United States Patent [19]
`Muller et al.
`
`I II IIII Ill lllll lllll lllll lllll lllll lllll lllll lllll 111111111111111111
`US005635517 A
`[111 Patent Number:
`[45] Date of Patent:
`
`5,635,517
`Jun. 3, 1997
`
`[54] METHOD OF REDUCING TNFa LEVELS
`WITH AMINO SUBSTITUTED 2-(2,6-
`DIOXOPIPERIDIN-3-YL)-1-0XO-AND 1,3-
`DIOXOISOINDOLINES
`
`[75]
`
`Inventors: George W. Muller, Bridgewater; David
`I. Stirling, Branchburg; Roger S. -C.
`Chen, Edison, all of N.J.
`
`[73] Assignee: Celgene Corporation. Warren, N.J.
`
`[21] Appl. No.: 690,258
`Jul. 24, 1996
`[22] Filed:
`Int. CI.6
`.....................•. A61K 31/445; C07D 401/04
`[51]
`[52] U.S. CI •............................................. 514/323; 546/201
`[58] Field of Search ............................. 546/201; 514/323,
`514/231.5, 231.2, 327
`
`[56]
`
`References Cited
`
`U.S. PATENT DOCUMENfS
`
`1/1995 Kaplan et al ........................ 514/231.5
`5,385,901
`5,463,083 10/1995 Muller ..................................... 546/201
`
`OTHER PUBLICATIONS
`
`N. Ake Jonsson, Chemical Structure and Teratogenic Prop(cid:173)
`erties: A Review Of Available Data On Structure-Activity
`Relationships and Mechanism Of Action Of Thalidomide
`
`Analogues, Acta Pharm. Suicica, vol. 9, pp. 521-542
`(1972).
`H. Koch, 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).
`L. Sekut et al., Pathophysiology and Regulation Of TNF-a
`In Inflammation, DN&P 9(5), pp. 261-269, (1996).
`Smith, R.L.; Fabro, S.; Schumacher, H. and William, R.T.;
`Studies on the relationship between the Chemical Structured
`and Embryotoxic Activity Of Thalidomide and Related
`Compounds. A Symposium on Embryotoxic Activity of
`dough, edited by Pobinson, J.M. et al. 1965, pp. 194-209.
`See p. 202, Table 6.
`
`Primary Examiner-C. Warren Ivy
`Assistant Examiner-C. S. Aulakh
`Attorney, Agent, or Firm-Mathews, Collins, Shepherd &
`Gould, P.A.
`
`[57]
`
`ABSTRACT
`
`1-0xo- and l,3-dioxo-2-(2,6-dioxopiperidin-3-yl) isoindo(cid:173)
`lines substituted with amino in the benzo ring reduce the
`levels of TNFa in a mammal. A typical embodiment is
`1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline.
`
`10 Claims, No Drawings
`
`DR. REDDY’S LABS., INC. EX. 1024 PAGE 1
`
`

`

`5,635,517
`
`1
`METHOD OF REDUCING TNFa LEVELS
`WITH AMINO SUBSTITUTED 2-(2,6-
`DIOXOPIPERIDIN-3-YL)-1-0XO-AND 1,3-
`DIOXOISOINDOLINES
`
`5
`
`BACKGROUND OF THE INVENTION
`
`The present invention relates a method of reducing levels
`of tumor necrosis factor a in a mammal through the admin(cid:173)
`istration of an amino substituted 2-(2,6-dioxopiperidin-3-yl)
`-1-oxoisoindolines and 1,3-dioxoisoindolines and to phar(cid:173)
`maceutical compositions of such amino substituted indoline
`derivatives.
`
`2
`sarcoidosis patients have also been found to spontaneously
`release massive quantities of 1NFa as compared with mac(cid:173)
`rophages from normal donors {Baughman et al., J. Lab.
`Clin. Med. 115(1), 36-42 (1990)}.
`1NFa 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)}. 1NFa also
`alters the properties of endothelial cells and has various
`10 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)}.1NFa has pro-inflammatory
`Tumor necrosis factor a, or TNFa, is a cytokine which is 15 activities which together with its early production ( during
`the initial stage of an inflammatory event) make it a likely
`released primarily by mononuclear phagocytes in response
`to a number immunostimulators. When administered to
`mediator of tissue injury in several important disorders
`animals or humans, it causes inflammation, fever, cardio-
`including but not limited to, myocardial infarction, stroke
`vascular effects, hemorrhage, coagulation, and acute phase
`and circulatory shock. Of specific importance may be
`responses similar to those seen during acute infections and 20 TNFa-induced expression of adhesion molecules, such as
`shock states. Excessive or unregulated TNFa production
`intercellular adhesion molecule (ICAM) or endothelial leu-
`thus has been implicated in a number of disease conditions.
`kocyte adhesion molecule (ELAM) on endothelial cells
`{Munro et al., Am. J. Path. 135(1), 121-132 (1989)}.
`These include endotoxemia and/or toxic shock syndrome
`{Tracey et al., Nature 330, 662-664 ( 1987) and Hinshaw et
`Moreover, it now is known thatTNFa is a potent activator
`al., Circ. Shock 30, 279-292 (1990)}; cachexia {Dezube et 25 ofretrovirus replication including activation of HIV-1. {Duh
`u A d s · 86 5<J74--5978 (1989) p ll t al
`al., Lancet, 335 (8690), 662 (1990)} and Adult Respiratory
`al p
`., roe. ,.at. ca . ci.
`,
`; o e
`.,
`et
`Distress Syndrome where 1NFa concentration in excess of
`Proc. Nat. Acad. Sci. 87, 782-785 (1990); Monto et al.,
`Blood 79, 2670 (1990); Clouse et al., J. Immunol. 142,
`12,000 pg/mL have been detected in pulmonary aspirates
`from ARDS patients {Millar et al., Lancet 2(8665), 712-714
`431-438 (1989); Poll et al., AIDS Res. Hum. Retrovirus,
`(1989)}. Systemic infusion of recombinant TNFa also 30 191-197 (1992)}. AIDS results from the infection of T
`lymphocytes with Human Immunodeficiency Virus (HIV).
`resulted in changes typically seen in ARDS {Ferrai-
`Baliviera et al., Arch. Surg. 124(12), 1400-1405 (1989)}.
`At least three types or strains of HIV have been identified,
`TNFa appears to be involved in bone resorption diseases,
`i.e., HIV-1, HIV-2 and HIV-3. As a consequence of HIV
`including arthritis. When activated, leukocytes will produce
`infection, T-cell mediated immunity is impaired and infected
`bone-resorption, an activity to which the data suggest 1NFa 35 individuals manifest severe opportunistic infections and/or
`contributes. {Bertolini et al., Nature 319, 516-518 (1986)
`unusual neoplasms. HIV entry into the T lymphocyte
`and Johnson et al., Endocrinology 124(3), 1424--1427
`requires T lymphocyte activation. Other viruses, such as
`(1989).} TNFa also has been shown to stimulate bone
`HIV-1, HIV-2 infect T lymphocytes after T cell activation
`resorption and inhibit bone formation in vitro and in vivo
`and such virus protein expression and/or replication is
`through stimulation of osteoclast formation and activation 40 mediated or maintained by such T cell activation. Once an
`combined with inhibition of osteoblast function. Although
`activated T lymphocyte is infected with HIV, the T lympho-
`1NFa may be involved in many bone resorption diseases,
`cyte must continue to be maintained in an activated state to
`permit HIV gene expression and/or HIV replication.
`including arthritis, the most compelling link with disease is
`the association between production of 1NFa by tumor or
`Cytokines, specifically TNFa, are implicated in activated
`host tissues and malignancy associated hypercalcemia 45 T-cell mediated HIV protein expression and/or virus repli-
`{ Calci. Tissue Int. (US) 46(Suppl.), S3-10 (1990)}. In Graft
`cation by playing a role in maintaining T lymphocyte
`versus Host Reaction, increased serum TNFa levels have
`activation. Therefore, interference with cytokine activity
`been associated with major complication following acute
`such as by prevention or inhibition of cytokine production,
`allogenic bone marrow transplants {Holler et al., Blood,
`notably TNFa, in an HIV-infected individual assists in
`50 limiting the maintenance of T lymphocyte caused by HIV
`75(4), 1011-1016 (1990)}.
`Cerebral malaria is a lethal hyperacute neurological syn-
`infection.
`drome associated with high blood levels of TNFa and the
`Monocytes, macrophages, and related cells, such as
`most severe complication occurring in malaria patients.
`kup:ffer and glial cells, also have been implicated in main-
`Levels of serum TNFa correlated directly with the severity
`tenance of the HIV infection. These cells, like T cells, are
`of disease and the prognosis in patients with acute malaria 55 targets for viral replication and the level of viral replication
`attacks {Grau et al., N. Engl. J. Med. 320(24), 1586-1591
`is dependent upon the activation state of the cells. {Rosen-
`berg et al., The Immunopathogenesis of HIV Infection,
`(1989)}.
`TNFa also plays a role in the area of chronic pulmonary
`Advances in Immunology, 57 (1989)}. Cytokines, such as
`TNFa, have been shown to activate HIV replication in
`inflammatory diseases. The deposition of silica particles
`leads to silicosis, a disease of progressive respiratory failure 60 monocytes and/or macrophages {Poli et al. Proc. Natl. Acad.
`caused by a fibrotic reaction. Antibody to TNFa completely
`Sci., 87, 782-784 (1990)}, therefore, prevention or inhibi-
`tion of cytokine production or activity aids in limiting HIV
`blocked the silica-induced lung fibrosis in mice {Pignet et
`al., Nature, 344:245-247 (1990)}. High levels of 1NFa
`progression for T cells. Additional studies have identified
`TNFa as a common factor in the activation of HIV in vitro
`production (in the serum and in isolated macrophages) have
`been demonstrated in animal models of silica and asbestos 65 and has provided a clear mechanism of action via a nuclear
`regulatory protein found in the cytoplasm of cells (Osborn,
`induced fibrosis {Bissonnette et al., Inflammation 13(3),
`329-339 (1989)}. Alveolar macrophages from pulmonary
`et al., PNAS 86 2336-2340). This evidence suggests that a
`
`DR. REDDY’S LABS., INC. EX. 1024 PAGE 2
`
`

`

`5,635,517
`
`3
`reduction of TNFa. synthesis may have an antiviral effect in
`mv infections, by reducing the transcription and thus virus
`production.
`AIDS viral replication of latent mv in T cell and mac(cid:173)
`rophage lines can be induced by TNFa. {Folks et al., PNAS 5
`86, 2365-2368 (1989)}. A molecular mechanism for the
`virus inducing activity is suggested by TNFa.'s ability to
`activate a gene regulatory protein (NFJCB) found in the
`cytoplasm of cells, which promotes mv replication through
`binding to a viral regulatory gene sequence (LTR) {Osborn 10
`et al., PNAS 86, 2336-2340 (1989)}. TNFa. in AIDS asso(cid:173)
`ciated cachexia is suggested by elevated serum TNFa. and
`high levels of spontaneous TNFa. production in peripheral
`blood monocytes from patients {Wright et al. J. Immunol.
`141(1), 99-104 (1988)}. TNFa. has been implicated in 15
`various roles with other viral infections, such as the cytome(cid:173)
`galia virus (CMV), influenza virus, adenovirus, and the
`hetpes family of viruses for similar reasons as those noted.
`The nuclear factor 1CB (NFJCB) is a pleiotropic transcrip(cid:173)
`tional activator (Lenardo, et al. Cell 1989, 58, 227-29). 20
`NFJCB 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 TNFa.
`and also to be an activator of mv transcription (Dbaibo, et
`al. J. Biol. Chem. 1993, 17762-66; Duh et al. Proc. Natl. 25
`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; 30
`Suzuki et al. Biochem. Mal. 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 NFJCB binding can regulate
`transcription of cytokine gene(s) and through this modula- 35
`tion and other mechanisms be useful in the inhibition of a
`multitude of disease states. The compounds described herein
`can inhibit the action of NFJCB in the nucleus and thus are
`useful in the treatment of a variety of diseases including but
`not limited to rheumatoid arthritis, rheumatoid spondylitis, 40
`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, mv, AIDS, and opportu(cid:173)
`nistic infections in AIDS. TNFa. and NFJCB levels are 45
`influenced by a reciprocal feedback loop. As noted above,
`the compounds of the present invention affect the levels of
`both TNFa. and NFJCB.
`Many cellular functions are mediated by levels of adenos(cid:173)
`ine 3',5'-cyclic monophosphate ( cAMP). Such cellular func(cid:173)
`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 TNFa. and
`NFJCB. Increased levels of cAMP also leads to the relaxation
`of airway smooth muscle.
`Decreasing TNFa. levels and/or increasing cAMP levels
`thus constitutes a valuable therapeutic strategy for the treat- 60
`ment 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, con- 65
`gestive heart failure, fibrotic disease, cachexia, graft
`rejection, cancer, autoimmune disease, opportunistic infec-
`
`4
`tions 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 sup(cid:173)
`pression of the effects of TNFa. have ranged from the
`utilization of steroids such as dexamethasone and predniso(cid:173)
`lone to the use of both polyclonal and monoclonal antibodies
`{Beutler et al., Science 234, 470-474 (1985); WO
`92/11383}.
`
`DEfAILED DESCRIPTION
`
`The present invention is based on the discovery that a
`class of non-polypeptide compounds more fully described
`herein decrease the levels of TNFa. and elevate the levels of
`adenosine 3',5'-cyclic monophosphate.
`
`In particular, the invention pertains to the method of
`reducing undesirable levels of TNFa. in a mammal by
`administering to the mammal an effective amount of a
`compound of the formula:
`
`0 ~>6:
`
`H2N
`
`in which one of X and Y is C=O and the other of X and Y
`is C=O or CH2
`The compounds of Formula I are used, under the super(cid:173)
`vision of qualified professionals, to inhibit the undesirable
`effects ofTNFa.. 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 TNFa.
`production, respectively, such as viral infections, such as
`those caused by the hetpes viruses, or viral conjunctivitis,
`etc.
`The compounds also can be used in the veterinary treat(cid:173)
`ment of mammals other than humans in need of prevention
`50 or inhibition of TNFa. production. TNFa. 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-
`55 nodeficiency virus, equine infectious anaemia virus, caprine
`arthritis virus, visna virus, and maedi virus, as well as other
`lentiviruses.
`Certain of these compounds, such as 1,3-dioxo-2-(2,6-
`dioxopiperidin-3-yl)-4-aminoisoindoline and 1,3-dioxo-2-
`(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline are known.
`See, e.g., Jonsson, Acta Pharma. Succica, 9, 521-542
`(1972).
`In any event, the compounds can be prepared using
`methods which are known in general. In particular, the
`compounds can be prepared through catalytic hydrogenation
`of the corresponding nitro compound.
`
`DR. REDDY’S LABS., INC. EX. 1024 PAGE 3
`
`

`

`5,635,517
`
`5
`
`II
`
`6
`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
`5 compounds of Formula I associated with at least one phar(cid:173)
`maceutically 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 solu-
`15 tions and sterile packaged powders. Examples of suitable
`excipients include lactose, dextrose, sucrose, sorbitol,
`mannitol, starch, gum acacia, calcium silicate, microcrys(cid:173)
`talline cellulose, polyvinylpyrrolidinone, cellulose, water,
`syrup, and methyl cellulose, the formulations can addition-
`20 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 compositions preferably are formulated in unit dos-
`age 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 pro(cid:173)
`duce 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.
`Oral dosage forms include tablets, capsules, dragees, and
`similar shaped, compressed pharmaceutical forms contain(cid:173)
`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,
`40 intrathecal, intravenous and intra-arterial routes of admin(cid:173)
`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
`45 compounds of Formula I associated with at least one phar(cid:173)
`maceutically 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
`50 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
`55 hard gelatin capsules, suppositories, sterile injectable solu(cid:173)
`tions and sterile packaged powders. Examples of suitable
`excipients include lactose, dextrose, sucrose, sorbitol,
`mannitol, starch, gum acacia, calcium silicate, microcrys(cid:173)
`talline cellulose, polyvinylpyrrolidinone, cellulose, water,
`60 syrup, and methyl cellulose, the formulations can addition(cid:173)
`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
`65 agents.
`The compositions preferably are formulated in unit dos(cid:173)
`age form, meaning physically discrete units suitable as a
`
`30
`
`10
`
`The nitro intermediates of Formula II are known or can be
`obtained though conventional processes. For example, a
`nitrophthalic anhydride is allowed to react with
`a-arninoglutarimide hydrochloride { alternatively named as
`2,6-dioxopiperidin-3-ylammonium chloride} in the pres(cid:173)
`ence of sodium acetate and glacial acetic acid to yield an
`intermediate of Formula II 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(cid:173)
`ethylformarnide in the presence of triethylarnine to yield an
`intermediate of Formula II in which one of X is C=O and
`the other is CH2 •
`The compounds of Formula I 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 25
`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 absor(cid:173)
`bant Alternatively, the individual isomers 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,
`a-bromocamphoric acid, methoxyacetic acid, tartaric acid,
`diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic 35
`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%.
`Alternatively, the compounds can be stereopreferentially
`synthesized by allowing the lower alkyl 2-(bromomethyl)
`nitrobenzoate intermediate discussed above to react with
`either
`(R)-1- benzyloxy-2, 6-dioxo-3 -tert.(cid:173)
`butoxycarbonylarninopiperidine or (S)-l-benzyloxy-2,6-
`dioxo-3-tert-butoxycarbonylaminopiperidine analogous to
`the method described by Robin et al., Tetrahedron
`Asymmetry, 6, 1249 (1995). Hydrogenation in this case not
`only reduces the nitro group to an amino group but also
`converts the N-benzyloxy group to an N-hydroxy group
`which can be removed with bromoacetophenone triethy(cid:173)
`lamine and dimethylarninopyridine to yield the correspond(cid:173)
`ing (R) or (S) enantiomer of Formula L
`The present invention also pertains to the physiologically
`acceptable non-toxic acid addition salts of the compounds of
`Formula L 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.
`Oral dosage forms include tablets, capsules, dragees, and
`similar shaped, compressed pharmaceutical forms contain(cid:173)
`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-
`
`DR. REDDY’S LABS., INC. EX. 1024 PAGE 4
`
`

`

`5,635,517
`
`7
`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 pro(cid:173)
`duce the desired therapeutic effect in association with a 5
`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.
`Specific compounds falling within Formula I include 10
`l-oxo-2-(2,6-dioxopiperidin-3-yl)-5-arninoisoindoline,
`1-oxo-2-(2,6-dioxopiperidin-3-y 1)-4-arninoisoindoline,
`1-oxo-2-(2,6-dioxopiperidin-3-y 1 )-6-arninoisoindoline,
`1-oxo-2-(2,6-dioxopiperidin-3-yl)-7-arninoisoindoline, l,3-
`dioxo-2-(2,6-dioxopiperidin-3-yl)-4-arninoisoindoline, and 15
`1,3-dioxo-2-(2,6-dioxopiperidin-3 -yl)-5-arninoisoindoline.
`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 clainls.
`
`EXAMPLE 1
`
`8
`dioxopiperidin-3-yl)-5-nitroisoindoline as a light brown
`solid. An analytical sample was obtained by recrystallization
`from methanol: mp 228.5°-229.5° C.; 1H NMR (DMSO-d6)
`o ll.18(s, 1 H), 8.69-8.65(d,d J=l.9 and 8.0 Hz, lH),
`8.56(d, J=l.9 Hz, lH), 8.21(d, H=8.2 Hz, lH), 5.28(d,d
`1=5.3 and 12.8 Hz, lH), 2.93-2.07(m, 4H); 13C NMR
`(DMSO-d6) o 172.66, 169.47, 165.50, 165.23, 151.69,
`135.70, 132.50, 130.05, 124.97, 118.34, 49.46, 30.85, 21.79;
`Anal. Calcd for C 13IlgN30 6: C, 51.49; H, 2.99; N, 13.86.
`Found: C, 51.59; H, 3.07; N, 13.73.
`1-0xo-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-y 1)-6-nitroisoindoline, and
`1-oxo-2-(2,6-dioxopiperidin-3-yl)-7-nitroisoindoline can be
`obtained by allowing 2,6-dioxopiperidin-3-arnmonium chlo(cid:173)
`ride to react with methyl 2-bromomethyl-5-nitrobenzoate,
`methyl 2-bromomethyl-4-nitrobenzoate, methyl
`2-bromomethyl-6-nitrobenzoate, and methyl
`2-bromomethyl-7-nitrobenzoate, respectively, in dimethyl-
`20 formarnide in the presence of triethylarnine. The methyl
`2-(bromomethyl)nitrobenzoates in turn are obtained from
`the corresponding methyl esters of nitro-ortho-toluic acids
`by conventional brornination with N-bromosuccinirnide
`under the influence of light.
`
`A mixture of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-
`nitroisoindoline {alternatively named as N-(2,6-
`dioxopiperidin-3-yl)-4-nitrophthalinride} (1 g, 3.3 mmol) 25
`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-arninoisoindoline { alternatively named as N-(2,6-
`dioxopiperidin-3-yl)-4-arninophthalirnide} 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
`Cl8,15/85 acetonitrile/0.1%H3P04 ) 3.fJl min (98.22%); 1H 35
`NMR (DMSO-d6) o ll.08(s, lH), 7.53-7.50 (d, J=8.3 Hz,
`lH), 6.94(s, lH), 6.84-6.8l(d, J=8.3 Hz, lH), 6.55(s,2H).
`5.05-4.98(m, lH), 2.87-l.99(m, 4H); 13C NMR (DMSO-
`d6) o 172.79,170.16, 167.65, 167.14, 155.23, 134.21,
`125.22, 116.92, 116.17, 107.05, 48.58, 30.97, 22.22; Anal. 40
`Calcd for C13H 11N30 4 : 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-arninoisoindoline, 1-oxo-2-(2,
`6-dioxopiperidin-3-yl)-4-arninoisoindoline, 1-oxo-2-(2,6-
`dioxopiperidin-3-yl)-6-arninoisoindoline, 1-oxo-2-(2,6-
`dioxopiperidin-3-yl)-7-aminoisoindoline, and l,3-dioxo-2-
`(2,6-dioxopiperidin-3-yl)-4-arninoisoindoline, respectively,
`upon hydrogenation.
`
`EXAMPLE3
`
`Tablets, each containing 50 mg of 1,3-dioxo-2-(2,6-
`dioxopiperidin-3-yl)-5-aminoisoindoline, can be prepared in
`30 the following manner:
`
`Constituents (for 1000 tablets)
`
`1,3-dioxo-2-(2,6-dioxo(cid:173)
`piperidin-3-yl)-5-amino(cid:173)
`isoindoline
`lactose
`wheat starch
`polyethylene glycol 6000
`talc
`magnesium stearate
`demineralized water
`
`50.0 g
`
`50.7 g
`7.5 g
`5.0 g
`5.0 g
`1.8 g
`q.s.
`
`The solid ingredients are first forced through a sieve of 0.6
`mm mesh width. The active ingredient, lactose, talc, mag-
`45 nesium stearate and half of the starch then are mixed. The
`other half of the starch is suspended in 40 mL of water and
`this suspension is added to a boiling solution of the poly(cid:173)
`ethylene glycol in 100 rnL of water. The resulting paste is
`added to the pulverulent substances and the mixture is
`50 granulated, if necessary with the addition of water. The
`granulate is dried overnight at 35° C., forced through a sieve
`of 1.2 mm mesh width and compressed to form tablets of
`approximately 6 mm diameter which are concave on both
`sides.
`
`55
`
`EXAMPLB2
`
`EXAMPLE4
`
`Tablets, each containing 100 mg of 1,3-dioxo-2-(2,6-
`dioxopiperidin-3-yl)-5-aminoisoindoline, can be prepared in
`60 the following manner:
`
`A mixture of 4-nitrophthalic anhydride (1.7 g, 8.5 mmol),
`a-aminoglutarirnide hydrochloride (1.4 g, 8.5 mmol) and
`sodium acetate (0.7 g, 8.6 mmol) in glacial acetic acid (30
`mL) was heated under reflux for 17 hours. The mixture was
`concentrated in vacuo and the residue was stirred with
`methylene chloride (40 mL) and water (30 mL). The aque(cid:173)
`ous layer was separated, extracted with methylene chloride
`(2x40 mL). The combined methylene chloride solutions 65
`were dried over magnesium sulfate and concentrated in
`vacuo to give 1.4 g (54%) of l,3-dioxo-2-(2,6-
`
`Constituents (for 1000 tablets)
`
`l,3-dioxo-2-(2,6-dioxo(cid:173)
`piperidin-3-yl)-5-amino(cid:173)
`isoindoline
`
`100.0 g
`
`DR. REDDY’S LABS., INC. EX. 1024 PAGE 5
`
`

`

`5,635,517
`
`10
`talc, magnesium stearate and half of the starch are intimately
`mixed. The other half of the starch is suspended in 65 mL of
`water and this suspension is added to a boiling solution of
`the polyethylene glycol in 260 mL of water. The resulting
`5 paste is added to the pulverulent substances, and the whole
`is mixed and granulated, if necessary with the addition of
`water. The granulate is dried overnight at 35° C., forced
`through a sieve of 1.2 mm mesh width and compressed to
`10 form tablets of approximately 10 mm diameter which are
`concave on both sides and have a breaking notch on the
`upper side.
`
`15
`
`20
`
`25
`
`EXAMPLE?
`
`Gelatin dry-filled capsules, each containing 100 mg of
`1-oxo-2-(2,6-dioxopiperidin-3-yl)-6-arninoisoindoline, can
`be prepared in the following manner:
`
`Composition (for 1000 capsules)
`
`1-oxo-2-(2,6-dioxo(cid:173)
`piperidin-3-yl}-6-amino(cid:173)
`isoincloline
`microcrystalline cellulose
`sodium lauryl sulfate
`magnesium stearate
`
`100.0 g
`
`30.0g
`2.0g
`8.0 g
`
`9
`-continued
`
`Constituents (fur 1000 tablets)
`
`lactose
`wheat stan:h
`magnesium stearate
`
`100.0 g
`47.0g
`3.0 g
`
`All the solid ingredients are first forced through a sieve of
`0.6 mm mesh width. The active ingredient, lactose, magne(cid:173)
`sium stearate and half of the starch then are mixed. The other
`half of the starch is suspended in 40 mL of water and this
`suspension is added to 100 mL of boiling water. The
`resulting paste is added to the pulverulent substances and the
`mixture is granulated, if necessary with the addition of
`water. The granulate is dried overnight at 35° C., forced
`through a sieve of 1.2 mm mesh width and compressed to
`form tablets of approximately 6 mm diameter which are
`concave on both sides.
`
`EXAMPLES
`Tablets for chewing, each containing 75 mg of 1-oxo-2-
`(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, can be pre(cid:173)
`pared in the following manner:
`
`Composition (for 1000 tablets)
`
`1-oxo-2-(2,6-dioxo(cid:173)
`pipcridin-3-yl}-4-amino(cid:173)
`isoincloline
`mannitol
`lactose
`talc
`glycine
`stcaric acid
`saccharin
`5% gelatin solution
`
`75.0 g
`
`230.0 g
`150.0 g
`21.0 g
`12.5 g
`10.0g
`1.5 g
`q.s.
`
`30
`
`The sodium lauryl sulfate is sieved into the 1-oxo-2-(2,
`6-dioxopiperidin-3-yl)-6-aminoisoindoline through a sieve
`of 0.2 mm mesh width and the two components are inti(cid:173)
`mately mixed for 10 minutes. The rnicrocrystalline cellulose
`35 is then added through a sieve of 0.9 mm me