of HI-P131 treated
mice that were selectively killed at 30 days and there was no bone
marrow hypoplasia or lymphoid cell depletion in spleen and lymph
nodes. Thus, the maximum tolerated dose (MTD) of HI-P131 was not
reached at 250 mg/kg. We next examined the pharmacokinetic features
of HI-P131 in mice. A two-compartment pharmacokinetic model was fit
to the pharmacokinetics data obtained following the intravenous
(i.v.) or intraperitoneal (i.p.) administration of a single
non-toxic 12.5 mg/kg bolus dose of HI-P131. The estimated maximum
plasma concentrations (Cma.) of HI-P131 were 85.6 .mu.M after i.v.
administration.
Example 9
Antitumor Activity of Quinazolines In Vivo
[0129] To test the anti-tumor activity of quinazolines in vivo,
cancer cells were implanted and grown in mice in the presence of
quinazoline.
[0130] Inhibition of Breast Cancer Cells
[0131] The left hind legs of CB.17 SCID mice were inoculated
subcutaneously with 0.75.times.10.sup.6 MDA-MB-231 human breast
cancer cells in 0.1 ml PBS. Twenty-four hours after inoculation,
the mice were treated with HI-P353 (10 mg/kg/day x 5 days/week,
N=7), or HI-P364 (10 mg/kg/day.times.5 days/week, N=8), or vehicle
(50% DMSO in PBS, N=7) for four weeks. The mice were monitored
daily for health status and tumor growth. Measurements were taken
on the tumors 3 times a week using a Vernier caliper. Tumor volumes
were calculated using the following formula:
(width).sup.2.times.(length/2). Comparisons of the outcomes of the
three groups were done using the log-rank test.
[0132] The data are shown in FIG. 10A, and demonstrate that
treatment of animals with the quinazolines of the invention
(HI-P353 and HI-P364) inhibited the growth of breast cancer cell
tumors as compared with untreated controls.
[0133] Inhibition of Brain Tumor Cells
[0134] An analogous experiment was done implanting brain tumor
cells into mice. The right hind legs of CB.17 SCID mice were
inoculated subcutaneously with 1.times.10.sup.6 of U373 human
glioblastoma cells in 0.1 ml of PBS. Twenty four hours after
inoculation, mice were treated with HI-P353 (10 mg/kg/day.times.5
days/week, N=7), or HI-P364 (10 mg/kg/day.times.5 days/week, N=8),
or vehicle (50% DMSO in PBS, N=7) for four weeks. Mice were
monitored daily for health status and tumor growth. Tumors were
measured 3 times a week using a Vernier caliper. Tumor volumes were
calculated using the following formula:
(width).sup.2.times.(length/2). Comparisons of outcome between
groups were done using the log-rank test.
[0135] The data are shown in FIG. 10B and demonstrate that
treatment of animals with the quinazolines of the invention
(HI-P353 and HI-P364) inhibited the growth of brain tumors as
compared with untreated controls.
[0136] Inhibition of Intracranial Brain Tumors
[0137] The anti-tumor activity of the quinazolines of the invention
was also studied with intracranial tumors. Nude mice were first
anesthetized with Avertin. Under aseptic conditions in a laminar
flow hood, a small hole was drilled at 2 mm to the right of the
midline and 2 mm posterior to the bregma. An amount of
4.times.10.sup.5 U87 glioblastoma cells in 10 .mu.L of PBS were
intracranially implanted using a Hamilton syringe into the right
cerebral hemisphere of mice and a stereotaxic apparatus according
to the method described in Huang, H. J. S. et al., J. Biol. Chem.
272:2927-2935, 1997.
[0138] Twenty-four hours after inoculation, mice were treated with
1H--P353 (20 mg/kg/day.times.10 days, n=10), HI-P364 (20
mg/kg/day.times.10 days, n=10), or vehicle (50% DMSO in PBS, n=10).
The mice were monitored twice daily for health status.
[0139] FIG. 10C shows the survival rate of mice inflicted with
intracranial tumors. Treatment of mice with quinazolines (HI-P353
and HI-P364) resulted in prolonged survival as compared with mice
treated with vehicle alone.
[0140] All publications, patents, and patent documents described
herein are incorporated by reference as if fully set forth. The
invention described herein may be modified to include alternative
embodiments. All such obvious alternatives are within the spirit
and scope of the invention, as claimed below.
* * * * *