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Publication numberUS20010018539 A1
Publication typeApplication
Application numberUS 09/797,965
Publication dateAug 30, 2001
Filing dateMar 5, 2001
Priority dateApr 14, 1999
Also published asUS6369246, WO2000061536A2, WO2000061536A3
Publication number09797965, 797965, US 2001/0018539 A1, US 2001/018539 A1, US 20010018539 A1, US 20010018539A1, US 2001018539 A1, US 2001018539A1, US-A1-20010018539, US-A1-2001018539, US2001/0018539A1, US2001/018539A1, US20010018539 A1, US20010018539A1, US2001018539 A1, US2001018539A1
InventorsHsu-Shan Huang, Kung-Yuan Lee, Chang-Hsin Shi, Hsien-Chin Hsu
Original AssigneeHsu-Shan Huang, Kung-Yuan Lee, Chang-Hsin Shi, Hsien-Chin Hsu
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Antiallergens, antiinflammatory agents, antitumor agents
US 20010018539 A1
Abstract
The invention relates to novel anthracene compounds useful in the treatment of allergic, inflammatory conditions, tumor conditions and therapeutic compositions containing such compounds. The compounds of the present invention are 9-acyloxy-substituted 1,5-dichloroanthracene or analogs thereof. According to the practice of the invention, there are provided 9-acyloxy substituted 1,5-dichloroanthracene compounds according to formula III:
wherein R represents a straight or branched chain alkyl group having 1 to 6 carbons which may be substituted with one or more groups of R1, or R represents phenyl or benzyl which may be substituted with one or two groups of R2; wherein R1 is selected from the group consisting of halogen, —NO2, —OCH3, OCH2CH3, and —OCH2CH2CH3; and wherein R2 is selected from the group consisting of a straight or branched chain alkyl group having 1 to 4 carbons, halogen, —NO2, —OCH3, —OCH2CH3, and —OCH2CH2CH3.
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Claims(31)
What is clamed is:
1. A compound according to Formula III,
wherein R represents a straight or branched chain alkyl group having 1 to 6 carbons which may be substituted with one or more groups R1, or R represents a phenyl or benzyl group which may be substituted with one or two groups R2;
wherein R1 is selected from the group consisting of halogen, —NO2, —OCH3, OCH2CH3, and —OCH2CH2CH3; and
wherein R2 is selected from the group consisting of a straight or branched chain alkyl group having 1 to 4 carbons, halogen, —NO2, —OCH3, —OCH2CH3, and —OCH2CH2CH3.
2. The compound according to
claim 1
, wherein R represents a phenyl or benzyl group having at least one substituent selected from the group consisting of methyl, NO2, OCH3 and Cl.
3. The compound according to
claim 1
, wherein R represents a substituted phenyl group selected from the group consisting of 2-CH3C6H4, 3-CH3C6H4, 4-CH3C6H4, 3-ClC6H4, 4-ClC6H4, 3-NO2C6H4, 4-NO2C6H4, 4-Cl-2-CH3OC6H3, and 2,4-Cl2C6H3.
4. The compound according to
claim 1
, wherein R is selected from the group consisting of CH2C6H5 and CH2CH2C6H5.
5. The compound according to
claim 1
, wherein R represents a alkyl group selected from the group consisting of CH3, CH2Br, CH2Cl, CH2CH3, CH(CH3)2, CH(CH3)Cl, CHCl2, (CH2)2CH3, (CH2)3Br, (CH2)Cl, and (CH2)4CH3.
6. A method for inhibiting or treating an allergic or inflammatory condition, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 1
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
7. A method for anti-cancer treatment, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 1
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
8. A method for treating abnormal proliferation, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 1
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
9. A method for enhancing an anti-oxidation affect, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 1
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
10. An anti-cancer drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 1
.
11. An anti-inflammatory drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 1
.
12. An antioxidant, comprising, as an active ingredient, the pharmaceutical compound of
claim 1
.
13. An anti-dermatitis cancer drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 1
.
14. A method for synthesis of 9-substituted-1,2-dichloroanthrecene derivatives, comprising reacting 1,5-dichloroanthrone with an acyl chloride under weakly basic conditions to give a 9-acyloxy-1,5-dichloroanthrancene according to Formula III
wherein R represents a straight or branched chain alkyl group having 1 to 6 carbons which may be substituted with one or more groups R1, or R represents a phenyl or benzyl group which may be substituted with one or two groups R2;
wherein R1 is selected from the group consisting of halogen, —NO2, —OCH3, OCH2CH3, and —OCH2CH2CH3; and
wherein R2 is selected from the group consisting of a straight or branched chain alkyl group having 1 to 4 carbons, halogen, —NO2, —OCH3, —OCH2CH3, and —OCH2CH2CH3.
15. The method of
claim 14
, wherein the weakly basic condition is accomplished by the addition of pyridine.
16. A pharmaceutical compound which is synthesized by reacting 1,5-dichloroanthrone with an acyl chloride of the form RC(O)Cl under a weakly basic condition; wherein R represents a straight or branched chain alkyl group having 1 to 6 carbons which may be substituted with one or more groups R1, or R represents phenyl or benzyl which may be substituted with one or two groups R2;
wherein R1 is a group selected from halogen, —NO2, —OCH3, —OCH2CH3, and —OCH2CH2CH3; and
wherein R2 is a group selected from a straight or branched chain alkyl group having 1 to 4 carbons, halogen, —NO2, —OCH3, —OCH2CH3, and —OCH2CH2CH3.
17. The pharmaceutical compound according to
claim 16
, wherein said weakly basic condition is accomplished by the addition of pyridine.
18. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is benzoyl chloride or a substituted benzoyl chloride which is selected from the group consisting of 2-methylbenzoyl chloride, 3-methylbenzoyl chloride, 4-methylbenzoyl chloride, 3-chlorobenzoyl chloride, 4-chlorobenzoyl chloride, 3-nitrobenzoyl chloride, 4-nitrobenzoyl chloride, 4-chloro-2-methoxybenzoyl chloride and 2,4-dichlorobenzoyl chloride.
19. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is phenylacetyl chloride or a substituted phenylacetyl chloride having a substituent selected from the group consisting of methyl, nitro, methoxy and chloro.
20. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is acetyl chloride, bromoacetyl chloride, chloroacetyl chloride, propanoyl chloride, isobutyryl chloride, 2-chloropropanoyl chloride, dichloroacetyl chloride, butyryl chloride, 4-bromobutyrlyl chloride, 4-chlorobutyrlyl chloride, 3-phenylpropanoyl chloride or hexanoyl chloride.
21. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is an acetyl chloride having a 1H-NMR characterized by signals at δ (ppm relative to tetramethyl silane) of approximately 8.79 (s), 8.01 (d), 7.90 (d), 7.62-7.58 (m), 7.43-7.34 (m), 2.60 (s), wherein s, d and m designate signals comprising a singlet, a doublet and a multiplet, respectively.
22. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is benzoyl chloride and having a 1H-NMR characterized by signals at δ (ppm relative to tetramethyl silane) of approximately 8.82 (s), 8.40 (d), 8.01 (d), 7.91 (d), 7.71 (t), 7.61-7.58 (m), 7.56-7.52 (d), 7.37-7.33 (m), wherein s, d, t and m designate signals comprising a singlet, a doublet, a triplet, and a multiplet, respectively.
23. The pharmaceutical compound according to
claim 16
, wherein the acyl chloride is 3-methylbenzoyl chloride and having a 1H-NMR characterized by signals at δ (ppm relative to tetramethyl silane) of approximately 8.74 (s), 8.15 (d), 7.93 (d), 7.85 (dd), 7.53-7.48 (m), 7.46-7.39 (m), 7.29-7.25 (m), 2.42 (s) wherein s, d, dd and m designate signals comprising a singlet, a doublet, a doublet of doublets, and a multiplet, respectively.
24. A method for inhibiting or treating an allergic or inflammatory condition, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 16
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
25. A method for anti-cancer treatment, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 16
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
26. A method for treating abnormal proliferation, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 16
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
27. A method for enhancing an anti-oxidation affect, comprising administering a therapeutically effective amount of a pharmaceutical compound according to
claim 16
or a pharmaceutically acceptable salt of said compound and optionally a pharmaceutical carrier to a patient in need of such treatment.
28. An anti-cancer drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 16
.
29. An anti-inflammatory drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 16
.
30. An antioxidant, comprising, as an active ingredient, the pharmaceutical compound of
claim 16
.
31. An anti-dermatitis cancer drug, comprising, as an active ingredient, the pharmaceutical compound of
claim 16
.
Description
RELATED APPLICATION

[0001] This Application is a continuation-in-part of U.S. patent application Ser. No. 09/290,865, filed on Apr. 14, 1999, which is herein incorporated by reference.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] This invention relates to novel anthracene compounds useful in the treatment of allergic, inflammatory conditions, tumor conditions and therapeutic compositions containing such compounds. The invention relates also to the therapeutic compositions effective at low dose with low irritancy. These anthracene compounds possess antitumor, antiproliferative, antipsoriatic, antiinflammatory, or antioxidant activity.

[0004] 2. Description of the Prior Art

[0005] The discovery of the antitumor activity of 1,4-bis[(aminoalkyl)amino]anthracene-9, 10-diones such as ametantrone (1) and mitoxantrone (2) (FIG. 1) (Zee-Cheng, R. K. V. et al., J. Med. Chem., 21, 291-294 (1978); Zee-Cheng, R. K. V. et al., J. Pharm. Sci., 71, 708-709 (1982); Murdock, K. C. et al., J. Med. Chem., 22, 1024-1030 (1979)) has led to numerous physicochemical and pharmacological studies on the tumoricidal mechanisms of these chemotypes. Krapcho, A. P. et al., J. Med. Chem., 341, 2373-2380 (1991); Morier-Teissier, E. et al., J. Med. Chem., 36, 2084-2090 (1993). Additional references disclose 1,4- and 2,6-disubstituted or regioisomeric amidoanthracene-9,10-dione derivatives as inhibitors of human telomerase. Perry, P. J. et al., J. Med. Chem., 41, 3253-3260 (1998) and Perry, P. J. et al., J. Med. Chem., 41, 4873-4884 (1998).

[0006] Although the active mechanism of the antitumor activity of the anthracene-9,10-diones such as ametantrone (1) and mitoxantrone (2) is probably multimodal in nature, a number of studies have indicated that an intercalative interaction with DNA may be a major cellular event. Denny, W. A., Anti-Cancer Drug Design, 4, 241-263 (1989). Antitumor quinones represent the second largest class of clinically approved anticancer agents in the U.S.A., second only to the chloroethyl alkylating agents. They have been selected from the large number of naturally occurring quinones (Moore, H. W et al., Drugs Expl. Clin. Res., 12, 475-494, (1986)) and from synthetic quinones. Bruce, J. M. ed., Benzoquinones and Related Compounds, Vol. 3, Part 4, 1-306, (1974). The planar tricyclic system is known to intercalate into DNA base pairs and interfere in the transcription and replication processes of the cell. Johnson, R. K. et al., Cancer Treat. Rep., 63, 425-439 (1979); Lown, J. W. et al., Biochemisty, 24, 4028-4035 (1985). The DNA binding affinity (quantified as a binding affinity constant) and the dissociation rate constant for the DNA-ligand complex have been evaluated. Drug-DNA binding constants for ametantrone (1), mitoxantrone (2) and related congeners with calf thymus DNA show a large sensitivity to the position and number of the OH substitutions and the nature of the charged side chain. Denny, W A., Anti-Cancer Drug Design, 4, 241-263 (1989).

[0007] Normal human cells undergo a finite number of cell divisions and ultimately enter a nondividing state called replicative senescence. During successive rounds of cell division, the end-replication problem results in telomere shortening and ultimately senescence. As such, the loss of telomeric repeats after each round of cell division has been likened to a “biological clock” limiting the proliferative life span of normal somatic cells. Harley, C. B. et al., Nature, 345, 458-460 (1990). Consequently, telomerase has been proposed as a potentially highly selective target for the development of a novel class of antiproliferative agents. However, in order for a therapeutic treatment to be effective, both the inflammatory and hyperproliferative aspects of the condition must be addressed. Substantial evidence suggests that free radicals and active oxygen species play a key role in both the therapeutic activity and side effects of anthracenone derivatives.

[0008] Anthraquinone-based compounds currently occupy a prominent position in cancer chemotherapy, with the naturally occurring aminoglycoside anthracycline doxorubicin and the aminoanthraquinone mitoxantrone both being in clinical use. These 1,5-dichloro-9(10H)-anthracenone compounds contain alkylacyl or arylacyl moieties at the C-9 position resulting in enhanced antiproliferative activity of the compounds. These blocked compounds may be further modified by introducing the phenolic form of the arylacyl or alkylacyl substituent.

[0009] As noted above, cancer is typically characterized by hyperproliferative component. There is thus a continuing need for effective compounds that address these aspects of cancer disease.

SUMMARY OF THE INVENTION

[0010] The present invention is directed to novel 9-substituted-1,5-dichloroanthracene compounds and salts thereof having therapeutic utility with respect to allergic or inflammatory or tumor conditions. In particular, many of the improved anthracene compounds provided for according to the invention are effective at low concentrations for the treatment of patients suffering from allergic or inflammatory or tumor conditions. Because these compounds may be administered at low concentrations, the undesirable allergic or inflammatory effects caused in whole or in part by free radicals or active oxygen species that are generated by anthracenone compounds are substantially eliminated.

[0011] Accordingly, in one embodiment of the invention, there is provided an anthracene compound according to Formula III below, said compound containing a substituent R, wherein R represents a branched or straight chain alkyl group having from 1 to 4 carbon atoms, said alkyl group being substituted with at least one substituent selected from carboxyl, carboxyl ester, hydroxy, phenyl, benzyl, substituted benzyl and substituted phenyl groups.

[0012] In a preferred embodiment of the invention, R represents a substituted phenyl group having at least one substituent selected from methyl, halogen and nitro groups. In another preferred embodiment, R represents a straight or branched chain alkyl group having 1 to 4 carbon atoms, which may contain a substituent selected from acyl and phenyl groups. Additionally, there are provided compounds which are functional analogs of the compounds of Formula III.

[0013] As aforementioned, therapeutic compositions of the invention are effective at dosages that substantially eliminate the adverse inflammatory or irritancy effects associated with the use of anthracenone and related compounds. Accordingly, there is provided a therapeutic composition comprising a therapeutically effective amount of at least one compound of the invention and a pharmaceutically acceptable carrier. These compounds of the invention have antiproliferative effects and antineoplastic effects.

[0014] Further additional representative and preferred aspects of the invention are described below according to the following detailed description of the invention.

DESCRIPTION OF THE DRAWINGS

[0015]FIG. 1 Shows the structure of prior art anthraquinonic derivatives.

[0016]FIG. 2 is an outline of the synthesis of the 9-substituted-1,5-dichloroanthracene compounds.

[0017]FIG. 3: Perspective view of the molecular structure of compound (3o).

DETAILED DESCRIPTION OF THE INVENTION

[0018] As set forth above, the compounds of the invention are 9-acyloxy-1,5-dichloroanthracene analogs. According to of the invention, there are provided 9-substituted-1,5-dichloroanthracene compounds according to Formula III.

[0019] wherein R represents a straight or branched chain alkyl group having 1 to 6 carbon atoms, phenyl or benzyl, wherein the alkyl group may be substituted with one or more groups R1 and the phenyl or benzyl group may be substituted with one or two groups R2. R1 is a group selected from halogen, NO2, CH3O, CH3CH2O, and CH3CH2CH2O. R2 is a group selected from a straight or branched chain alkyl group having 1 to 4 carbon atoms, halogen, NO2, CH3O, CH3, CH2O, CH3CH2CH2O.

[0020] In preferred compounds according to the invention, R represents a straight or branched chain alkyl group having 1 to 4 carbon atoms which may be substituted with one or more groups R1, selected from Cl, NO2, CH3O. In other preferred embodiments, R is a phenyl or benzyl group having one or two substituents R2, selected from a straight or branched chain alkyl group having 1 to 4 carbon atoms, Cl, NO2, CH3O. Suitable compounds of the invention described in Table 1 (infra).

[0021] In the course of synthesis of the 9-substituted-1,5-dichloroanthracenes, it was found that the molecule reacted to the acylating agents in an unusual manner. Introduction of the 9-acyloxy functionality onto the anthracene nucleus (compounds 3a-w) was achieved by reaction of the appropriate acyl chlorides with 1,5-dichloroanthrone under weakly basic conditions, e.g. pyridine, where ester formation takes place at the C-9 oxygen via the enol tautomer (FIG. 2).

[0022] For the pharmaceutical compositions according to the invention, salts of 9-substituted-1,5-dichloroanthracene compounds are in particular salts with the pharmaceutically acceptable base. Excipients such as magnesium stearate, corn starch, starch, lactose, sodium hydroxymethylcellulose, ethanol, glycerol etc. may be added in the preparation of pharmaceutical compositions containing 1,5-dichloro-9-acyloxyanthracene derivatives of the present invention. The pharmaceutical compositions of the invention may be in an injectable form or formulated into tablet, pill or other solid preparation forms. The pH value for injectable forms may be adjusted with phosphate buffer. Generally, dosage used for injectable forms is 25-100 mg. For solid preparations, an effective dosage is 3-500 mg, administered 2 to 3 times a day.

Clinical Indications Subject to Treatment

[0023] The following conditions are selected for description herein as being representative of inflammatory, allergic, antioxidant, or neoplastic conditions that are suitable for treatment according to the practice of the invention. Each of these conditions involves intimation hyperproliferation and/or generation of free radicals and active oxygen species.

[0024] Neoplastic Conditions

[0025] The therapeutic compositions of the invention may be used in the treatment of a wide variety of cancers such as carcinomas, sarcomas, melanomas and lymphomas, which may affect a wide variety of organs, including, for example, the lungs, mammary tissue, prostate gland, small or large intestine, liver, heart, skin, pancreas and brain. The therapeutic compositions may be administered by injection (intravenously, intralesionally, peritoneally, subcutaneously), or by topical application and the like as would be suggested according to the routine practice of the art.

[0026] Psoriasis and Contact Dermatitis

[0027] Psoriasis is a widespread, chronic, inflammatory and scaling skin disease. Contact dermatitis, in contrast, is a short term allergic condition characterized by scaling skin. Both psoriasis and contact dermatitis are characterized by increased epidermal cell proliferation at the affected site or sites, i.e. lesions. Muller, K., et al., J. Med. Chem., 39, 3132-3138 (1996).

[0028] Arthritic Disease

[0029] Rheumatoid arthritis is a chronic inflammatory disease, primarily of the joints, that may result in permanent loss of joint function. Irreversible loss of joint function is attributed to severe degradation of collagen, bone, ligament and tendon. Associated chronic intimation results, in part, from immune response at the affected joint, although the exact nature of the triggering antigens is unknown. The immune response may be autoimmune in origin. Mullins, D. E. and Rohrlich, S. T., Biochemica et Biophysica Acta, 695, 177-214 (1983). The etiology has been described is in detail. (pp. 192-193.) Briefly, there is a progressive loss of cartilage (a connective tissue) caused by invading cells. Both collagen and proteoglycan components of the cartilage are degraded by enzymes released at the affected site.

[0030] Therapeutic Compositions and Administration Thereof

[0031] The amount of 9-substituted-1,5-dichloroanthracene (or salt thereof) administered for the prevention or inhibition of an inflammatory or allergic condition, for antiproliferative or antineoplastic effect, can be determined readily for any particular patient according to recognized procedures. Additional information useful in the selection of therapeutic compositions is provided as follows. For use in the treatment of inflammatory or degenerative conditions, as those term are recognized in the art, the therapeutic compositions may be administered, for example, by injection at the affected site, by aerosol inhalation (as in the case of emphysema or pneumonia), or by topical application or transdermal absorption as would also be suggested according to the routine practice of the art.

[0032] As described above, the 9-substituted-1,5-dichloroanthracene (or salt thereof) may be incorporated into a pharmaceutically acceptable carrier or carriers for application (directly or indirectly) to the affected area. The nature of the carrier may vary widely and will depend on the intended location of application and other factors well known in the art. Such carriers of anthralin or anthracenone compounds are well known in the art. See, for example, Kammerau, B. et al., J. Investigative Dermatology, 64, 145-149 (1975).

Preparation of the Compounds of the Invention

[0033]FIG. 2 is an outline of a synthesis of the 9-substituted-1,5-dichloroanthracene compounds (Formula III) according to the invention. As shown in FIG. 2, reduction of 1,5-dichloroanthraquinone (1) with SnCl2 in boiling HCl and acetic acid proceeds with concomitant ether cleavage and leads to the corresponding 1,5-dichloro-9(10H)-anthracenone (II). In the course of synthesis of the 9-substituted-1,5-dichloroanthracene, it was found that the molecule reacted to the acylating agents in an unusual manner. Introduction of the 9-acyloxy functionality onto the anthracene nucleus (compounds 3a-w) was achieved by reaction of the appropriate acyl chlorides with 1,5-dichloro-9(10H)-anthrone under weakly basic conditions (pyridine), resulting in ester formation at the C-9 position via the enol tautomer (FIG. 2). When the anthrone was allowed to react with acyl chlorides in CH2Cl2 in the presence of a catalytic amount of pyridine, the reaction time is reduced compared to the noncatalyzed reaction. Specific methods for the preparation of several compounds according to the present invention are described below in Example 1. The structure of each of the synthesized compounds is confirmed by 1H-NMR spectrometry, mass spectrometry and elemental analysis as shown in Example 2. Procedures adapted from the descriptions and the following non-limiting examples will allow one skilled in the art to prepared similar compounds of the invention.

EXAMPLES

[0034] The following non-limiting examples are representative of the practice of the invention.

Example 1 Methods of Synthesis

[0035] The novel 9-substituted-1,5-dichloroanthracene compounds described in Table 1 were produced as follows.

[0036] 1,5-dichloroanthraquinone (1) was reduced with SnCl2 in boiling HCl and acetic acid with ether cleavage to give the corresponding 1,5 dichloro-9(10H)-anthracenone (II). To a solution of 1,5-dichloro-9(10H)anthracenone (1 mmol) and 0. 1 mL of pyridine in dry CH2Cl2 (20 mL) was added dropwise a solution of an appropriate acyl chloride (3 mmol) in dry CH2Cl2 (10 mL) under N2. The reaction mixture was stirred at room temperature or refluxed for several hours. The solvent was removed and the residue purified by recrystallization and chromatography. This procedure was used to synthesize each of the compounds in Table 1.

Example 2 Structural Confirmation

[0037] All temperatures are reported in degrees centigrade. Melting points were determined with a Büchi 530 melting point apparatus and are uncorrected. Chromatography refers to column chromatography using silica gel (E. Merck, 70-230 mesh). 1H-NMR spectra were recorded with a Varian GEMR-H-300 (300 MHz); δ values are in ppm relative to a tetramethylsilane internal standard. Fourier-transform IR spectra (KBr) were recorded on a Perkin-Elmer 983G spectrometer. Mass spectra (EI, 70 eV, unless otherwise stated) were obtained on a Finnigan MAT TSQ-46 and Finnigan MAT TSQ-700. UV spectra were recorded on a Shimadzu UV-160.

[0038] (1) 1,5-Dichloro-9-acetyloxy-anthracene (3a)

[0039] The compound was synthesized as in Example 1 and analyzed by 1H-NMR (500 MHz CDCl3), δ (ppm): 8.79 (H, s, H-10), 8.01 (H, d, J=8.4 Hz, H-2), 7.90 (H, d, J=8.8,H6), 7.62-7.58 (2H, m, H-4,8), 7.43-7.34 (2H, m, H-3,7), 2.60 (3H, s, COCH3); 13C-NMR: (75 MHz, CDCl3), δ (ppm): 170.58, 142.00, 134.51, 132.31, 130.54, 129.87, 129.61, 128.37, 126.97, 126.93, 126.80, 125.97, 123.81, 121.79, 121.67, 22. 1; MS m/z 304 (7), 262 (100); Anal. (Cl16H10O2Cl2); C, H.

[0040] (2) 1,5-Dichloro-9-benzoyloxy-anthracene (3L)

[0041] The compound was synthesized as in Example 1 and analyzed by 1H-NMR: (500 MHz, CDCl3), δ (ppm): 8.82 (H, s, H-10), 8.40 (2H, d, J=8.1 Hz, H-2′,6′), 8.01 (2H, d, J=8.4 Hz, H-2), 7.91 (H, d, J=8.8 Hz, H-6), 7.71 (H, t, J 7.5 Hz, H-4′), 7.61-7.58 (2H, m, H-4,8), 7.56-7.52 (2H, d, J=7.2 Hz, H-3′,5′), 7.37-7.33 (2H, m, H-3,7); 13C-NMR: (75 MHz, CDCl3), δ(ppm): 166.62, 142.41, 134.49, 132.64, 131.28, 130.43, 130.11, 129.98, 129.53, 129.40, 128.65, 127.24, 126.98, 126.00, 125.84, 122.45, 121.81; MS m/z 367 (10), 262 (7), 246 (4), 227 (5), 105 (100); Anal. (C21H12O2Cl2); C, H.

[0042] (3) 1,5-Dichloro-9-(m-toluyloxy)-anthracene (3n)

[0043] The compound was synthesized as in Example 1 and analyzed by 1H-NMR: (500 MHz, CDCl3), δ (ppm): 8.74 (H, s, H-10), 8.15 (H, d, J=7.4 Hz, H-2′,6′), 7.93 (H, d, J=8.8,H-2), 7.85 (H, dd, J=8.8, 0.9 Hz, H-6), 7.53-7.48 (2H, m, H-4,8), 7.46-7.39 (2H, m, H-4′,5′), 7.29-7.25 (2H, m, H-3,7), 2.42 (3H, S, COCH3); 13C-NMR: (75 MHz, CDCl3), 6 (ppm): 166.41, 142.98, 139.28, 135.31, 134.55, 132.58, 131.83, 130.40, 129.94, 129.90, 129.52, 129.31, 128.72, 128.55, 127.19, 126.94, 126.00, 123.79, 122.38,121.86,21.97; MS m/z 381 (13), 119 (100); Anal. (C22H14O2Cl2); C, H.

[0044] (4) 1,5-Dichloro-9-(p-toluoxy)-anthracene (3o)

[0045] The compound was synthesized as in Example I and analyzed by 1H-NMR: (500 MHz, CDCl3), δ (ppm): 8.81 (H, s, H-10), 8.28 (2H, d, J=8.1 Hz, H-2′,6′), 8.01 (H, d, J=8.4 Hz, H-4), 7.91 (H, d, J=8.8,H-5′), 7.36-7.32 (2H, m, H-3,7), 2.58 (3H, s, CH3); 13C-NMR: (75 MHz, CDCl3), δ (ppm): 166.28, 145.43, 143.43, 134.56, 132.56, 131.43, 130.38, 130.13, 129.91, 129.51, 128.76, 127.28, 127.24, 126.97, 126.91, 125.99, 123.74, 122.49, 121.89, 22.44; MS m/z 380 (4), 119 (100); Anal. (C22H14O2Cl2); C, H.

[0046] The perspective view of the molecular structure of compound (3o) is shown in FIG. 3.

Example 3 Growth Inhibition Assay

[0047] Growth inhibition was measured in three human carcinoma cell lines (GBK KB and CHO) using a previously described in vivo assay. Hwang, J.-M. et al., Chin. Med. J (Taipei), 51, 166-175 (1993). Human oral epidermoid carcinoma cells (KB cell line), human cervical carcinoma cells of ME 180 (GBM8401) and Chinese hamster ovary (CHO) cells grown in plateau phase were used in all experiments. Each cell line was further divided into control and experimental groups, respectively.

[0048] Stock solutions of the test compounds were prepared in DMSO and diluted with DMEM to give a final concentration of DMSO of 0.2%. Controls were performed with DMSO or with medium alone. The medium was removed and each well was rinsed with 100 μL PBS 48 hours after addition of the test compound to the culture. The cells were then incubated with sterile 0.5% trypsin, 0.2% EDTA in PBS for 20 minutes at 37° C. The detached cells from each well were suspended in DMEM and dispersed into single cells by gentle pipetting through an Eppendorf pipette and cell growth was determined directly by counting the cells in a Neubauer counting chamber using phase contrast microscopy. Inhibition was calculated by comparison of the mean values of the test compound (N=3) with the control (N=6-8) activity: (1-test compound/control)×100. Inhibition was statistically significant compared to that of control (Student's West; P=0.05). IC50 values (concentration required to inhibit cell growth by 50%) were determined for each agent which was derived by interpolation of a log inhibitor concentration versus response plot using four or more different concentrations of the compound spanning the 50% inhibition point.

[0049] Several compounds of the invention had an antiproliferative IC50 value of less than 1.1 μM for GBM cell line. In particular, compounds 3g, 3j and A had IC50 values of 1.4, 1.1 and 1.2 μM respectively. In addition, each of compounds 3j and 3v showed an 1C50 value of 11.0 μM and 12.6 μM in the KB assay. The results of this assay are provided in Table 1.

Example 4 Taq Polymerase Assay

[0050] Prior to the evaluation of compounds in the PCR-based telomerase assay, the agents were tested for their ability to inhibit Taq polymerase in order to address the selectivity of polymerase/telomerase inhibition. Compounds were included at both 10 and 50 μM final concentrations in a PCR 50 μL master mix comprising 10 ng of pCI-neo mammalian expression vector (Promega, Southampton, U.K.), forward (CGAGTTCCGCGT-TACATAAC) and reverse (GTCTGCTCGAAGCATTAACC) primers (200 nmol), reaction buffer (75 mM Tris-HCl, pH 8.81, 20 mM (N4H4)2SO4, 0.01% v/v Tween 20), 2.5 mM MgCl2, 200 μM of each deoxynucleotide triphosphate, and thermostable DNA polymerase (“red hot”, Advanced Biotechnologies, 1.25 units). A reaction mix containing water and no drug was used as a positive control, producing a product of approximately 1 kb. Amplification (30 cycles of 94° C. for 1 min, 55° C. for 1 min, and 72° C. for 2.5 min) were performed using a thermal cycler (Hybaid, U.K.). PCR products were then separated by electrophoresis on a 2% w/w agarose gel and visualized using ethidium bromide. The results of this assay are provided in Table 2.

Example 5 Lipid Peroxidation Assay

[0051] Rat brain homogenate was prepared from the brains of freshly killed Wistar rats and its peroxidation. In the presence of iron ions was measured by the thiobarbituric acid (TBA). Teng, C. M. et al., Eur. J. Pharmacol., 303, 129-139 (1996). Tetramethoxypropane was used as a standard, and the results were expressed as nanomoles of malondialdehyde equivalents per milligram of protein of rat brain homogenates.

[0052] In brief, whole brain tissue, excluding the cerebellum, was washed and homogenized in 10 volumes of ice-cold Krebs buffer (10 mM N-2 hydroxyethyl-piperazine-N′-2-ethanesulfonic acid (Hepes), 10 mM glucose, 140 mM NaCl, 3.6 mM KCl, 1.5 mM CaCl2, 1.4 mM KH2PO4, 0.7 mM MgSO4, pH 7.4) using a glass Dounce homogenizer. The homogenate was centrifuged at low speed (1000×g) for 10 min, and the resulting supernatant (adjusted to 2 mg/mL) was used immediately in lipid. peroxidation assays.

[0053] The reaction mixture with test compounds or vehicle was incubated for 10 min, then stimulated by addition of ferrous ion (200 μM freshly prepared), and maintained at 37° C. for 30 min. The reactions were terminated by adding 10 μM of ice-cold trichloroacetic acid solution (4% (w/v) in 0.3 N HCl) and 200 μM, of thiobarbituric acid-reactive substance reagent (0.5% (w/v) thiobarbituric acid in 50% (v/v) acetic acid). After boiling for 15 min, the samples were cooled and extracted with n-butanol. The extent of lipid peroxidation was estimated as thiobarbituric acid-reactive substances and was read at 532 nm in a spectrophotometer (Shimadzu UV-160). The results of this assay are provided in Table 3.

[0054] The contents of all patents, patent applications, published articles, books, reference manuals and abstracts cited herein are hereby incorporated by reference in their entirety to more fully describe the state of the art to which the invention pertains.

[0055] As various changes can be made in the above-described subject matter without departing from the scope of the invention, it is intended that all subject matter contained in the above description, shown in the accompanying drawing, or defined in the appended claims, be interpreted as descriptive, illustrative, and non-limiting. Modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described.

TABLE 1
In vitro cytotoxicity data for novel 1,5-dichioroanthracene compounds of the invention
Comp'd mp IC50(μM)a
No. R group Formula MW (° C.) GBM KB CHO
3a CH3 C16H10O2Cl2 305.16 164-166 48.1 472.8 114.5
3b CH2Br C16H9O2Cl2Br 384.06 180-181 19.2 136.0 1012.8
3c CH2Cl C16H9O2Cl3 339.60 181-182 105.6 527.5 1.2
3d CH2CH3 C17H12O2Cl2 319.18 134-135 17.2 470.8 1157.6
3e CH(CH3)2 C18H14O2Cl2 332.17 116-118 195.5 430.8 1314.2
3f CH(CH3)Cl C17H11O2Cl3 353.63 120-122 125.8 374.1 954.9
3g CHCl2 C16H8O2Cl4 374.05 138-140 1.4 527.8 1277.9
3h (CH2)2CH C18H14O2Cl2 333.21 132-134 572.2 544.3 0.1
3i (CH2)3Br C18H13O2BrCl2 412.11 118-120 205.2 457.3 109.9
3j (CH2)3Cl C18H13O2Cl3 367.65 130-132 1.1 11.0 1171.3
3k (CH2)4CH3 C20H18O2Cl2 361.26 120-121 1.2 316.8 55.1
3L C6H5 C21H12O2Cl2 367.23 166-168 226.3 813.2 1437.2
3m 2-CH3C6H4 C22H14O2Cl2 381.26 162-164 107.9 1136.4 1513.3
3n 3-CH3C6H4 C22H14O2Cl2 381.26 172-173 397.9 110.9 1199.2
3o 4-CH3C6H4 C22H14O2Cl2 381.26 204-206 11.2 865.2 11639
3p 3-ClC6H4 C21H11O2Cl3 401.67 172-174 11.7 1050.6 19840
3q 4-ClC6H4 C21H11O2Cl3 401.67 169-171 b b b
3r 4-NO2C6H4 C21H11NO4Cl2 412.22 198-200 b b b
3s 3-NO2C6H4 C21H11NO4Cl2 412.22 216-218 11.1 836.6 1481.7
3t 4-Cl C22H13O3Cl3 431.70 195-196 10.8 818.1 114.5
2-CH3O-C6H3
3u 2,4-Cl2C6H3 C22H10O2Cl4 436.12 190-192 87.7 107.5 1182.7
3v CH2C6H5 C22H14O2Cl2 381.25 152-154 109.8 12.6 1186.7
3w CH2CH2C6H5 C23H16O2Cl2 395.28 126-128 1734.8 936.3 1164.7
Doxorubicin 598.9 420.5 498.8
Mitomycin-C 1896.3 11720 2122.5
Methotrexate 9286.2 47180 9141.5

[0056]

TABLE 2
Taq inhibition data for novel 1,5-dichloro-anthracenes
Comp'd Taq Inhibition
No. R group 1 mM 0.1 mM 0.01 mM
3b CH2Br + + +
3c CH2Cl
3d CH2CH3 + + +
3g CHCl2 + + +
3h (CH2)2CH3 + + +
3i (CH2)3Br
3j (CH2)3Cl + +
3k (CH2)4CH3 + + +
3m 2-CH3C6H4
3n 3-CH3C6H4 + +
3p 3-ClC6H4 +
3q 4-ClC6H4 +
3r 3-NO2C6H4
3s 4-NO2C6H4
3u 2,4-Cl2C6H3 + +
3v CH2C6H5 + +
3w CH2CH2C6H5 + + +

[0057]

TABLE 3
Inhibitory effects of novel 1,5-dichloro-anthracene compounds of the
invention on iron-induced lipid peroxidation in rat brain homogenates
% Inhibitiona % Inhibitiona
Comp'd No. R group (10 μM) (1 μM)
3a CH3 34.4 25.5
3b CH2Br 100 100
3c CH2Cl 100 100
3d CH2CH3 100 100
3e CH(CH3)2 28.6 1.5
3f CH(CH3)Cl 60.6 9.8
3g CHCl2 100 61.7
3h (CH2)2CH3 17.7 11.8
3i (CH2)3Br 100 38.1
3j (CH2)3Cl 29.0 10.0
3k (CH2)4CH3 17.2 12.2
3L C6H5 14.0 4.6
3m 2-CH3C6H4 23.0 0
3n 3-CH3C6H4 22.4 10.9
3o 4-CH3C6H4 25.6 2.9
3p 3-ClC6H4 30.6 8.6
3q 4-ClC6H4 b b
3r 4-NO2C6H4 b b
3s 3-NO2C6H4 35.7 5.9
3t 4-Cl, 10.3 0
2-CH3O-C6H3
3u 2,4-Cl2C6H3 16.0 1.7
3v CH2C6H5 32.8 13.7
3w CH2CH2C6H5 36.6 29.3
(+)-a-Tocopherol 100 61.6

Referenced by
Citing PatentFiling datePublication dateApplicantTitle
US7822485 *Sep 25, 2006Oct 26, 2010Zoll Circulation, Inc.Method and apparatus for spinal cooling
Classifications
U.S. Classification560/139
International ClassificationA61K31/222, C07C67/00, A61K31/216, C07C205/57, C07C69/00
Cooperative ClassificationC07C2103/00, C07C2103/24, C07C205/57, C07C69/76, C07C69/017, C07C67/00, A61K31/222, C07C69/63, C07C69/78, A61K31/216, C07C69/92
European ClassificationC07C67/00, C07C69/78, C07C69/017, C07C69/92, C07C69/76, C07C69/63, A61K31/222, C07C205/57, A61K31/216
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