Substituted alkanohydroxamic acids and method of reducing TNF.alpha. levels Number:7,091,356 from the United States Patent and Trademark Office (PTO) owispatent Imido and Amido substituted alkanohydroxamic acids reduce the levels of TNFa and inhibit phosphodiesterase in a mammal. A typical embodiment is 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamide.<H alkanohydroxamic, Substituted, Substituted, alk, 7091356.html, Patent, pto, 7091356

Title: Substituted alkanohydroxamic acids and method of reducing TNF.alpha. levels

Abstract: Imido and Amido substituted alkanohydroxamic acids reduce the levels of TNFa and inhibit phosphodiesterase in a mammal. A typical embodiment is 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamide.

Patent Number: 7,091,356 Issued on 08/15/2006 to Muller, et al.

Inventors: Muller; George W. (Bridgewater, NJ), Man; Hon-Wah (Neshanic Station, NJ)
Appl. No.: 10/462,319

Filed: June 16, 2003

Related U.S. Patent Documents


Application Number	Filing Date	Patent Number	Issue Date
09780725	Feb., 2001	6656964
09126157	Jul., 1998	6214857
08903975	Jul., 1997

Current U.S. Class: 546/277.1 ; 548/302.4

Current International Class: C07D 401/00 (20060101)

Field of Search: 514/248,259,269,272,346,424,372 544/311 546/291 548/213,545

References Cited [Referenced By]

U.S. Patent Documents


4077998	March 1978	Fessler et al.
4173652	November 1979	Bruins et al.
4820828	April 1989	Demers et al.
6214857	April 2001	Muller et al.
6656964	December 2003	Muller et al.

Foreign Patent Documents


WO 97/05105	Feb., 1997	WO
WO 99/06041	Feb., 1999	WO

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Primary Examiner: Wang; Andrew
Assistant Examiner: Lundgren; Jeffrey S.
Attorney, Agent or Firm: Jones Day

Parent Case Text

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent application Ser. No. 09/780,725, filed Feb. 9, 2001, now U.S. Pat. No. 6,656,964, which is a divisional of U.S. patent application Ser. No. 09/126,157, filed Jul. 30, 1998, now U.S. Pat. No. 6,214,857, which is a continuation-in-part of U.S. patent application Ser. No. 08/903,975, filed Jul. 31, 1997, now abandoned.

Claims

The invention claimed is:

1. A substantially chirally pure (R)- or a substantially chirally pure (S)-, or mixture of (R)- and (S)-hydroxamic acid derivative selected from the group consisting of (a) compounds of the formula: ##STR00006## wherein each of R.sup.1 and R.sup.2, when taken independently of each other, is hydrogen, lower alkyl, or R.sup.1 and R.sup.2, when taken together, together with the depicted carbon atoms to which each is bound, is o-phenylene, o-naphthylene, or cyclohexene-1,2-diyl, unsubstituted or substituted with 1 to 4 substituents each selected independently from the group consisting of nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, cycloalkoxy of 3 to 6 carbon atoms, C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.3 C.sub.10-alkylidenemethyl, indanyloxy, and halo; R.sup.3 is phenyl substituted with from one to four substituents selected from the group consisting of nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, cycloalkoxy of 3 to 6 carbon atoms, and halo; or C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.3 C.sub.10-alkylidenemethyl; and R.sup.4 is hydrogen, alkyl of 1 to 10 carbon atoms, phenyl, or benzyl; R.sup.4' is hydrogen or alkyl of 1 to 6 carbon atoms; R.sup.5 is --CH.sub.2--, --CH.sub.2--CO--, --CO--, --SO.sub.2--, --S--, or --NHCO--; and n has a value of 0, 1, or 2; and (b) the acid addition salts of said compounds which contain a nitrogen atom capable of being protonated.

2. A hydroxamic acid derivative according to claim 1 wherein said compound has the formula: ##STR00007## in which each of R.sup.4 and R.sup.4 independently is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.5 is --CH.sub.2--, --CH.sub.2--CO--, or --CO--; each of R.sup.6 and R.sup.7, independently of the other, is nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, cylcoalkoxy of 3 to 6 carbon atoms, or halo; each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is hydrogen, nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, halo; and n has a value of 1.

3. A substantially chirally pure (R)- or a substantially chirally pure (S)-, or mixture of (R)- and (S)-hydroxamic acid derivative wherein said compound has the formula: ##STR00008## in which R.sup.4' is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.5 is C.dbd.O or CH.sub.2; n=1; each of R.sup.12 and R.sup.13, independently of each other is selected from alkoxy of 1 to 4 carbon atoms, cylcoalkoxy of 3 to 6 carbon atoms; C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.2 C.sub.10-alkylidenemethyl, or indanyloxy; and each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is hydrogen, nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms or halo.

4. The hydroxamic acid derivative according to claim 3 wherein R.sup.4' is Hydrogen.

5. A hydroxamic acid derivative according to claim 1 wherein said compound is selected from the group consisting of 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(1-oxoisoindolinyl)propionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-methoxy-3-(1-oxoisoindolinyl)propionamide, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-phthalimidopropionamide, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(3-nitrophthalimido)propionami- de, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(1-oxoisoindolinyl)propiona- mide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamide, N-hydroxy-3-(3,4-dimethoxyphenyl)-3-phthalimidopropionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(3-nitrophthalimido)propionamide- , N-hydroxy-3-(3,4-dimethoxyphenyl)-3-(1-oxoisoindolinyl)propionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(4-methylphthalimido)propionamid- e, 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamid- e, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(1,3-dioxo-2,3-dihydro-1H-benz- o[f]isoindol-2-yl)propionamide, N-hydroxy-3-{3-(2-propoxy)-4-methoxyphenyl}-3-phthal-imidopropionamide, 3-(3-ethoxy-4-methoxyphenyl)-3-(3,6-difluorophthalimido)-N-hydroxypropion- amide, 3-(4-aminophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropio- namide, 3-(3-aminophthalimido)-3-(3-methoxy-4-methoxyphenyl)-N-hydroxyprop- ionamide, 3-(3-aminophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypro- pionamide, 3-(3-aminophthalimido)-3-(3-cyclopentoxy-4-methoxyphenyl)-N-hyd- roxypropionamide, N-hydroxy-3-(3,4-dimethoxyphenyl)-3-(1-oxoisoindolinyl)propionamide, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(3-nitrophthalimido)propionami- de; 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-(1-oxoisoindolinyl)pr- opionamide, 3-(3-methylphthalimido)-3-(3-cyclopentoxy-4-methoxyphenyl)-N-hydroxypropi- onamide, 3-(4-methylphthalimido)-3-(3-cyclopentoxy-4-methoxyphenyl)-N-hydr- oxypropionamide, 3-(3-hydroxyphthalimido)-3-(3-cyclopentoxy-4-methoxyphenyl)-N-hydroxyprop- ionamide, 3-(4-hydroxyphthalimido)-3-(3-cyclopentoxy-4-methoxyphenyl)-N-hy- droxypropionamide, 3-(3-methylphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropionamid- e, 3-(4-methylphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropionam- ide, 3-(3-hydroxyphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropio- namide, 3-(4-hydroxyphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypro- pionamide, N-Hydroxy-N-methyl-3-(3-ethoxy-4-methoxyphenyl)-3-(1-oxoisoindo- linyl)propionamide, 3-(3-Cyclo-pentyloxy-4-methoxyphenyl)-N-hydroxy-3-(4-ethylphthalimido)pro- pionamide, 3-(3-Eth-oxy-4-methoxyphenyl)-N-hydroxy-3-(3-hydroxyphthalimido- )propionamide, 3-(3-Ethoxy-4-methoxyphenyl)-N-hydroxy-3-(4-hydroxyphthalimido)propionami- de, 3-(3-Ethoxy-4-methoxyphenyl)-N-hydroxy-3-(3-methylphthalimido)propiona- mide, 3-(3-Acetoamidophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypr- opionamide, 3-(4-Acetoamidophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropion- amide, 3-(3-Ethoxy-4-methoxyphenyl)-N-hydroxy-3-(1,3-dioxo-2,3-dihydro-1H-- enzo[e]isoindol-2'-yl)propionamide, 3-(4-tert-Butylphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropion- amide, 3-(3,4-dimethoxyphenyl)-N-hydroxy-3-(1,3-dioxo-2,3-dihydro-1H-benzo- [e]-isoindol-2'-yl)propionamide, 3-(3,4-Dimethoxyphthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropio- namide, 3-(3-Ethoxy-4-methoxyphenyl)-N-hydroxy-3-(3-dimethylaminophthalimi- do)-propionamide, 3-(6,8-Dioxo(2H-1,3-dioxolano[4,5-e]isoindolin-7-yl))-3-(3-ethoxy-4-metho- xyphenyl)-N-hydroxypropionamide, and 3-(3-Ethoxy-4-methoxyphenyl)-N-hydroxy-3-(3,4-dimethylphthalimido)propion- amide.

6. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 1 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regimen in combination with a carrier.

7. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 1 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

8. A method of reducing undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 1 in combination with a carrier.

9. A method of inhibiting undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 1 in combination with a suitable carrier.

10. The hydroxamic acid derivative of claim 2, wherein each of R.sup.8 and R.sup.11, independently of the others, is halo, N-acylamino or hydrogen and R.sup.5 is CO or CH.sub.2.

11. The hydroxamic acid derivative of claim 1, wherein R.sup.5 is CH.sub.2.

12. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 1 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

13. A method of treating cancer in a mammal which comprises administering an effective amount of a hydroxamic acid derivative according to claim 1.

14. The hydroxamic acid derivative of claim 2, wherein R.sup.5 is --CH2- or --CO--; and R.sup.6 and R.sup.7, independently of the other is alkoxy of 1 to 4 carbon atoms.

15. A method of reducing undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 2 in combination with a carrier.

16. A method of reducing undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 3 in combination with a carrier.

17. A method of reducing undesirable levels of TNF.alpha. in a mammal which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 2 in combination with a carrier.

18. A method of reducing undesirable levels of TNFa in a mammal which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 3 in combination with a carrier.

19. A method of inhibiting undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a suitable carrier which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 2.

20. A method of inhibiting undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a suitable carrier which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 3.

21. A method of treating cancer in a host which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 2.

22. A method of treating cancer in a host which comprises administering thereto an effective amount of a hydroxamic acid derivative according to claim 3.

23. The compound of claim 1 that is a substantially chirally pure (R)-hydroxamic acid derivative.

24. The compound of claim 1 that is a substantially chirally pure (S)-hydroxamic acid derivative.

25. The compound of claim 1 that is a mixture of (R)- and (S)-hydroxamic acid derivatives.

26. The compound of claim 3 that is a substantially chirally pure (R)-hydroxamic acid derivative.

27. The compound of claim 3 that is a substantially chirally pure (S)-hydroxamic acid derivative.

28. The compound of claim 3 that is a mixture of (R)- and (S)-hydroxamic acid derivatives.

29. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 3 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

30. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 3 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

31. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 3 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

32. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 23 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

33. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 23 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

34. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 23 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

35. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 24 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

36. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 24 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

37. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 24 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

38. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 25 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

39. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 25 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

40. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 25 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

41. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 26 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

42. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 26 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

43. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 26 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

44. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 27 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

45. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 27 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

46. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 27 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

47. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 28 sufficient upon administration to reduce undesirable levels of TNF.alpha. in a mammal in a single or multiple dose regiment in combination with a carrier.

48. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 28 sufficient upon administration to inhibit undesirable levels of matrix metalloproteinases in a mammal in a single or multiple dose regimen in combination with a carrier.

49. A pharmaceutical composition comprising an effective amount of a hydroxamic acid derivative according to claim 28 sufficient upon administration to inhibit PDE4 and MMP in a mammal in a single or multiple dose regimen in combination with a carrier.

Description

FIELD OF THE INVENTION

The present invention relates to imido and amido substituted alkanohydroxamic acids, the method of reducing levels of tumor necrosis factor a in a mammal through the administration thereof, and pharmaceutical compositions of such derivatives.

BACKGROUND OF THE INVENTION

Tumor necrosis factor .alpha., or TNF.alpha., is a cytokine which is released primarily by mononuclear phagocytes in response to a number immunostimulators. When administered to animals or humans, it causes inflammation, fever, cardiovascular effects, hemorrhage, coagulation, and acute phase responses similar to those seen during acute infections and shock states. Excessive or unregulated TNF.alpha. production thus has been implicated in a number of disease conditions. These include endotoxemia and/or toxic shock syndrome {Tracey et al., Nature 330, 662 664 (1987) and Hinshaw et al., Circ. Shock 30, 279 292 (1990)}; cachexia {Dezube et al., Lancet, 335 (8690), 662 (1990)} and Adult Respiratory Distress Syndrome where TNF.alpha. concentration in excess of 12,000 pg/mL have been detected in pulmonary aspirates from ARDS patients {Millar et al., Lancet 2(8665), 712 714 (1989)}. Systemic infusion of recombinant TNF.alpha. also resulted in changes typically seen in ARDS {Ferrai-Baliviera et al., Arch. Surg. 124(12), 1400 1405 (1989)}.

TNF.alpha. appears to be involved in bone resorption diseases, including arthritis. When activated, leukocytes will produce bone-resorption, an activity to which the data suggest TNF.alpha. contributes. {Bertolini et al., Nature 319, 516 518 (1986) and Johnson et al., Endocrinology 124(3), 1424 1427 (1989).} TNF.alpha. also has been shown to stimulate bone resorption and inhibit bone formation in vitro and in vivo through stimulation of osteoclast formation and activation combined with inhibition of osteoblast function. Although TNF.alpha. may be involved in many bone resorption diseases, including arthritis, the most compelling link with disease is the association between production of TNF.alpha. by tumor or host tissues and malignancy associated hypercalcemia {Calci. Tissue Int. (US) 46(Suppl.), S3 10 (1990)}. In Graft versus Host Reaction, increased serum TNF.alpha. levels have been associated with major complication following acute allogenic bone marrow transplants {Holler et al., Blood, 75(4), 1011 1016 (1990)}.

Cerebral malaria is a lethal hyperacute neurological syndrome associated with high blood levels of TNF.alpha. and the most severe complication occurring in malaria patients. Levels of serum TNF.alpha. correlated directly with the severity of disease and the prognosis in patients with acute malaria attacks {Grau et al., N. Engl. J. Med. 320(24), 1586 1591 (1989)}.

Macrophage-induced angiogenesis TNF.alpha. is known to be mediated by TNF.alpha.. Leibovich et al. {Nature, 329, 630 632 (1987)} showed TNF.alpha. induces in vivo capillary blood vessel formation in the rat cornea and the developing chick chorioallantoic membranes at very low doses and suggest TNF.alpha. is a candidate for inducing angiogenesis in inflammation, wound repair, and tumor growth. TNF.alpha. production also has been associated with cancerous conditions, particularly induced tumors {Ching et al., Brit. J. Cancer, (1955) 72, 339 343, and Koch, Progress in Medicinal Chemistry, 22, 166 242 (1985)}.

TNF.alpha. also plays a role in the area of chronic pulmonary inflammatory diseases. The deposition of silica particles leads to silicosis, a disease of progressive respiratory failure caused by a fibrotic reaction. Antibody to TNF.alpha. completely blocked the silica-induced lung fibrosis in mice {Pignet et al., Nature, 344, 245 247 (1990)}. High levels of TNF.alpha. production (in the serum and in isolated macrophages) have been demonstrated in animal models of silica and asbestos induced fibrosis {Bissonnette et al., Inflammation 13(3), 329 339 (1989)}. Alveolar macrophages from pulmonary sarcoidosis patients have also been found to spontaneously release massive quantities of TNF.alpha. as compared with macrophages from normal donors {Baughman et al., J. Lab. Clin. Med. 115(1), 36 42 (1990)}.

TNF.alpha. is also implicated in the inflammatory response which follows reperfusion, called reperfusion injury, and is a major cause of tissue damage after loss of blood flow {Vedder et al., PNAS 87, 2643 2646 (1990)}. TNF.alpha. also alters the properties of endothelial cells and has various pro-coagulant activities, such as producing an increase in tissue factor pro-coagulant activity and suppression of the anticoagulant protein C pathway as well as down-regulating the expression of thrombomodulin {Sherry et al., J. Cell Biol. 107, 1269 1277 (1988)}. TNF.alpha. has pro-inflammatory activities which together with its early production (during the initial stage of an inflammatory event) make it a likely mediator of tissue injury in several important disorders including but not limited to, myocardial infarction, stroke and circulatory shock. Of specific importance may be TNF.alpha.-induced expression of adhesion molecules, such as intercellular adhesion molecule (ICAM) or endothelial leukocyte adhesion molecule (ELAM) on endothelial cells {Munro et al., Am. J. Path. 135(1), 121 132 (1989)}.

TNF.alpha. blockage with monoclonal anti-TNF.alpha. antibodies has been shown to be beneficial in rheumatoid arthritis {Elliot et al., Int. J. Pharmac. 1995 17(2), 141 145)}. High levels of TNF.alpha. are associated with Crohn's disease {von Dullemen et al., Gastroenterology, 1995 109(1), 129 135} and clinical benefit has been achieved with TNF.alpha. antibody treatment.

Moreover, it now is known that TNF.alpha. is a potent activator of retrovirus replication including activation of HIV-1. {Duh et al., Proc. Nat. Acad. Sci. 86, 5974 5978 (1989); Poll et al., Proc. Nat. Acad. Sci. 87, 782 785 (1990); Monto et al., Blood 79, 2670 (1990); Clouse et al., J. Immunol. 142, 431 438 (1989); Poll et al., AIDS Res. Hum. Retrovirus, 191 197 (1992)}. AIDS results from the infection of T lymphocytes with Human Immunodeficiency Virus (HIV). At least three types or strains of HIV have been identified, i.e., HIV-1, HIV-2 and HIV-3. As a consequence of HIV infection, T-cell mediated immunity is impaired and infected individuals manifest severe opportunistic infections and/or unusual neoplasms. HIV entry into the T lymphocyte requires T lymphocyte activation. Other viruses, such as HIV-1, HIV-2 infect T lymphocytes after T cell activation and such virus protein expression and/or replication is mediated or maintained by such T cell activation. Once an activated T lymphocyte is infected with HIV, the T lymphocyte must continue to be maintained in an activated state to permit HIV gene expression and/or HIV replication. Cytokines, specifically TNF.alpha., are implicated in activated T-cell mediated HIV protein expression and/or virus replication by playing a role in maintaining T lymphocyte activation. Therefore, interference with cytokine activity such as by prevention or inhibition of cytokine production, notably TNF.alpha., in an HIV-infected individual assists in limiting the maintenance of T lymphocyte caused by HIV infection.

Monocytes, macrophages, and related cells, such as kupffer and glial cells, also have been implicated in maintenance of the HIV infection. These cells, like T cells, are targets for viral replication and the level of viral replication is dependent upon the activation state of the cells. {Rosenberg et al., The Immunopathogenesis of HIV Infection, Advances in Immunology, 57 (1989)}. Cytokines, such as TNF.alpha., have been shown to activate HIV replication in monocytes and/or macrophages {Poli et al., Proc. Natl. Acad. Sci., 87, 782 784 (1990)}, therefore, prevention or inhibition of cytokine production or activity aids in limiting HIV progression for T cells. Additional studies have identified TNF.alpha. as a common factor in the activation of HIV in vitro and has provided a clear mechanism of action via a nuclear regulatory protein found in the cytoplasm of cells (Osborn, et al., PNAS 86 2336 2340). This evidence suggests that a reduction of TNF.alpha. synthesis may have an antiviral effect in HIV infections, by reducing the transcription and thus virus production.

AIDS viral replication of latent HIV in T cell and macrophage lines can be induced by TNF.alpha. {Folks et al., PNAS 86, 2365 2368 (1989)}. A molecular mechanism for the virus inducing activity is suggested by TNF.alpha.'s ability to activate a gene regulatory protein (NF.kappa.B) found in the cytoplasm of cells, which promotes HIV replication through binding to a viral regulatory gene sequence (LTR) {Osborn et al., PNAS 86, 2336 2340 (1989)}. TNF.alpha. in AIDS associated cachexia is suggested by elevated serum TNF.alpha. and high levels of spontaneous TNF.alpha. production in peripheral blood monocytes from patients {Wright et al., J. Immunol. 141(1), 99 104 (1988)}. TNF.alpha. has been implicated in various roles with other viral infections, such as the cytomegalia virus (CMV), influenza virus, adenovirus, and the herpes family of viruses for similar reasons as those noted.

The nuclear factor .kappa.B (NF.kappa.B) is a pleiotropic transcriptional activator (Lenardo, et al., Cell 1989, 58, 227 29). NF.kappa.B has been implicated as a transcriptional activator in a variety of disease and inflammatory states and is thought to regulate cytokine levels including but not limited to TNF.alpha. and also to be an activator of HIV transcription (Dbaibo, et al., J. Biol. Chem. 1993, 17762 66; Duh et al., Proc. Natl. Acad. Sci. 1989, 86, 5974 78; Bachelerie et al., Nature 1991, 350, 709 12; Boswas et al., J. Acquired Immune Deficiency Syndrome 1993, 6, 778 786; Suzuki et al., Biochem. And Biophys. Res. Comm. 1993, 193, 277 83; Suzuki et al., Biochem. And Biophys. Res Comm. 1992, 189, 1709 15; Suzuki et al., Biochem. Mol. Bio. Int. 1993, 31(4), 693 700; Shakhov et al., Proc. Natl. Acad. Sci. USA 1990, 171, 35 47; and Staal et al., Proc. Natl. Acad. Sci. USA 1990, 87, 9943 47). Thus, inhibition of NF.kappa.B binding can regulate transcription of cytokine gene(s) and through this modulation and other mechanisms be useful in the inhibition of a multitude of disease states. The compounds described herein can inhibit the action of NF.kappa.B in the nucleus and thus are useful in the treatment of a variety of diseases including but not limited to rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, other arthritic conditions, septic shock, septis, endotoxic shock, graft versus host disease, wasting, Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythrematosis, ENL in leprosy, HIV, AIDS, and opportunistic infections in AIDS. TNF.alpha. and NF.kappa.B levels are influenced by a reciprocal feedback loop. As noted above, the compounds of the present invention affect the levels of both TNF.alpha. and NF.kappa.B.

Many cellular functions are mediated by levels of adenosine 3',5'-cyclic monophosphate (cAMP). Such cellular functions can contribute to inflammatory conditions and diseases including asthma, inflammation, and other conditions (Lowe and Cheng, Drugs of the Future, 17(9), 799 807, 1992). It has been shown that the elevation of cAMP in inflammatory leukocytes inhibits their activation and the subsequent release of inflammatory mediators, including TNF.alpha. and NF.kappa.B. Increased levels of cAMP also leads to the relaxation of airway smooth muscle. Phosphodiesterases control the level of cAMP through hydrolysis and inhibitors of phosphodiesterases have been shown to increase cAMP levels.

Decreasing TNF.alpha. levels and/or increasing cAMP levels thus constitutes a valuable therapeutic strategy for the treatment of many inflammatory, infectious, immunological or malignant diseases. These include but are not restricted to septic shock, sepsis, endotoxic shock, hemodynamic shock and sepsis syndrome, post ischemic reperfusion injury, malaria, mycobacterial infection, meningitis, psoriasis, congestive heart failure, fibrotic disease, cachexia, graft rejection, cancer, autoimmune disease, opportunistic infections in AIDS, rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, other arthritic conditions, Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythrematosis, ENL in leprosy, radiation damage, and hyperoxic alveolar injury. Prior efforts directed to the suppression of the effects of TNF.alpha. have ranged from the utilization of steroids such as dexamethasone and prednisolone to the use of both polyclonal and monoclonal antibodies {Beutler et al., Science 234, 470 474 (1985); WO 92/11383}.

Matrix metalloproteinase (MMP) inhibition has been associated with inhibition of TNF. Mohler et al., Nature, 370, 218 220 (1994). MMPs, or matrixins, are a family of secreted and membrane-bound zinc endopeptidases that play a key role in both physiological and pathological tissue degradation. See Yu et al., Drugs & Aging, 1997, (3):229 244; Wojtowicz-Praga et al., Int. New Drugs, 16:61 75 (1997). These enzymes are capable of degrading the components of the extracellular matrix, including fibrillar and non-fibrillar collagens, fibronectin, laminin, and membrane glycoproteins. Ordinarily, there is a delicate balance between cell division, matrix synthesis, and matrix degradation (under the control of cytokines), growth factors, and cell matrix interactions. Under pathological conditions, however, this balance can be disrupted. Conditions and diseases associated with undesired MMP levels include, but are not limited to, tumor metastasis, invasion, and growth, rheumatoid arthritis, osteoarthritis, osteopenias such as osteoporosis, periodontitis, gingivitis, and corneal epidermal or gastric ulceration.

Increased MMP activity has been detected in a wide range of cancers, Denis et al., Invest. New Drugs, 15: 175 185 (1987), and as with TNF, MMPs are believed to be involved in the invasive processes of angiogenesis and tumor metastasis.

DETAILED DESCRIPTION

The present invention is based on the discovery that certain classes of non-polypeptide compounds more fully described herein decrease the levels of TNF.alpha., increase cAMP levels, and inhibit phosphodiesterase.

In particular, the invention pertains to

(a) compounds of the formula:

##STR00001## R.sup.4' is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.5 is C.dbd.O or CH.sub.2; each of R.sup.12 and R.sup.13, independently of the other alkoxy of 1 to 4 carbon atoms, cylcoalkoxy of 3 to 6 carbon atoms; C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.2 C.sub.10-alkylidenemethyl, C.sub.6 C.sub.10-bicycloalkoxy, or indanyloxy; each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is (i) hydrogen, nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, and halo, or (ii) hydrogen if one of the R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is acylamino comprising a lower alkyl, or (iii) hydrogen if the R.sup.10 and R.sup.11 taken together are benzo, methylenedioxy, or dioxo, or (iv) hydrogen if R.sup.9 and R.sup.10 taken together are benzo; and

(b) the acid addition salts of said compounds which contain a nitrogen atom capable of being protonated.

Subgroups of Formula I can include the following: The hydroxyamic acid derivative in Formula I, wherein R.sup.4' is hydrogen; R.sup.5 is C.dbd.O; R.sup.8 is hydrogen; and one of R.sup.9 and R.sup.11 is hydrogen and the other of R.sup.9 and R.sup.11, taken together with R.sup.10, is benzo. The hydroxyamic acid derivative in Formula I, wherein R.sup.4' is hydrogen; R.sup.5 is C.dbd.O; R.sup.8 and R.sup.9 are hydrogen; and R.sup.10 and R.sup.11, taken together, are methylenedioxo or dioxo. The hydroxyamic acid derivative in Formula I, wherein each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is hydrogen, alkyl of 1 to 10 carbon atoms, or alkoxy of 1 to 10 carbon atoms. The hydroxyamic acid derivative in Formula I, wherein R.sup.8, R.sup.9, R.sup.10, and R.sup.11 are (a) at least one alkyl of 1 to 10 carbon atoms with the remainder of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 being hydrogen, or (b) at least one alkoxy of 1 to 10 carbon atoms with the remainder of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 being hydrogen. The alkanohydroxyamic acid derivative in Formula I, which is a substantially chirally pure (3S)-isomer, a substantially chirally pure (3R)-isomer, or mixtures thereof. The hydroxamic acid derivative in Formula I, wherein R.sup.4' is hydrogen. The hydroxamic acid derivative in Formula I, wherein each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is hydrogen, halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms. A hydroxamic acid derivative in Formula I, wherein one of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is -amino, hydroxy, or alkyl and the remaining of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, are hydrogen.

Unless otherwise defined, the term alkyl denotes a univalent saturated branched or straight hydrocarbon chain containing from 1 to 8 carbon atoms. Representative of such alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl. Alkoxy refers to an alkyl group bound to the remainder of the molecule through an ethereal oxygen atom. Representative of such alkoxy groups are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

This invention also pertains to

(a) compounds of Formula I

wherein

each of R.sup.1 and R.sup.2, when taken independently of each other, is hydrogen, lower alkyl, or R.sup.1 and R.sup.2, when taken together, together with the depicted carbon atoms to which each is bound, is o-phenylene, o-naphthylene, or cyclohexene-1,2-diyl, unsubstituted or substituted with 1 to 4 substituents each selected independently from the group consisting of nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, and halo; R.sup.3 is phenyl substituted with from one to four substituents selected from the group consisting of nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, alkylthio of 1 to 10 carbon atoms, benzyloxy, cycloalkoxy of 3 to 6 carbon atoms, C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.3 C.sub.10-alkylidenemethyl, indanyloxy, and halo; R.sup.4 is hydrogen, alkyl of 1 to 6 carbon atoms, phenyl, or benzyl; R.sup.4' is hydrogen or alkyl of 1 to 6 carbon atoms; R.sup.5 is is --CH.sub.2--, --CH.sub.2--CO--, --CO--, --SO.sub.2--, --S--, or --NHCO--; and n has a value of 0, 1, or 2; and (b) the acid addition salts of said compounds which contain a nitrogen atom capable of being protonated.

The compounds of Formula I are used, under the supervision of qualified professionals, to inhibit the undesirable effects of TNF.alpha. and inhibit phosphodiesterase. The compounds can be administered orally, rectally, or parenterally, alone or in combination with other therapeutic agents including antibiotics, steroids, etc., to a mammal in need of treatment.

The compounds of the present invention also can be used topically in the treatment or prophylaxis of topical disease states mediated or exacerbated by excessive TNF.alpha. production, respectively, such as viral infections, such as those caused by the herpes viruses, or viral conjunctivitis, psoriasis, atopic dermatitis, etc.

The compounds also can be used in the veterinary treatment of mammals other than humans in need of prevention or inhibition of TNF.alpha. production. TNF.alpha. mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted above, but in particular viral infections. Examples include feline immunodeficiency virus, equine infectious anaemia virus, caprine arthritis virus, visna virus, and maedi virus, as well as other lentiviruses.

Angiogenesis, the process of new blood vessel development and formation, plays an important role in numerous physiological events, both normal and pathological. The compounds also can be used to inhibit unwanted angiogenesis.

The invention also relates to MMP-inhibiting compounds, compositions thereof, and their use in the treatment of diseases and disorders associated with undesired production or activity of MMPs. These compounds are capable of inhibiting connective tissue breakdown, and are useful in the treatment or prevention of conditions involving tissue breakdown. These include, but are not limited to, tumor metastasis, invasion, and growth, rheumatoid arthritis, osteoarthritis, osteopenias such as osteoporosis, periodontitis, gingivitis, and corneal epidermal or gastric ulceration.

The invention thus further comprises MMP-inhibiting compounds of Formula I, and methods of treatment comprising administering an effective amount of a compound according to Formula I.

The compounds of Formula I are readily prepared by reacting a carboxylic acid of the formula:

##STR00002## in which each of R.sup.1, R.sup.2, R.sup.3, R.sup.5, and n are as defined above, with hydroxylamine hydrochloride or alkoxyamine hydrochloride in the presence of a coupling agent. The reaction generally is conducted in an inert solvent such as tetrahydrofuran under an inert atmosphere such as nitrogen. Ambient temperatures can be employed. When the reaction is substantially complete, the products can be readily isolated simply through the addition of water.

The compounds of Formula II which are here utilized as intermediates are described in U.S. Pat. No. 5,605,914, the disclosure of which is incorporated herein by reference. Briefly, such intermediates can be prepared through the reaction of an amino acid of the formula:

##STR00003## in which R.sup.14 is hydroxy or a protecting group, with an acid anhydride, an N-carbethoxyimide, a dialdehyde, or an o-bromo aromatic acid. Protecting groups utilized herein denote groups which generally are not found in the final therapeutic compounds but which are intentionally introduced at some stage of the synthesis in order to protect groups which otherwise might be altered in the course of chemical manipulations. Such protecting groups are removed at a later stage of the synthesis and compounds bearing such protecting groups thus are of importance primarily as chemical intermediates (although some derivatives also exhibit biological activity). Accordingly the precise structure of the protecting group is not critical. Numerous reactions for the formation and removal of such protecting groups are described in a number of standard works including, for example, "Protective Groups in Organic Chemistry", Plenum Press, London and New York, 1973; Greene, Th. W. "Protective Groups in Organic Synthesis", Wiley, N.Y., 1981; "The Peptides", Vol. I, Schroder and Lubke, Academic Press, London and New York, 1965; "Methoden der organischen Chemie", Houben-Weyl, 4th Edition, Vol. 15/I, Georg Thieme Verlag, Stuttgart 1974, the disclosures of which are incorporated herein by reference.

In any of the foregoing reactions, a nitro compound can be employed with the nitro group being converted to an amino group by catalytic hydrogenation. Alternatively, a protected amino group can be cleaved to yield the corresponding amino compound. An amino group can be protected as an amide utilizing an acyl group which is selectively removable under mild conditions, especially benzyloxycarbonyl, formyl, or a lower alkanoyl group which is branched in 1- or .alpha. position to the carbonyl group, particularly tertiary alkanoyl such as pivaloyl, a lower alkanoyl group which is substituted in the position .alpha. to the carbonyl group, as for example trifluoroacetyl.

The compounds of Formula I possess at least one center of chirality (designated by "*") and can exist as optical isomers. Both the racemates of these isomers and the individual isomers themselves, as well as diastereomers when there are two chiral centers, are within the scope of the pres??ent invention. The racemates can be used as such or can be separated into their individual isomers mechanically as by chromatography using a chiral absorbant. Alternatively, the individual isomers can be prepared in chiral form or separated chemically from a mixture by forming salts with a chiral acid or base, such as the individual enantiomers of 10-camphorsulfonic acid, camphoric acid, .alpha.-bromocamphoric acid, methoxyacetic acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid, and the like, and then freeing one or both of the resolved bases, optionally repeating the process, so as obtain either or both substantially free of the other; i.e., in a form having an optical purity of >95%.

The present invention also pertains to the physiologically acceptable non-toxic acid addition salts of the compound of Formula I. Such salts include those derived from organic and inorganic acids such as, without limitation, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulphonic acid, acetic acid, tartaric acid, lactic acid, succinic acid, citric acid, malic acid, maleic acid, sorbic acid, aconitic acid, salicylic acid, phthalic acid, embonic acid, enanthic acid, and the like.

The present invention also pertains to the physiologically acceptable non-toxic salts of the compound of Formula I with bases such as the sodium salt, the potassium salt, the aluminum salt, and the like.

A first preferred subgroup encompasses compounds of the formula:

##STR00004## in which R.sup.4 is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.4' is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.5 is C.dbd.O or CH.sub.2; each of R.sup.6 and R.sup.7, independently of the other, is nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, cylcoalkoxy of 3 to 6 carbon atoms, bicycloalkyl of 6 to 10 carbon atoms, indanyloxy, C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.3 C.sub.10-alkylidenemethyl, or halo;

each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is hydrogen, nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, and halo; and

n has a value of 1.

Among the compounds of Formula IV, those in which each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is hydrogen, halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms, and those in which one of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is amino, hydroxy, or methyl, and the remaining of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, are hydrogen are particularly preferred.

A further preferred subgroup encompasses compounds of the formula:

##STR00005## in which R.sup.4' is hydrogen or alkyl of 1 to 4 carbon atoms; R.sup.5 is C.dbd.O or CH.sub.2; each of R.sup.12 and R.sup.13, independently of the other alkoxy of 1 to 4 carbon atoms, cylcoalkoxy of 3 to 6 carbon atoms; C.sub.4 C.sub.6-cycloalkylidenemethyl, C.sub.3 C.sub.10-alkylidenemethyl, or indanyloxy; and each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, independently of the others, is hydrogen, nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkylamino, dialkylamino, acylamino, alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, and halo.

Among the compounds of Formula V, those in which each of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is hydrogen, halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms, and those in which one of R.sup.8, R.sup.9, R.sup.10, and R.sup.11 is amino or hydroxy and the remaining of R.sup.8, R.sup.9, R.sup.10, and R.sup.11, are hydrogen are particularly preferred.

Particularly preferred compounds include 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(1-oxoisoindolinyl)propionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-methoxy-3-(1-oxoisoindolinyl)propionamide, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-phthalimidopropionamide, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(3-nitrophthalimido)propionami- de, N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(1-oxoisoindolinyl)-propion- amide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamide, N-hydroxy-3-(3,4-dimethoxyphenyl)-3-phthalimidopropionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(3-nitrophthalimido)propionamide- , N-hydroxy-3-(3,4-dimethoxyphenyl)-3-(1-oxoisoindolinyl)propionamide, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(4-methylphthalimido)propionamid- e, 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-phthalimidopropionamid- e, 3-(3-ethoxy-4-methoxyphenyl)-N-hydroxy-3-(1,3-dioxo-2,3-dihydro-1H-benz- o[f]isoindol-2-yl)propionamide, N-hydroxy-3-{3-(2-propoxy)-4-methoxyphenyl}-3-phthalimidopropionamide, 3-(3-ethoxy-4-methoxyphenyl)-3-(3,6-difluorophthalimido)-N-hydroxypropion- amide, 3-(4-aminophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropio- namide, 3-(3-aminophthalimido)-3-(3-ethoxy-4-methoxyphenyl)-N-hydroxypropi- onamide, N-hydroxy-3-(3,4-dimethoxyphenyl)-3-(1-oxoisoindolinyl)propionami- de, 3-(3-cyclopentyloxy-4-methoxyphenyl)-N-hydroxy-3-(1-oxoisoindolinyl)pr- opionamide, and N-benzyloxy-3-(3-ethoxy-4-methoxyphenyl)-3-(3-nitrophthalimido)propionami- de.

Oral dosage forms include tablets, capsules, dragees, and similar shaped, compressed pharmaceutical forms containing from 1 to 100 mg of drug per unit dosage. Isotonic saline solutions containing from 20 to 100 mg/mL can be used for parenteral administration which includes intramuscular, intrathecal, intravenous and intra-arterial routes of administration. Rectal administration can be effected through the use of suppositories formulated from conventional carriers such as cocoa butter.

Pharmaceutical compositions thus comprise one or more compounds of Formulas I associated with at least one pharmaceutically acceptable carrier, diluent or excipient. In preparing such compositions, the active ingredients are usually mixed with or diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule or sachet. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material which acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders. Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidinone, cellulose, water, syrup, and methyl cellulose, the formulations can additionally include lubricating agents such as talc, magnesium stearate and mineral oil, wetting agents, emulsifying and suspending agents, preserving agents such as methyl- and propylhydroxybenzoates, sweetening agents or flavoring agents.

The compositions preferably are formulated in unit dosage form, meaning physically discrete units suitable as a unitary dosage, or a predetermined fraction of a unitary dose to be administered in a single or multiple dosage regimen to human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient. The compositions can be formulated so as to provide an immediate, sustained or delayed release of active ingredient after administration to the patient by employing procedures well known in the art.

Enzyme-linked immunosorbent assays (ELISA) for TNF.alpha. can be performed in a conventional manner. PBMC is isolated from normal donors by Ficoll-Hypaque density centrifugation. Cells are cultured in RPMI supplemented with 10% AB+ serum, 2mM L-glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin. Drugs are dissolved in dimethylsulfoxide (Sigma Chemical) and further dilutions are done in supplemented RPMI. The final dimethylsulfoxide concentration in the presence or absence of drug in the PBMC suspensions is 0.25 wt %. Drugs are assayed at half-log dilutions starting at 50 mg/mL. Drugs are added to PBMC (10.sup.6 cells/mL) in 96 wells plates one hour before the addition of LPS. PBMC (10.sup.6 cells/mL) in the presence or absence of drug are stimulated by treatment with 1 mg/mL of LPS from Salmonella minnesota R595 (List Biological Labs, Campbell, Calif.). Cells are then incubated at 37.degree. C. for 18 20 hours. Supernatants are harvested and assayed immediately for TNF.alpha. levels or kept frozen at -70.degree. C (for not more than 4 days) until assayed. The concentration of TNF.alpha. in the supernatant is determined by human TNF.alpha. ELISA kits (ENDOGEN, Boston, Mass.) according to the manufacturer's directions.

Phosphodiesterase can be determined in conventional models. For example, using the method of Hill and Mitchell, U937 cells of the human promonocytic cell line are grown to 1.times.10.sup.6 cells/mL and collected by centrifugation. A cell pellet of 1.times.10.sup.9 cells is washed in phosphate buffered saline and then frozen at -70.degree. C. for later purification or immediately lysed in cold homogenization buffer (20 mM Tris-HCl, pH 7.1, 3 mM 2-mercaptoethanol, 1 mM magnesium chloride, 0.1 mM ethylene glycol-bis-(.beta.-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 1 .mu.M phenylmethylsulfonyl fluoride (PMSF), and 1 .mu.g/mL leupeptin). Cells are homogenized with 20 strokes in a Dounce homogenizer and supernatant containing the cytosolic fraction are obtained by centrifugation. The supernatant then is loaded onto a Sephacryl S-200 column equilibrated in homogenization buffer. Phosphodiesterase is eluted in homogenization buffer at a rate of approximately 0.5 mL/min and fractions are assayed for phosphodiesterase activity -/+ rolipram. Fractions containing phosphodiesterase activity (rolipram sensitive) are pooled and aliquoted for later use.

The phosphodiesterase assay is carried out based on procedure described by Hill and Mitchell. The assay is carried out in a total volume of 100 .mu.l containing various concentration of Celgene compounds, 50 mM Tris-HCl, pH 7.5,5 mM magnesium chloride and 1 .mu.M cAMP of which 1% was .sup.3H cAMP. Reactions are incubated at 30.degree. C. for 30 minutes and terminated by boiling for 2 minutes. The amount of phosphodiesterase IV containing extract used for these experiments is predetermined such that reactions are within the linear range and consumed less than 15% of the total substrate. Following termination of reaction, samples are chilled at 4.degree. C. and then treated with 10 .mu.l 10 mg/mL snake venom for 15 min at 30.degree. C. Unused substrate then is removed by adding 200 .mu.l of a quaternary ammonium ion exchange resin (AG1-X8, BioRad) for 15 minutes. Samples then are spun at 3000 rpm, 5 min and 50 .mu.l of the aqueous phase are taken for counting. Each data point is carried out in duplicate and activity is expressed as percentage of control. The IC.sub.50 of the compound then is determined from dose response curves of a minimum of three independent experiments.

The following examples will serve to further typify the nature of this invention but should no