(12) United States Patent
`Muller et al.
`
`US006281230B1
`US 6,281,230 B1
`Aug. 28, 2001
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`(54)
`
`(75)
`
`(73)
`(*)
`
`ISOINDOLINES, METHOD OF USE, AND
`PHARMACEUTICAL COMPOSITIONS
`
`Inventors: George W. Muller, Bridgewater; David
`I. Stirling, Branchburg; Roger
`Shen-Chu Chen, Edison, all of NJ
`(Us)
`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)
`(22)
`
`Appl. No.: 09/543,809
`Filed:
`Apr. 6, 2000
`
`(62)
`
`(60)
`
`(51)
`
`(52)
`
`(58)
`
`(56)
`
`Related US. Application Data
`
`Division of application No. 09/230,389, ?led as application
`No. PCT/US97/13375 on Jul. 24, 1997, now abandoned,
`which is a continuation of application No. 08/690,258, ?led
`on Aug. 22, 1996, now Pat. No. 5,798,368, which is a
`continuation of application No. 08/701,499, ?led on Jul. 24,
`1996, now Pat. No. 5,635,517
`Provisional application No. 60/048,278, ?led on May 30,
`1997.
`
`Int. Cl.7 ..................... .. A61K 31/445; C07D 401/04
`
`US. Cl. ........................................... .. 514/323; 546/201
`
`Field of Search ............................ .. 514/323; 546/201
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`5/1986 Hiraga et al. .
`4,590,189
`2/1989 Leibovich et al. .
`4,808,402
`7/1989 OkaZaki et al. .
`4,849,441
`1/1995 Kaplan et al. .
`5,385,901
`10/1995 Muller et al. .
`5,463,063
`3/1996 Heinemann et al. .
`5,502,066
`1/1997 D’Amato.
`5,593,990
`2/1997 Muller.
`5,605,914
`5/1997 D’Amato.
`5,629,327
`5,635,517 * 6/1997 Muller et al. ...................... .. 514/323
`5,658,940
`8/1997 Muller et al. .
`5,698,579
`12/1997 Muller.
`5,703,098
`12/1997 Muller et al. .
`5,712,291
`1/1998 D’Amato.
`5,728,845
`3/1998 Muller et al. .
`5,736,570
`4/1998 Muller et al. .
`5,798,368
`8/1998 Muller et al. .
`5,801,195
`9/1998 Muller et al. .
`5,874,448 * 2/1999 Muller et al. ...................... .. 514/323
`5,877,200
`3/1999 Muller.
`5,929,117
`7/1999 Muller et al. .
`5,968,945
`10/1999 Muller et al. .
`6,011,050
`1/2000 Muller et al. .
`6,020,358
`2/2000 Muller et al. .
`6,046,221
`4/2000 Muller et al. .
`6,071,948
`6/2000 D’Amato.
`
`FOREIGN PATENT DOCUMENTS
`
`95/01348
`
`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 Con?guration of the Enantiomers of
`7—Chloro—4—[[4—diethylamino)—1—methylbuty]amino]
`quinoline (chloroquine)”, J. Org. Chem., vol. 53, pp.
`1167—1170, 1988.
`Feiser, Louis E, Experiments in Organic Chemistry, 3rd
`edition, p. 75, 1955.
`He, Weixuan et al., “Synthesis of Thalidomide Analogs and
`Their Biological Potential for Treatment of Graft Versus
`Host 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—In?amma
`tory Agents, Progress in Medicinal Chemistry, vol. 22, pp.
`166—242 (1985).
`Miyachi, H. et al., Novel Biological Response Modi?ers:
`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:
`In?uence 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 Tetra?uorothalidomide and Tetr
`l?uorophtalimides, J. Med. Chem., pp. 3044—3045 (1996).
`Smith, R. L. et al., Studies on the Relationship Between the
`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
`ABSTRACT
`(57)
`Substituted 1-oxo-2-(2,6-dioxopiperidin-3-yl)isoindolines
`are useful in treating in?ammation, in?ammatory 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
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 1
`
`

`

`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 Modi?cations 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 Per?uori
`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 In?ammation, pp. 177—211
`(1996).
`
`* cited by examiner
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 2
`
`

`

`US 6,281,230 B1
`
`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 ?led Jul.
`24, 1997, Which is a continuation of Ser. No. 08/690,258
`?led Jul. 24, 1996, now US. Pat. No. 5,635,517, Which is a
`continuation of Ser. No. 08/701,494 ?led Aug. 22, 1996 now
`US. Pat. No. 5,798,368, and provisional application Ser.
`No. 60/048,278 ?led 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 0t in a mammal
`through the administration thereof, and pharmaceutical com
`positions of such derivatives.
`
`BACKGROUND OF THE INVENTION
`
`Tumor necrosis factor 0t, or TNFot, is a cytokine Which is
`released primarily by mononuclear phagocytes in response
`to a number immunostimulators. When administered to
`animals or humans, it causes in?ammation, fever, cardio
`vascular effects, hemorrhage, coagulation, and acute phase
`responses similar to those seen during acute infections and
`shock states. Excessive or unregulated TNFO. 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 Svndrome Where TNFO. 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 TNFO. also
`resulted in changes typically seen in ARDS {Ferrai
`Baliviera et al.,Arch. Surg. 124(12), 1400—1405 (1989)}.
`TNFO. appears to be involved in bone resorption diseases,
`including arthritis. When activated, leukocytes Will produce
`bone-resorption, an activity to Which the data suggest TNFO.
`contributes. {Bertolini et al., Nature 319, 516—518 (1986)
`and Johnson et al., Endocrinology 124(3), 1424—1427
`(1989).} TNFO. 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
`TNFO. may be involved in many bone resorption diseases,
`including arthritis, the most compelling link With disease is
`the association betWeen production of TNFO. 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 TNFO. 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 TNFO. and the
`most severe complication occurring in malaria patients.
`Levels of serum TNFO. 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)}.
`
`10
`
`15
`
`25
`
`35
`
`45
`
`55
`
`65
`
`2
`Macrophage-induced angiogenesis TNFO. is knoWn to be
`mediated by TNFot. Leibovich et al. {Nature, 329, 630—632
`(1987)} shoWed TNFO. induces in vivo capillary blood
`vessel formation in the rat cornea and the developing chick
`chorloallantoic membranes at very loW doses and suggest
`TNFO. is a candidate for inducing angiogenesis in
`in?ammation, Wound repair, and tumor groWth. TNFO. 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)}.
`TNFO. also plays a role in the area of chronic pulmonary
`in?ammatory diseases. The deposition of silica particles
`leads to silicosis, a disease of progressive respiratory failure
`caused by a ?brotic reaction. Antibody to TNFO. completely
`blocked the silica-induced lung ?brosis in mice {Pignet et
`al., Nature, 344:245—247 (1990)}. High levels of TNFO.
`production (in the serum and in isolated macrophages) have
`been demonstrated in animal models of silica and asbestos
`induced ?brosis {Bissonnette et al., In?ammation 13(3),
`329—339 (1989)}. Alveolar macrophages from pulmonary
`sarcoidosis patients have also been found to spontaneously
`release massive quantities of TNFO. as compared With mac
`rophages from normal donors {Baughman et al., J. Lab.
`Clin. Med. 115(1), 36—42 (1990)}.
`TNFO. is also implicated in the in?ammatory response
`Which folloWs reperfusion, called reperfusion injury, and is
`a major cause of tissue damage after loss of blood ?oW
`{Vedder et al., PNAS 87, 2643—2646 (1990)}. TNFO. 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)}. TNFO. has pro-in?ammatory
`activities Which together With its early production (during
`the initial stage of an in?ammatory 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 speci?c importance may be
`TNFot-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)}.
`TNFO. blockage With monoclonal anti-TNFot antibodies
`has been shoWn to be bene?cial 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 TNFO. 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 Immunode?ciency Virus (HIV).
`At least three types or strains of HIV have been identi?ed,
`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
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 3
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`

`

`US 6,281,230 B1
`
`3
`cyte must continue to be maintained in an activated state to
`permit HIV gene expression and/or HIV replication.
`Cytokines, speci?cally TNFot, 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 TNFot, 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
`TNFot, 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
`ti?ed 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 TNFO. 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 TNFO. {Folks el al., PNAS
`86, 2365—2368 (1989)}. A molecular mechanism for the
`virus inducing activity is suggested by TNFot’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)}. TNFO. in AIDS asso
`ciated cacheXia is suggested by elevated serum TNFO. and
`high levels of spontaneous TNFO. production in peripheral
`blood monocytes from patients {Wright et al., J. Immunol.
`141(1), 99—104 (1988)}. TNFO. has been implicated in
`various roles With other viral infections, such as the cytome
`galia virus (CMV), in?uenza 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 in?ammatory states and is thought to
`regulate cytokine levels including but not limited to TNFO.
`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
`De?ciency Syndrome 1993, 6, 778—786; Suzuki el al., Bio
`chem. And Biophys. Res. Comm. 1993, 193, 277—83; SuZuki
`et al., Biochem. Ana' 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
`
`15
`
`25
`
`35
`
`45
`
`55
`
`65
`
`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. TNFO. and
`NFKB levels are in?uenced 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 in?ammatory conditions and diseases
`including asthma, in?ammation, 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 in?ammatory
`leukocytes inhibits their activation and the subsequent
`release of in?ammatory mediators, including TNFO. and
`NFKB. Increased levels of cAMP also leads to the relaxation
`of airWay smooth muscle.
`Decreasing TNFO. levels and/or increasing cAMP levels
`thus constitutes a valuable therapeutic strategy for the treat
`ment of many in?ammatory, 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, ?brotic 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 TNFO. 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}.
`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 TNFot.
`In particular, the invention pertains to
`compounds of
`the formula:
`
`R2
`
`R3
`
`R1
`
`R4
`
`X R6
`\
`N
`/
`Y
`
`0
`
`NH
`
`0
`
`in Which:
`one of X andY is C=O and the other of X and Y is C=O
`or CH2;
`(i) each of R1, R2, R3, and R4, 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 R1, R2, R3, and R4 is
`—NHR5 and the remaining of R1, R2, R3, and R4 are
`hydrogen;
`R5 is hydrogen or alkyl of 1 to 8 carbon atoms;
`R6 is hydrogen, alkyl of 1 to 8 carbon atoms, benZyl, or
`halo;
`provided that R6 is other than hydrogen if X and Y are
`C=O and
`each of R1, R2, R3, and R4 is ?uoro or (ii)
`one of R1, R2, R3, or R4 is amino; and
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 4
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`

`US 6,281,230 B1
`
`5
`(b) the acid addition salts of said compounds Which contain
`a nitrogen atom capable of being protonated.
`Apreferred group of compounds are those of Formula I in
`Which each of R1, R2, R3, and R4, independently of the
`others, is halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1
`to 4 carbon atoms, and R6 is hydrogen, methyl, ethyl, or
`propyl. Asecond preferred group of compounds are those of
`Formula I in Which one of R1, R2, R3, and R4 is —NH2, the
`remaining of R1, R2, R3, and R4 are hydrogen, and R6 is
`hydrogen, methyl, ethyl, or propyl.
`Unless otherWise de?ned, 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 R1, R2,
`R3, and R4 are chloro, ?uoro, methyl or methoxy.
`The compounds of Formula I are used, under the super
`vision of quali?ed professionals, to inhibit the undesirable
`effects of TNFot. 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 TNFO.
`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 TNFO. production. TNFO. 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 immu
`node?ciency virus, equine infectious anaemia virus, caprine
`arthritis virus, visna virus, and maedi virus, as Well as other
`lentiviruses.
`Compounds in Which one of R1, R2, R3, R4 is amino and
`R5 and R6, as Well as the remainder of R1, R2, R3, R4, are
`hydrogen, as for example, 1,3-dioxo-2-(2,6-dioxopiperidin
`3-yl)-4-aminoisoindoline or 1,3-dioxo-2-(2,6
`dioxopiperidin-3-yl)-5-aminoisoindoline are knoWn. See,
`eg Jonsson, 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.
`
`COOalkyl
`
`R3
`
`CHZBr
`
`6
`-continued
`0
`
`1O
`
`15
`
`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 in?uence of light to yield the
`loWer alkyl 2-bromomethylbenZoate.
`
`Alternatively, a dialdehyde is alloWed to react With 2,6
`dioxopiperidin-3-ammonium chloride:
`
`R1
`
`R2
`
`CH0
`
`25
`
`CH0
`
`35
`
`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
`
`45
`
`R3
`
`+
`
`CHO
`
`R4
`
`C1'H3N"
`
`COOH
`
`R6
`
`—>
`
`55
`
`65
`
`CONHZ
`
`1
`R
`
`o
`
`R2
`
`I <—
`
`R6
`
`N
`
`COOH
`
`R3
`
`R4
`
`CONHZ
`
`Finally, an appropriately substituted phthalidimide inter
`mediate is selectively reduced:
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 5
`
`

`

`US 6,281,230 B1
`
`7
`
`1
`R
`
`O
`
`O
`
`R6
`
`N
`
`R4
`
`0
`
`R2
`
`R3
`
`H
`/
`N
`
`0
`
`H2
`—> I
`
`Amino compounds can be prepared through catalytic
`hydrogenation of the corresponding nitro compound:
`
`0
`
`15
`
`X R6
`\
`N
`/
`Y
`
`H
`N/
`
`O
`
`OZN
`
`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
`ot-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
`in?uence 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 CH2.
`Alternatively, if one of R1, R2, R3, and R4 is protected
`amino, the protecting group can be cleaved to yield the
`corresponding compound in Which one of R1, R2, R3, and R4
`is amino. Protecting groups utiliZed herein denote groups
`Which generally are not found in the ?nal 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 NY, 1973; Greene, Th. W. “Pro
`tective Groups in Organic Synthesis”, Wiley, NeW York,
`1981; “The Peptides”, Vol. I, Schroder 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 0t position to the carbonyl group,
`particularly tertiary alkanoyl such as pivaloyl, a loWer
`alkanoyl group Which is substituted in the position 0t to the
`carbonyl group, as for example tri?uoroacetyl.
`
`25
`
`35
`
`45
`
`55
`
`65
`
`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, ot-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 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 ?avoring
`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 de?ned 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
`
`DR. REDDY’S LABS., INC. EX. 1006 PAGE 6
`
`

`

`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
`?ltered through Celite and the ?ltrate 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% H3PO4) 3.97 min (98.22%): 1H
`NMR (DMSO-dG) 611.08(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—1.99(m, 4H); 13C NMR (DMSO
`d6) 6172.79, 170.16.167.65, 167.14, 155.23, 134.21, 125.22
`116.92, 116.17, 107.05, 48.58, 30.97, 22.22; Anal. Calcd for
`C13H11N3O4: 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