`Muller et al.
`
`USOO628.1230B1
`(10) Patent No.:
`US 6,281,230 B1
`(45) Date of Patent:
`Aug. 28, 2001
`
`(54) ISOINDOLINES, METHOD OF USE, AND
`PHARMACEUTICAL COMPOSITIONS
`(75) Inventors: George W. Muller, Bridgewater; David
`I. Stirling, Branchburg; Roger
`Shen-Chu Chen, Edison, all of NJ
`(US)
`(73) Assignee: Celgene Corporation, Warren, NJ (US)
`
`(*) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`(21) Appl. No.: 09/543,809
`(22) Filed:
`Apr. 6, 2000
`
`O
`O
`Related U.S. Application Data
`(62) Division of application No. 09/230,389, filed as application
`No. PCT/US97/13375 on Jul. 24, 1997, now abandoned,
`which is a continuation of application No. 08/690,258, filed
`on Aug. 22, 1996, now Pat. No. 5,798.368, which is a
`initie? Epigg Ng 08/701,499, filed on Jul. 24,
`(60) Provisional application No. 60/048.278, filed on May 30,
`1997.
`
`(51) Int. Cl." ....................... A61K 31/445; CO7D 401/04
`
`(52) U.S. Cl. ............................................. 514/323:546/201
`(58) Field of Search .............................. 514323.546,201
`(56)
`References Cited
`
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`3/1996 Heinemann et al..
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`1/1997 D'Amato.
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`2/1997 Muller.
`5629.327
`5/1997 DAmato.
`5,635,517
`6/1997 Muller et al. ... s1432
`5,658,940
`8/1997 Muller et al. .
`5,698,579
`12/1997 Muller.
`5,703,098
`12/1997 Muller et al. .
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`al
`601050
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`6,020,358
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`6,046,221
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`6,071,948
`6/2000 DAmato.
`
`FOREIGN PATENT DOCUMENTS
`WO95/01348
`1/1995 (WO).
`
`OTHER PUBLICATIONS
`Corral, L. et al., “Differential Cytokine Modulation and T
`Cell Activation by Two Distinct Classes of Thalidomide
`Analogues That Are Potent Inhibitors of TNF- 1’, The
`Journal of Immunology, pp. 380-386, 1999.
`Corral, L. et al., “Immunomodulation by thalidomide and
`thalidomide analogues', Annals of the Rheumatic Diseases,
`vol. 58, Suppl. 1, pp. 1107–1113, 1999.
`Corral, L. et al., “Selection of Novel Analogs of Thalido
`mide with Enhanced Tumor Necrosis Factor Inhibitory
`Activity”, Molecular Medicine, vol. 2, No. 4, 1996.
`Craig, J.C., “Abcolute Configuration of the Enantiomers of
`7-Chloro-4-4-diethylamino)-1-methylbutylamino
`quinoline (chloroquine), J. Org. Chem., Vol. 53, pp.
`1167–1170, 1988.
`Feiser, Louis F., Experiments in Organic Chemistry, 3rd
`edition, p. 75, 1955.
`He, Weixuan et al., “Synthesis of Thalidomide Analogs and
`s Biological Potential for Treatment of Graft Versus
`ost Disease (GVHD)”, 206th American Chemical Society
`National Meeting, Med. Chem. Abst. 216, 1993.
`Koch, H., The Arene Oxide Hypothesis of Thalidomide
`Action. Considerations on the Molecular Mechanism of
`Action of the “Classical Teratogen, Sci. Pharm., vol. 49,
`pp. 67-99 (1981).
`Koch, H., 4 Thalidomide and Congeners as Anti-Inflamma
`tory Agents, ProgreSS in Medicinal Chemistry, Vol. 22, pp.
`166-242 (1985).
`Miyachi, H. et al., Novel Biological Response Modifiers:
`Phthalimides with Tumor Necrosis Factor- Production
`Regulating Activity, J. Med. Chem, pp. 2858-2865 (1997).
`Miyachi, H. et al., Tumor Necrosis Factor-Alpha Production
`Enhancing Activity of Substituted 3'-Methylthalidomide:
`Influence of Substituents at the Phthaloyl Moiety on the
`Activity of Stereoselectivity, Chem. Pharm. Bull., 46(7), pp.
`1165–1168 (1998).
`Muller, George et al., Amino-Substituted Thalidomide Ana
`logs: Potent Inhibitors of TNF- Production, Bioorganic &
`Medicinal Chem. Letters 9, pp. 1625–1630 (1999).
`Niwayama, Satomi et al., Potent Inhibition of Tumor Necro
`sis Factor- Production by Tetrafluorothalidomide and Tetr
`lfluorophtalimides, J. Med. Chem., pp. 3044-3045 (1996).
`Smith. R. L. et al.. Studies on the Relationship Between the
`s
`Ip
`Chemical Structure and Embryotoxic Activity of Thalido
`mide and Related Compounds, Symp. Embryopathic Act.
`Drugs, pp. 194-209 (1965).
`(List continued on next page.)
`Primary Examiner-Charanjit S. Aulakh
`(74) Attorney, Agent,
`or Firm-Mathews, Collins
`Sherpherd & Gould
`(57)
`ABSTRACT
`Substituted 1-oxo-2-(2,6-dioxopiperidin-3-yl)isoindolines
`are useful in treating inflammation, inflammatory disease,
`autoimmune disease, and oncogenic or cancerous conditions
`in a mammal. Typical embodiments are 1-oxo-2-(2,6-
`dioxopiperidin-3-yl)-4-aminoisoindoline and 1-oxo-2-(2,6-
`dioxo-3-methylpiperidin-3-yl)-4-aminoisoindoline.
`
`26 Claims, No Drawings
`
`ALVOGEN, Exh. 1028, p. 0001
`
`
`
`US 6,281.230 B1
`Page 2
`
`OTHER PUBLICATIONS
`Jonsson, N., Chenical Stucture and Teratogenic Properties,
`Acta. Pharm. Suicica, vol. 9, pp. 521-542 (1972).
`Muller, George et al., Stuctural Modifications of Thalido
`mide Produce Analogs with Enhanced Tumor Necrosis
`Factor Inhibitory Activity, Journal of Medicinal Chemistry,
`vol.39, No. 17, pp. 3238-3240(1996).
`Muller, George et al., Thalidomide Analogs and PDE4
`Inhibition, Bioorganic & Medicinal Chemistry Letters 8, pp.
`2669-2674 (1998).
`Niwayama, Satomi et al., Enhanced Potency of Perfluori
`nated Thalidomide Derivatives for Inhibition of LPS-In
`duced Tumor Necrosis Factor- Production is ASSociated
`with a Change of Mechanism of Action, Bioorganic &
`Medicinal Chemistry Letters 7, pp. 1071-1076 (1998).
`
`Shannon, Edward J. et al., Immunomodulatory ASSays to
`Study Structure-Activity Relationships of Thalidomide,
`Immunopharmacology 35, pp. 203-212 (1997).
`Takeuchi, Yoshio et al., (R)- and (S)-3-Fluorothalidomides:
`Isosteric Analogues of Thalidomide, American Chemical
`Society, vol. 1, No. 10, pp. 1571–1573 (1999).
`Udagawa, Taturo et al., Thalidomide and Analogs, Antian
`glogenia Agents in Cancer Therapy, pp. 263-274.
`Zwingenberger, K. et al., Immunomodulation by Thalido
`mide: Systematic Review of the Literature and of Unpub
`lished Observations, Journal of Inflammation, pp. 177-211
`(1996).
`
`* cited by examiner
`
`ALVOGEN, Exh. 1028, p. 0002
`
`
`
`1
`ISOINDOLINES, METHOD OF USE, AND
`PHARMACEUTICAL COMPOSITIONS
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`This is a divisional of Ser. No. 09/230,389, now
`abandoned, which is based on PCT/US97/13375 filed Jul.
`24, 1997, which is a continuation of Ser. No. 08/690,258
`filed Jul. 24, 1996, now U.S. Pat. No. 5,635,517, which is a
`continuation of Ser. No. 08/701,494 filed Aug. 22, 1996 now
`U.S. Pat. No. 5,798.368, and provisional application Ser.
`No. 60/048.278 filed May 30, 1997.
`
`DETAILED DESCRIPTION
`The present invention relates to substituted 2-(2,6-
`dioxopiperidin-3-yl)phthalimides and substituted 2-(2,6-
`dioxopiperidin-3-yl)-1-oxoisoindolines, the method of
`reducing levels of tumor necrosis factor C. in a mammal
`through the administration thereof, and pharmaceutical com
`positions of Such derivatives.
`
`15
`
`BACKGROUND OF THE INVENTION
`Tumor necrosis factor C, or TNFC, 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, cardio
`vascular effects, hemorrhage, coagulation, and acute phase
`responses Similar to those Seen during acute infections and
`Shock States. Excessive or unregulated TNFC. 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 (8.690), 662 (1990) and Adult Respiratory
`Distress Svindrome where TNFC. 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 TNFC. also
`resulted in changes typically seen in ARDS Ferrai
`Baliviera et al., Arch. Surg. 124(12), 1400–1405 (1989)}.
`TNFC. appears to be involved in bone resorption diseases,
`including arthritis. When activated, leukocytes will produce
`bone-resorption, an activity to which the data Suggest TNFC.
`contributes. Bertolini et al., Nature 319, 516–518 (1986)
`and Johnson et al., Endocrinology 124(3), 1424–1427
`(1989). TNFC. 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
`TNFC. may be involved in many bone resorption diseases,
`including arthritis, the most compelling link with disease is
`the association between production of TNFC. 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 TNFC. 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 Syn
`drome associated with high blood levels of TNFC. and the
`most Severe complication occurring in malaria patients.
`Levels of serum TNFC. 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)}.
`
`25
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
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`
`US 6,281.230 B1
`
`2
`Macrophage-induced angiogenesis TNFC. is known to be
`mediated by TNFC. Leibovich et al. Nature, 329, 630-632
`(1987) showed TNFC. induces in vivo capillary blood
`vessel formation in the rat cornea and the developing chick
`chorloallantoic membranes at very low doses and Suggest
`TNFC. is a candidate for inducing angiogenesis in
`inflammation, wound repair, and tumor growth. TNFC. pro
`duction 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).
`TNFC. 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 TNFC. completely
`blocked the silica-induced lung fibrosis in mice Pignet et
`al., Nature, 344:245–247 (1990). High levels of TNFC.
`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 TNFC, as compared with mac
`rophages from normal donors {Baughman et al., J. Lab.
`Clin. Med. 115(1), 36–42 (1990)}.
`TNFC. 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)}. TNFa. 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). TNFC. 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
`TNFC-induced expression of adhesion molecules, Such as
`intercellular adhesion molecule (ICAM) or endothelial leu
`kocyte adhesion molecule (ELAM) on endothelial cells
`{Munro et al., Am. J. Path. 135(1), 121-132 (1989).
`TNFC. blockage with monoclonal anti-TNFC. antibodies
`has been shown to be beneficial in rheumatoid arthritis
`{Elliot et al., Int. J. Pharmac. 1995 17(2), 141-145 and
`Crohn's disease von Dullemen et al., Gastroenterology,
`1995 109(1), 129-135}
`Moreover, it now is known that TNFC. is a potent activator
`of retrovirus replication including activation of HIV-1. Duh
`et al., Proc. Nat. Acad. Sci. 86,5974-5978 (1989); Pollet 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 lympho
`
`ALVOGEN, Exh. 1028, p. 0003
`
`
`
`US 6,281.230 B1
`
`4
`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. TNFC. and
`NFKB levels are influenced by a reciprocal feedback loop.
`AS noted above, the compounds of the present invention
`affect the levels of both TNFO and NFKB.
`Many cellular functions are mediated by levels of adenos
`ine 3',5'-cyclic monophosphate (cAMP). Such cellular func
`tions 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 TNFC. and
`NFKB. Increased levels of cAMP also leads to the relaxation
`of airway Smooth muscle.
`Decreasing TNFC. levels and/or increasing cAMP levels
`thus constitutes a valuable therapeutic Strategy for the treat
`ment of many inflammatory, infectious, immunological, and
`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, con
`gestive heart failure, fibrotic disease, cachexia, graft
`rejection, oncogenic or cancerous conditions, asthma,
`autoimmune disease, opportunistic infections in AIDS, rheu
`matoid arthritis, rheumatoid spondylitis, osteoarthritis, other
`arthritic conditions, Crohn's disease, ulcerative colitis, mul
`tiple Sclerosis, Systemic lupus erythrematosis, ENL in
`leprosy, radiation damage, oncogenic conditions, and hyper
`oxic alveolar injury. Prior efforts directed to the Suppression
`of the effects of TNFC. 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); WO92/11383}.
`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 TNFC.
`In particular, the invention pertains to (i) compounds of
`the formula:
`
`15
`
`25
`
`35
`
`40
`
`3
`cyte must continue to be maintained in an activated State to
`permit HIV gene expression and/or HIV replication.
`Cytokines, Specifically TNFC, are implicated in activated
`T-cell mediated HIV protein expression and/or virus repli
`cation by playing a role in maintaining T lymphocyte
`activation. Therefore, interference with cytokine activity
`Such as by prevention or inhibition of cytokine production,
`notably TNFC, 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 main
`tenance 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. {Rosen
`berg et al., The Immunopathogenesis of HIV Infection,
`Advances in Immunology, 57 (1989). Cytokines, such as
`TNFC, 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 iden
`tified TNFO 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 sug
`gests that a reduction of TNFC. synthesis may have an
`antiviral effect in HIV infections, by reducing the transcrip
`tion and thus virus production.
`AIDS viral replication of latent HIV in T cell and mac
`rophage lines can be induced by TNFC. Folksel al., PNAS
`86, 2365-2368 (1989). A molecular mechanism for the
`virus inducing activity is Suggested by TNFC.'s ability to
`activate a gene regulatory protein (NFKB) 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). TNFC. in AIDS asso
`ciated cachexia is Suggested by elevated Serum TNFC. and
`high levels of spontaneous TNFC. production in peripheral
`blood monocytes from patients Wright et al., J. Immunol.
`141(1), 99-104 (1988). TNFC. has been implicated in
`various roles with other viral infections, Such as the cytome
`galia virus (CMV), influenza virus, adenovirus, and the
`herpes family of Viruses for Similar reasons as those noted.
`The nuclear factor KB (NFKB) is a pleiotropic transcrip
`tional activator (Lenardo, et al., Cell 1989, 58, 227-29).
`NFKB 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 TNFC.
`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 el al., Bio
`chem. 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 NFkB 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 NFkB 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
`
`45
`
`50
`
`55
`
`60
`
`65
`
`R2
`
`R3
`
`R1
`
`R4
`
`X R'
`N
`N
`M
`Y
`
`O
`
`NH
`
`O
`
`in which:
`one of X and Y is C=O and the other of X and Y is C=O
`or CH;
`(i) each of R', R, R, and R", independently of the others,
`is halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to
`4 carbon atoms or (ii) one of R', R, R, and R' is
`-NHR and the remaining of R', R, R, and R' are
`hydrogen;
`R is hydrogen or alkyl of 1 to 8 carbon atoms;
`R is hydrogen, alkyl of 1 to 8 carbon atoms, benzyl, or
`halo,
`provided that R is other than hydrogen if X and Y are
`C=O and (i) each of R', R, R, and R' is fluoro or (ii)
`one of R', R, R, or R' is amino; and
`
`ALVOGEN, Exh. 1028, p. 0004
`
`
`
`US 6,281.230 B1
`
`6
`-continued
`O
`
`CIH.N.
`
`H
`
`S
`(b) the acid addition salts of Said compounds which contain
`a nitrogen atom capable of being protonated.
`A preferred group of compounds are those of Formula I in
`which each of R', R, R, and R", independently of the
`others, is halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1
`to 4 carbon atoms, and R is hydrogen, methyl, ethyl, or
`propyl. A Second preferred group of compounds are those of
`Formula I in which one of R', R, R, and R' is -NH, the
`remaining of R', R, R, and R" are hydrogen, and R is
`hydrogen, methyl, ethyl, or propyl.
`Unless otherwise defined, the term alkyl denotes a uni
`Valent 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. Preferably R', R,
`R, and R' are chloro, fluoro, methyl or methoxy.
`The compounds of Formula I are used, under the Super
`Vision of qualified professionals, to inhibit the undesirable
`effects of TNFC. 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 TNFC.
`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 treat
`ment of mammals other than humans in need of prevention
`or inhibition of TNFC. production. TNFC. mediated diseases
`for treatment, therapeutically or prophylactically, in animals
`include disease States Such as those noted above, but in
`40
`particular viral infections. Examples include feline immu
`nodeficiency virus, equine infectious anaemia virus, caprine
`arthritis virus, Visna Virus, and maedi Virus, as well as other
`lentiviruses.
`Compounds in which one of R', R, R, R" is amino and
`R and R, as well as the remainder of R', R, R, R', are
`hydrogen, as for example, 1,3-dioxo-2-(2,6-dioxopiperidin
`3-yl)-4-aminoiso indoline or 1,3-dioxo-2-(2,6-
`dioxopiperidin-3-yl)-5-aminoisoindoline are known. See,
`e.g. Jónsson, Acta Pharma. Succica, 9,521-542 (1972).
`The compounds can be prepared using methods which are
`known in general. In particular, the compounds can be
`prepared through the reaction of 2,6-dioxopiperidin-3-
`ammonium chloride, and a lower alkyl ester of
`2-bromomethylbenzoic acid in the presence of an acid
`acceptor Such as dimethylaminopyridine or triethylamine.
`
`The Substituted benzoate intermediates are known or can
`be obtained though conventional processes. For example, a
`lower alkyl ester of an ortho-toluic acid is brominated with
`N-bromosuccinimide under the influence of light to yield the
`lower alkyl 2-bromomethylbenzoate.
`
`Alternatively, a dialdehyde is allowed to react with 2,6-
`dioxopiperidin-3-ammonium chloride:
`
`R1
`
`CHO
`
`CHO
`
`In a further method, a dialdehyde is allowed to react with
`glutamine and the resulting 2-(1-oxoisoindoiln-2-yl)glutaric
`acid then cyclized to yield a 1-oxo-2-(2,6-dioxopiperidin-3-
`yl)-isoindoline of Formula I:
`
`R1
`
`R2
`
`CHO
`
`--
`
`R3
`
`CHO
`
`R4
`
`CHN
`
`COOH
`
`R6
`
`He
`
`CONH2
`
`1.
`R
`
`O
`
`R2
`
`I -H
`
`R
`
`N
`
`COOH
`
`R3
`
`R4
`
`CONH2
`
`Finally, an appropriately Substituted phthalidimide inter
`mediate is Selectively reduced:
`
`1O
`
`15
`
`25
`
`35
`
`45
`
`50
`
`55
`
`COOalkyl
`
`CHBr
`
`60
`
`65
`
`ALVOGEN, Exh. 1028, p. 0005
`
`
`
`7
`
`1.
`R
`
`O
`
`O
`
`R
`
`N
`
`R4
`
`O
`
`R2
`
`R3
`
`H2
`Her I
`
`5
`
`H Y
`N
`
`O
`
`Amino compounds can be prepared through catalytic
`hydrogenation of the corresponding nitro compound:
`
`O
`
`15
`
`X R.
`M
`N
`M
`Y
`
`H
`N1
`
`O
`
`ON
`
`The nitro intermediates of Formula IA are known or can
`be obtained though conventional processes. For example, a
`nitrophthalic anhydride is allowed to react with
`C-aminoglutarimide hydrochloride alternatively named as
`2,6-dioxopiperidin-3-ylarmmonium chloride in the pres
`ence of Sodium acetate and glacial acetic acid to yield an
`intermediate of Formula IA in which X and Y are both
`C=O.
`In a Second route, a lower alkyl ester of nitro-ortho-toluic
`acid is brominated with N-bromosuccinimide under the
`influence of light to yield a lower alkyl 2-(bromomethyl)
`nitrobenzoate. This is allowed to react with 2,6-
`dioxopiperidin-3-ammonium chloride in, for example, dim
`ethylformamide in the presence of triethylamine to yield an
`intermediate of Formula II in which one of X is C=O and
`the other is CH.
`Alternatively, if one of R, R2, R, and R is protected
`amino, the protecting group can be cleaved to yield the
`corresponding compound in which one of R, R2, R, and R.
`is amino. 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 protect
`ing groups thus are of importance primarily as chemical
`intermediates (although Some derivatives also exhibit bio
`logical 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”, Ple
`num Press, London and N.Y., 1973; Greene, Th. W. “Pro
`tective Groups in Organic Synthesis”, Wiley, New York,
`1981; “The Peptides”, Vol. I, Schröder and Lubke, Academic
`Press, London and New York, 1965; “Methoden der orga
`nischen Chemie', Houben-Weyl, 4th Edition. Vol.15/I,
`Georg Thieme Verlag, Stuttgart 1974, the disclosures of
`which are incorporated herein by reference. 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 C. position to the carbonyl group,
`particularly tertiary alkanoyl Such as pivaloyl, a lower
`alkanoyl group which is Substituted in the position C. to the
`carbonyl group, as for example trifluoroacetyl.
`
`25
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`US 6,281.230 B1
`
`8
`The compounds of the present invention possess a center
`of chirality 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 present invention.
`The racemates can be used as Such or can be separated into
`their individual isomers mechanically as by chromatography
`using a chiral adsorbent. Alternatively, the individual iso
`mers can be prepared in chiral form or separated chemically
`from a mixture by forming Salts with a chiral acid, Such as
`the individual enantiomers of 10-camphorSulfonic acid,
`camphoric acid, C.-bromocamphoric acid, methoxyacetic
`acid, tartaric acid, diacetyltartaric acid, malic acid,
`pyrrollidone-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 compounds 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, maleic acid, maleic acid, Sorbic
`acid, aconitic acid, Salicylic acid, phthalic acid, embonic
`acid, enanthic acid, and the like.
`Oral dosage forms include tablets, capsules, dragees, and
`Similar shaped, compressed pharmaceutical forms contain
`ing 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 admin
`istration. 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 the present invention associated with at least
`one pharmaceutically acceptable carrier, diluent or excipi
`ent. 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 Solu
`tions and Sterile packaged powders. Examples of Suitable
`excipients include lactose, dextrose, Sucrose, Sorbitol,
`mannitol, Starch, gum acacia, calcium Silicate, microcrys
`talline cellulose, polyvinylpyrrolidinone, cellulose, water,
`Syrup, and methyl cellulose, the formulations can addition
`ally 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 following examples will serve to further typify the
`nature of this invention but should not be construed as a
`limitation in the Scope thereof, which Scope is defined Solely
`by the appended claims.
`EXAMPLE 1.
`1,3-Dioxo-2-(2,6-dioxopiperidin-3-yl)-5-
`aminoisoindoline
`A mixture of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-
`nitroisoindoline alternatively named as N-(2.6-
`
`ALVOGEN, Exh. 1028, p. 0006
`
`
`
`US 6,281.230 B1
`
`9
`dioxopiperidin-3-yl)-4-nitrophthalimide (1 g, 3.3 mmol)
`and 10% Pd/C (0.13 g) in 1,4-dioxane (200 mL) was
`hydrogenated at 50 psi for 6.5 hours. The catalyst was
`filtered through Celite and the filtrate concentrated in vacuo.
`The residue was crystallized from ethyl acetate (20 mL) to
`give 0.62 g (69%) of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-
`5-aminoisoindoline alternatively named as N-(2,6-
`dioxopiperidin-3-yl)-4-aminophthalimide as an orange
`Solid. Recrystallization from dioxane/ethyl acetate gave 0.32
`g of yellow solid: mp 318.5–320.5° C.: HPLC (nova Pak
`C18.15/85 acetonitrile/0.1% HPO)3.97 min (98.22%); H
`NMR (DMSO-d) 811.08(s, 1H), 7.53–7.50 (d. J=8.3 Hz,
`1H), 6.94(s, 1H). 6.84–6.81(d, J=8.3 Hz,1H), 6.55(s.2H).
`5.05–4.98(m, 1H), 2.87–199(m, 4H); C NMR (DMSO
`d) 8.172.79, 170.16. 167.65, 167.14, 155.23, 134.21, 125.22
`15
`116.92, 116.17, 107.05, 48.58, 30.97, 22.22; Anal. Calcd for
`CHNO: C, 57.14; H, 4.06; N, 15.38. Found: C, 56.52
`H, 4.17; N, 14.60.
`In a similar fashion from 1-oxo-2-(2,6-dioxopiperidin-3-
`yl)-5-nitroisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-
`nitroisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-6-
`nitroisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-7-
`nitroisoindoline, and 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-
`4-nitroisoindoline, there is respectively obtained 1-oxo-2-
`(2,6-dioxopiperidin-3 -yl)-5-aminoisoindoline, 1-oxo-2-(2,
`6-dioxopiperidin-3-yl)-4-aminoisoindoline, 1-oxo-2-(2,6-
`dioxopiperidin-3-yl)-6-aminoisoindoline, 1-oxo-2-(2,6-
`dioxopiperidin-3-yl)-7-aminoisoindoline, and 1,3-dioxo-2-
`(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, respectively,
`upon hydrogenation.
`
`25
`
`10
`conventional bromination with N-bromosuccinimide under
`the influence of light.
`
`EXAMPLE 3
`1-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-
`tetrafluoroi