United States Patent 1191
`Nelson et a1.
`
`[11] Patent Number:
`[45] Date of Patent:
`
`4,952,579
`Aug. 28, 1990
`
`[54] METHOD OF TREATING DISEASES BY
`ADMINISTERING
`MORPHOLINO-ETHYLESTER OF
`MY COPHENOLIC ACID OR DERIVATIVES
`THEREOF
`[75] Inventors: Peter H. Nelson, Los Altos;
`Chee-Liang L. Gu, Synnyvale;
`Anthony C. Allison; Elsie M. Eugui,
`both of Belmont; William A. Lee,
`Menlo Park, all of Calif.
`[73] Assignee:
`Syntex (U.S.A.) Inc., Palo Alto, Calif.
`[21] Appl. No.: 272,161
`[22] Filed:
`Nov. 14, 1988
`
`[62]
`
`Related US. Application Data
`Division of Ser. No. 93,459, Sep. 4, 1937, Pat. No.
`4,808,592, which is a division of S61‘. No. 8,717, Jan. 30,
`1937, Pat. No. 4,753,935.
`
`[51] Im. (11.5 ................. .. A61K 31/535; com 413/12
`[52] US. 01. ................................ .. 514/233.5; 544/153
`[53] Field of Search .................... .,. 544/153; 514/233.5
`
`[56] >
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,705,894 12/ 1972 Gerzon et a1. ................ .. 260/240 R
`
`3,777,020 12/ 1973 Johnson . . . . . . .
`
`. . . . . .. 424/180
`
`260/3433
`3,825,571 7/1974 Mori et al.
`260/3433
`3,853,919 12/1974 Mori et al.
`424/248
`3,868,454 2/1975 Johnson ..... ..
`424/180
`3,880,995 4/1975 Jones ...... ..
`3,903,071 9/1975 Holmes ......................... .. 260/210 R
`
`FOREIGN PATENT DOCUMENTS
`
`86860 11/1973 Japan .
`86861 ll/1973 Japan .
`57-024380 2/1982 Japan .
`57-183776 ll/1982 Japan .
`57-183777 1l/l982 Japan .
`
`1157100 7/1969 United Kingdom .
`1261060 l/1972 United Kingdom .
`2117238 10/1983 United Kingdom .
`OTHER PUBLICATIONS
`“Antitumor Activity of Derivatives of Mycophenolic
`Acid”, Suzuki, et al., J. Antibiotics, 29(3), 275-285,
`1975.
`“Carbamoyl Mycophenolic Acid Ethylester, An Oral
`Antitumor Agen ”, Suzuki, et al., J. Antibiotics, 29(3),
`286-291, 1975.
`“Antitumor and Immunosuppressive Effects of Myco
`phenolic Acid”, Ohsugi, et al., Cancer Research, 36,
`2923—2927, 1976.
`“A Gastroprotective Anti-In?ammatory Agent: The
`B-Morpholinoethyl Ester of Niflumic Acid (Morni?u
`mate)”, Schiantarelli, et al., Agents and Actions, 14(2),
`1984.
`Primary Examiner—.lohn M. Ford
`Attorney. Agent, or Firm—David A. Lowin; Tom M.
`Moran
`ABSTRACT
`[57]
`The compounds and pharmaceutical compositions of
`Formula A, wherein Z is hydrogen or —-C(O)R, where
`R is lower alkyl or aryl, and the pharmaceutically ac
`ceptable salts thereof, are useful as immunosuppressive
`agents, anti-in?ammatory agents, anti-tumor agents,
`anti-viral agents, and anti-psoriatic agents.
`
`0
`\
`
`0
`
`0- 2
`
`CH3
`
`(Formula A)
`
`’
`
`c—0—c112—c112—N
`
`o
`
`0cm
`
`CH3
`
`8 Claims, No Drawings
`
`NOVARTIS EXHIBIT 2026
`Par v Novartis, IPR 2016-00084
`Page 1 of 10
`
`

`
`1
`
`IWEI'HOD OF TREATING DISEASES BY
`ADMINISTERING MORPHOLINO-ETHYIES'I'ER
`OF MYCOPHENOLIC ACID OR DERIVATIVES
`THEREOF
`
`4,952,579
`2
`and the pharmaceutically acceptable salts thereof,
`where:
`R1 is H or lower alkyl having 1 to 6 carbon atoms;
`R; is H, lower alkyl having 1 to 6 carbon atoms or
`~phenyl-4-CO2R3, in which R3 is H, lower alkyl having
`1 to 6 carbon atoms or a pharmaceutically acceptable
`cation;
`R4 and R5 are each independently H or lower alkyl
`having 1 to 6 carbon atoms;
`X1 and Y1 are each independently O or S; and
`q is an integer of 1-6.
`Ser. No. 821,633, ?led Jan. 23, 1986, relates to com
`pounds having the general structure of Formula 2:
`
`AR1
`
`(Formula 2)
`
`OCH3
`
`CH3
`
`and the pharmaceutically acceptable salts thereof,
`where:
`A is oxygen or sulfur;
`R1 is selected from the group consisting of:
`
`A1
`I
`
`0
`ll
`
`Q H
`
`A1 is oxygen or sulfur;
`q is an integer from 0-6;
`R2 is alkyl, haloalkyl or —NR4Rs, where: R4 and R5
`are independently H, alkyl, haloalkyl, cycloalkyl,
`phenyl optionally monosubstituted with halogen,
`hydroxy, carboxy, chlorocarbonyl, sulfonylamino,
`nitro, cyano, phenyl, alkyl, acyl, alkoxycarbonyl,
`acylamino, dialkylamino or dialkylaminoethox
`ycarbonyl, phenyl optionally disubstituted with
`hydroxy, carboxy, nitro or alkyl, or benzyl option
`ally substituted with dialkylamino;
`R3 is H, alkyl or a pharmaceutically acceptable cat
`ion;
`Q and Q1 are independently H or —CO2R3; and
`Z1 is selected from the group consisting of: IH-tet
`razolyl, —CHZOH, —CHO, —CN, --C(O)A2R6 and
`—-C(O)NR7Rg, in which:
`A; is oxygen or sulfur;
`R6 is H, alkyl, alkenyl, cycloalkyl, optionally substi
`tuted phenyl, optionally substituted benzyl or a
`pharmaceutically acceptable cation; and
`R7 and R3 are independently H, alkyl or cycloalkyl,
`or R7 and R8 taken together are ——(CH2)2O(CH2.
`)2—-, —(CH2)4—. 0r —(CH2)s—;
`with the proviso that R; and R6 cannot both be H if A
`and A2 are oxygen.
`Compounds somewhat structurally similar to the
`compounds of Formulae l and 2 are described in U.S.
`Pat. Nos. 3,705,894; 3,853,919; 3,868,454; 3,880,995, in
`Japanese Pat. No. I 57024380, in J. Antibiot, 29(3),
`275-85, 286-91 (1976), and in Cancer Research, 36(8),
`2923-7 (1976). The disclosed compounds are described
`as having anti-tumor, immunosuppressive, anti-viral,
`anti-arthritic and/or anti-psoriastic activities.
`
`This is a division of pending application Ser. No.
`093,459, now U.S. Pat. No. 4,808,592 ?led Sept. 4, 1987,
`which in turn is a division of application Ser. No.
`(X)8,717 ?led Jan. 30, 1987, now U.S. Pat. No. 4,753,935,
`issued June 28, 1988, incorporated herein by reference.
`BACKGROUND OF THE INVENTION
`1. Field of the Invention
`The present invention relates to pharmaceutical com
`positions, particularly to the morpholinoethyl ester of
`mycophenolic acid and certain simple ester derivatives
`of the phenolic hydroxyl group, and to their use as
`immunosuppressive and anti-in?ammatory agents. For
`example, they are useful for treating rheumatoid arthri
`tis, in which there is an immunologically driven in?am
`matory process. Because of their effects on purine me
`tabolism, the pharmaceutical compositions of the pres
`ent invention also ?nd use as anti-tumor, anti-viral and
`anti-psoriatic agents.
`2. Cross-Reference to Related Applications
`This application is related to Ser. No. 008,909 now
`U.S. Pat. No. 4,727,069, entitled “Heterocyclic Amino
`alkyl Esters of Mycophenolic Acid and Derivatives
`Thereo ,” ?led Jan. 30, 1987 to Ser. No. 803,041, ?led
`Nov. 27, 1985; and to Ser. No. 821,633, ?led Jan. 23,
`1986.
`
`25
`
`30
`
`BACKGROUND INFORMATION AND
`RELATED DISCLOSURES
`In?ammatory diseases, in particular rheumatoid ar
`thritis, have been treated with a variety of compounds
`representing several structural classes and biological
`activities, including, for example, anti-in?ammatory
`agents (corticosteroids, aspirin, derivatives of arylacetic
`and arylpropionic acids, and oxicams), immunosuppres
`sive agents and regimes (methotrexate, cyclophospha
`mide, cyclosporin, and total lymphoid irradiation), and
`long-acting anti-rheumatic drugs (gold salts, and peni
`cillamine and its derivatives). However, no representa
`tive of any of these classes of compounds is regarded as
`ideal.
`Mycophenolic acid is a weakly-active antibiotic
`found in the fermentation broth of Penicillium brevicom
`pactum. Some compounds relating to mycophenolic
`acid, and their uses in the treatment of in?ammatory
`diseases, such as rheumatoid arthritis, are disclosed in
`the following two prior related applications.
`Ser. No. 803,041, ?led Nov. 27, 1985, relates to com
`pounds having the general structure of Formula 1:
`
`55
`
`0
`\\
`
`0
`
`Y a
`u‘ / ‘
`OCN
`\
`R2
`
`Gem
`
`CH3
`
`(Formula 1)
`
`1')
`
`X1(CH2)q7—\
`o
`0
`R4><R5
`
`NOVARTIS EXHIBIT 2026
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`Page 2 of 10
`
`

`
`3
`SUMMARY OF THE INVENTION
`One aspect of the present invention concerns the
`morpholinoethyl ester of mycophenolic acid and cer
`tain derivatives of mycophenolic acid, i.e., compounds
`having the structure of Formula A, which follows:
`
`0
`\\
`0
`
`0-2
`
`CH
`’
`
`OCI'I3
`
`CH3
`
`(Formula A)
`/"'\
`C-O-CHz-CI-Iz-N
`O
`
`wherein Z is hydrogen or —C(O)R, where R is lower
`alkyl or aryl, and the pharmaceutically acceptable salts
`thereof.
`In mother aspect, the invention relates to a pharma
`20
`ceutical composition containing a therapeutically effec
`tive amount of a compound of Formula A admixed with
`at least one pharmaceutically acceptable excipient.
`In still another aspect, the invention relates to a
`method of treating autoimmune disorders, psoriasis,
`in?ammatory diseases including in particular rheuma
`toid arthritis, and for treating tumors and viruses in a
`mammalbyadministeringtoamammalinneedofsuch
`treatment a therapeutically e?ective amount of a com
`pound of Formula A.
`Compounds of Formula A have advantageous phar
`macokinetic properties, for example, solubility in the
`delivery environment (e.g., the stomach), peak plasma
`concentration, maximum plasma concentration, and
`improved activity, e.g., anti-in?ammatory activity as
`compared to mycophenolic acid.
`DETAILED DESCRIPTION OF THE
`INVENTION
`De?nitions and General Parameters
`The following de?nitions are set forth to illustrate
`and de?ne the meaning and scope of the various terms
`used to describe the invention herein.
`The numbering of the mycophenolic acid is as fol
`lows:
`
`35
`
`4,952,579
`4
`(l,3-dihydro-4-benzoyloxy~6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl-4-hexenoate.”
`As used herein, the term “alkyl" refers to a fully
`saturated monovalent radical containing only carbon
`and hydrogen, and which may be a cyclic, branched or
`straight chain radical. This term is further exempli?ed
`by radicals such as methyl, ethyl, t-butyl, pentyl, heptyl
`and pivalyl.
`The term “lower alkyl” refers to a monovalent alkyl
`radical of one to six carbon atoms. This term is further
`exempli?ed by such radicals as methyl, ethyl, n-propyl,
`isopropyl, n-butyl, t-butyl, i-butyl (or Z-methylpropyl),
`isoamyl, pentyl, and isopentyl.
`The term “ary ” refers to a substituted or unsubsti
`tuted monovalent unsaturated aromatic carbocylic radi
`cal having a single ring (e.g., phenyl) or two condensed
`rings (e.g., naphthyl).
`The term “acy ” refers to a radical based on an or
`ganic acid, e.g., —C(O)Rl where R1 is alkyl or aryl.
`As used herein, the term “halo" refers to ?uoro,
`bromo, chloro and iodo.
`Isolation and puri?cation of the compounds and in
`termediates described herein can be e?'ected, if desired,
`by any suitable separation or puri?cation procedure
`such as, for example, ?ltration, extraction, crystalliza
`tion, column chromatography, thin-layer chromatogra
`phy or thick-layer chromatography, or a combination
`of these procedures. Speci?c illustrations of suitable
`separation and isolation procedures can be found by
`reference to the examples hereinbelow. However, other
`equivalent separation or isolation procedures can, of
`course, also be used.
`A “pharmaceutically acceptable salt” may be any salt
`derived from an inorganic or organic acid. The term
`“pharmaceutically acceptable anion” refers to the anion
`of such salts. The salt and the anion are chosen not to be
`biologically or otherwise undesirable. These salts are
`formed with inorganic acids such as hydrochloric acid,
`hydrobromic acid, sulfuric acid (giving the sulfate and
`bisulfate salts), nitric acid, phosphoric acid and the like,
`and organic acids such as acetic acid, propionic acid,
`glycolic acid, pyruvic acid, oxalic acid, malic acid,
`malonic acid, succinic acid, maleic acid, fumaric acid,
`tartaric acid, citric acid, benzoic acid, cinnamic acid,
`mandelic acid, methanesulfonic acid, ethanesulfonic
`acid, p-toluenesulfonic acid, salicylic acid and the like.
`As used herein, the term “treatmen ” or “treating”
`means any treatment of a d'nease in a mammal, and
`includes:
`(i) preventing the disease, that is, causing the clinical
`symptoms of the disease not to develop;
`(ii) inhibiting the diseae, that is, arresting the devel
`opment of clinical symptoms; and/or
`(iii) relieving the disease, that is, causing the regres
`sion of clinical symptoms.
`Unless speci?ed to the contrary, the reactions de
`scribed herein take place at atmospheric pressure over a
`temperature range from about 10' C. to about 100' C.,
`more preferably from about 10‘ C. to about 50' C., and
`most preferably at about room temperature.
`Preparation of the Compounds of Formula A
`The compounds of Formula A can be prepared ac
`cording to several synthetic pathways, depending upon
`the substitution at Z, typically starting with mycophe
`nolic acid, which is commercially available. Where Z is
`—C(O)R, the phenolic oxygen or mycophenolic acid
`can be acylated either before or a?er the esteri?cation
`
`45
`
`CH3
`
`The compounds of the invention will be named using
`the above-shown numbering system as the morpholino
`ethyl esters of E-6-(l,3-dihydro-4-hydroxy-6-methoxy
`7-methyl-3-oxo-5-isobenzofuranyl)-4-methyl-4-hex
`enoic acid and its derivatives. The compounds of the
`present invention are prepared as the E (or Entgegen)
`position isomer. Some representative compounds are
`named as follows:
`the compound of Formula A where Z is —-C(O)R and
`wherein R is methyl is named “morpholinoethyl E-6
`(1,3-dihydro-4-acetoxy-6-methoxy-7-methyl-3-oxo-5
`isobenzo?iranyl)-4-methyl-4-hexenoate” and
`the compound of Formula A where Z is —C(O)R and
`wherein R is phenyl is named “morpholinoethyl E6
`
`60
`
`NOVARTIS EXHIBIT 2026
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`
`

`
`4,952,579
`5
`6
`of the acid. Where Z is hydrogen, the starting material
`about 3 molar equivalents, of an inorganic base (such as
`is typically mycophenolic acid.
`sodium carbonate, potassium bicarbonate or the like) or
`a tertiary organic base (such as triethylamine, N
`Morpholinoethyl Esteri?cation of Mycophenolic Acids
`methylpiperidine or preferably pyridine). Certain bases
`Many standard esteri?cation procedures may be
`(e.g., pyridine) can also serve as the inert organic sol
`used, for example, as described in Synthetic Organic
`vent. The reaction takes place at a temperature of about
`Chemista by R. B. Wagner and H. D. Zook (Wiley,
`0'-25' 0., preferably about 5° C., for about l-lO hours,
`New York) 1956, see pages 479-532. Two presently
`preferably about 3 hours. When the reaction is substan
`tially complete, the acylated product is isolated by con
`preferred synthetic routes are described below for con
`version of mycophenolic acid and its derivatives into
`ventional means.
`the morpholinoethyl ester compounds of Formula A.
`The ?rst route involves conversion into an acid halide,
`followed by condensation with morpholinoethanol to
`the end product. The second route involves conversion
`directly into the end product using a carbodiimide reac
`tion.
`As an example, a less preferred third route entails
`‘starting with an ester of mycophenolic acid (other than
`the morpholinoethyl ester) in an ester exchange reac
`tion for conversion into the desired end product.
`
`20
`
`15
`
`Salts of Compounds of Formula A
`The compounds of Formula A may be converted to
`corresponding acid addition salts. The conversion is
`accomplished by treatment with at least a stoichiomet
`ric amount of an appropriate acid, such as hydrochloric
`acid, sulfuric acid, methanesulfonic acid or the like.
`Typically, the free base is dissolved in a polar organic
`solvent such as ethanol, methanol, or ethyl acetate and
`the acid added in water, ethanol, methanol, or isopropa
`nol. The temperature is maintained at 0'-50' C. The
`resulting salt precipitates spontaneously or may be
`brought out of solution with a less polar solvent.
`A dibasic acid, such as sulfuric acid, can form two
`salts with the compounds of this invention. One such
`salt, in which one mole of the base and one mole of the
`acid are present, is called the bisulfate (or hydrogen
`sulfate) salt. The other, in which two moles of the base
`and one mole of the acid are present, is called the sul
`fate.
`The acid addition salts of the compounds of Formula
`A may be decomposed to the corresponding free bases
`by treating with an excess of a suitable base, such as
`ammonia or sodium bicarbonate, typically in the pres
`ence of aqueous solvent, and at a temperature of be
`tween O' and 50’ C. The free base form is isolated by
`conventional means, such as extraction with an organic
`solvent.
`
`Preferred Compounds
`Most preferred are the compound of Formula A
`where Z is hydrogen, i.e., morpholinoethyl E-6-(l,3
`dihydro-4-hydroxy-6-methoxy-7—methyl-3-oxo-5
`isobenzofuranyl)-4-methyl-4-hexenoate, and its pharma
`ceutically acceptable salts (preferably the hydrochlo
`ride, sulfate and bisulfate salts).
`Also preferred are the following compounds and
`pharmaceutically acceptable salts (preferable the hy
`drochloride, sulfate and bisulfate salts) of Formula A
`where Z is —C(O)R:
`E-6-(1,3-dihydro-4-acetoxy-6
`morpholinoethyl
`methoxy-7-methyl-3—oxo-5-isobenzofuranyD-4-meth
`yl-4-hexenoate;
`morpholinoethyl E-6-(l,3-dihydro-4-propionyloxy-6
`methoxy-7-methyl-3-oxo-5-isobenzofuranyD-4-meth
`yl-4-hexenoate;
`morpholinoethyl
`E-6-(l,3-dihydro~4-pivaloyloxy-6
`methoxy-7-methyl-3-oxo-5-isobenzofuranyly4-meth
`yl-4-hexenoate; and
`morpholinoethyl
`E-6-(l,3-dihydro-4-benzoyloxy-6
`methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-meth
`yl-4-hexenoate.
`Preferred Processes
`The compounds of the present invention can be pre
`pared according to the following last steps:
`an E-6-(1,3—dihydro—4-hydroxy-6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl—4-hexenoyl halide,
`
`25
`
`30
`
`35
`
`The Acid Halide-Condensation Route
`In the ?rst synthetic route, mycophenolic acid or an
`acylated derivative thereof is dissolved or suspended in
`a solvent inert under the conditions of the reaction (i.e.,
`an inert solvent, such as benzene, toluene, acetonitrile,
`tetrahydrofuran, diethyl ether, chloroform or prefera
`bly methylene chloride) and an excess (about 10 molar
`equivalents to l) of a halogenating agent (e.g., thionyl
`chloride) is added, optionally together with a small
`amount of dimethylformamide. The reaction mixture is
`stirred for about 1-8 hours, preferably about 4 hours, to
`yield the corresponding acid halide.
`The acid halide is dissolved in an inert solvent, as
`described above, and reacted by a condensation reac
`tion with a cooled solution (e.g., maintained at about 40'
`C.) of morpholinoethanol [also named as 4-(2-hydroxye
`thyl)-morpholine], to which it is added slowly over a
`period of about 10 minutes to 2 hours, preferably about
`90 minutes. The end product of Formula A is isolated
`and puri?ed by conventional procedures.
`The Carbodiimide Route
`In the second synthetic route, mycophenolic acid or
`an acylated derivative thereof is dissolved in a solvent
`inert under the conditions of the reaction [such as dry
`tetrahydrofuran (“THF”), dichloromethane, or carbon
`tetrachloride; preferably THF] and reacted with mor
`pholinoethanol in the presence of a carbodiimide, such
`as DCC (“dicyclohexylcarbodiimide”) or di-p-tolylcar
`bodiimide. The molar ratio of alcohol to the starting
`acid is about 1:1. The reaction takes place at atmo
`spheric pressure over a period of about 4-8 hours, pref
`erably over 6 hours. A temperature range from about
`10' C. to about reflux temperature, preferably about
`55
`room temperature may be used. The end product of
`Formula A is isolated and puri?ed in the usual manner.
`Acylation of the Phenolic Oxygen
`The compounds of Formula A where Z is —C(O)R
`are prepared by dissolving mycophenolic acid or the
`morpholinoethyl ester thereof in an inert organic sol
`vent as defined above (e.g., acetonitrile or preferably
`pyridine) and reacting it with about 1 to 6 molar equiva
`lents, preferably about 3 molar equivalents, of the ap
`propriate acyl halide or anhydride (e.g., acetic anhy
`dride, propionyl chloride or pivaloyl chloride) in the
`presence of about 1 to 6 molar equivalents, preferably
`
`45
`
`50
`
`65
`
`NOVARTIS EXHIBIT 2026
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`
`

`
`5
`
`8
`described by Greaves, et al. [“Activation of human T
`and B lymphocytes by polyclonalmitogens,” Nature,
`248, 698-701 (1974)].
`Anti-viral activity is determined by the procedure
`described by Smee, et al. [“Anti-Herpesvirus Activity
`of the Acyclic Nucleoside 9-(l,3-Dihydroxy-2-Propox
`ymethyDGuanine,” Antimicrobial Agents and Chemo
`therapy, 23 (5), 676-682 (1983)] or as described by
`Planterose [“Antiviral and cytotoxic effects of myco
`phenolic acid,” Journal of General Virology, 4, 629
`
`zonal-min,
`
`,
`
`.
`
`45
`
`UTILITY AND ADMINISTRATION
`
`4,952,579
`7
`is condensed with morpholinoethanol to give a com
`pound according to Formula A where Z is hydrogen;
`an E-6-(l,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl-4-hexenoic acid is
`contacted with morpholinoethanol in the presence of
`a. carbodiimide to give a compound according to
`Formula A where Z is hydrogen;
`an E-6(l,3dihydro-4-acyloxy-6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl-4~hexenoyl halide,
`is condensed with morpholinoethanol to give a com
`pound according to Formula A where Z is —C(O)R;
`an E-6-(l,3-dihydro-4-acyloxy-6-methoxy-7-methyl-3
`_Tests for systemic activity in psoriasis can be carried
`oxo-5-isobenzofuranyl)-4-methyl-4-hexenoic acid is
`out as described by Spatz, et al. [“Mycophenolic acid in
`condensed with morpholinoethanol in the presence of
`psoriasis,” British Journal of Dermatology, 98, 429
`a carbodiimide to give a compound according to
`(1978)].
`‘ Formula A where Z is —C(O)R;
`Tests for anti-tumor activity can be performed as
`morpholinoethyl
`E-6-(1,3-dihydro-4-hydroxy-6
`described by Carter, et al. [“Mycophenolic acid: an
`methoxy-7-methyl-3-oxo-5-isobenzo?1ranyD-4-meth
`anti-cancer compound with unusual properties,” Na
`yl-4-hexenoate is condensed with an acyl halide or
`ture, 223, 848 (1969)].
`anhydride to give a compound according to Formula 20
`A where Z is —C(O)R;.
`General Administration
`contacting a pharmaceutically acceptable acid with a
`Administration of the active compounds of Formula
`compound of Formula A to form the corresponding
`A, in pure form or in an appropriate pharmaceutical
`acid addition salt of Formula A;
`composition can be carried out via any of the accepted
`substituting a pharmaceutically acceptable acid salt of 25
`modes of administration of agents for serving similar
`Formula A with another pharmaceutically accept
`utilities. Thus, administration can be, for example,
`able acid; and
`orally, nasally, parenterally or topically, in the form of
`contacting an acid addition salt of Formula A with a
`solid, semi-solid, lyophilized powder, or liquid dosage
`base to form the corresponding free base compounds
`forms such as for example, tablets, suppositories, pills,
`of Formula A.
`30
`capsules, powders, solutions, suspensions, emulsions,
`creams, lotions, aerosols, ointments or the like, prefera
`bly in unit dosage forms suitable for simple administra
`tion of precise dosages. The compositions will include a
`conventional pharmaceutical carrier or excipient and an
`active compound of Formula A and, in addition, may
`include other medicinal agents, pharmaceutical agents,
`carriers, adjuvants, etc.
`Generally, depending on the intended mode of ad
`ministration, the pharmaceutically acceptable composi
`tions will contain about 1% to about 99% by weight of
`the pharmaceutically active compound of this invention
`and 99% to 1% by weight of suitable pharmaceutical
`excipients. Preferably, the composition will be about 5
`to 75% by weight of a pharmaceutically active com
`pound, with the rest being suitable pharmaceutical ex
`cipients.
`The preferred manner of administration, for the con
`ditions detailed above, is oral using a convenient daily
`dosage regimen which can be adjusted according to the
`degree of affliction. For such oral administration, a
`pharmaceutically acceptable, non-toxic composition is
`formed by the incorporation of any of the normally
`employed excipients, such as, for example, pharmaceu
`tical grades of mannitol, lactose, starch, magnesium
`stearate, sodium saccharine, talcum, cellulose, glucose,
`gelatin, sucrose, magnesium carbonate, and the like.
`Such compositions take the form of solutions, suspen
`sions, tablets, pills, capsules, powders, sustained release
`formulations and the like.
`Preferably the compositions will take the form of a
`pill or tablet and thus the composition will contain,
`along with the active ingredient, a diluent such as lac
`tose, sucrose, dicalcium phosphate, and the like; and a
`binder such as a starch, gum acacia, polyvinylpyrroli
`done, gelatin, cellulose and derivatives thereof‘, and the
`like.
`The active compounds of Formulas I may be formu
`lated into a suppository using, for example, about 0.5%
`
`General Utility
`The compounds of the present invention, including 35
`the pharmaceutically acceptable salts thereof, and the:
`compositions containing them, are useful as immuno
`suppressive agents, anti-inflammatory agents, anti
`tumor agents, anti-viral agents, and anti-psoriatic agents
`in mammals, whether domestic (cattle, pigs, sheep, 40
`goats, horses), pets (cats, dogs), 0r preferably humans.
`For example compounds of Formula A are useful for
`treating rheumatoid arthritis, in which there is an immu-
`nologically driven in?ammatory process. These com
`pounds can be used both prophylactically (e.g., to pre
`vent allogra? rejection) and therapeutically.
`Testing
`Initial animal screening tests to determine anti-in?am
`matory activity potential include the adjuvant arthritis
`assay according to the method of Pearson, Proc. Soc.
`Exp. Biol. Med., 91:95-10] (1956).
`Also, in vitro tests, for example those using synovial
`explants from patients with rheumatoid arthritis, Dayer,
`et al., J. Exp. Med, 145:1399-1404 (1977), are useful in
`determining whether compounds exhibit anti-in?amma
`tory activity.
`Autoimmune activity is determine utilizing experi
`mental allergic encephalomyelitis by a modi?cation of a
`procedure initially described by Grieg, et al., J. Phar
`macol. Exp. Ther. 173:85 (1970).
`‘
`Immunosuppressive activity is determined by both in
`vivo and in vitro procedures. In vivo activity is deter
`mined utilizing a modi?cation of the Jerne hemolytic
`plaque assay, [.lerne, et al., “The agar plaque technique
`for recognizing antibody producing cells,” Cell-bound
`Antibodies, Amos, B. and Kaprowski, H. editors (Wistar
`Institute Press, Philadelphia) 1963, p. 109]. In vitro
`activity is determined by an adaptation of the procedure
`
`50
`
`55
`
`60
`
`65
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`NOVARTIS EXHIBIT 2026
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`4,952,579
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`9
`to about 50% active ingredient disposed in a carrier of
`aqueous sodium bicarbonate. The organic solution was
`polyethylene glycols (PEG) [e.g., PEG 1000 (96%) and
`dried with sodium sulphate and evaporated to yield
`morpholinoethyl
`E-6-(l,3-dihydro-b
`4-hydroxy-6
`PEG 40(1) (4%)].
`methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-methyl
`Liquid pharmaceutically administerable composi
`tions can, for example, be prepared by dissolving, dis
`4-hexenoate (m.p. 93'—94' C.
`persing, etc. an active compound (about 0.5% to about
`20%), as described above, the optional pharmaceuti
`cally adjuvanis in a carrier, such as, for example, water,
`saline, aqueous dextrose, glycerol, ethanol and the like,
`to thereby form a solution or suspension.
`If desired, the pharmaceutical composition to be ad
`ministered may also contain minor amounts of non-toxic
`auxiliary substances such as wetting or emulsifying
`agents, pH buffering agents and the like, such as for
`example, sodium acetate, sorbitan monolaurate, trietha
`nolamine oleate, etc.
`Actual methods of preparing such dosage forms are
`known, or will be apparent, to those skilled in this art;
`for example, see Remington’s Pharmaceutical Sciences,
`16th Ed., (Mack Publishing Company, Easton, Pa.,
`1980). The composition to be administered will, in any
`event, contain a quantity of the active compound(s) in a
`pharmaceutically effective amount for relief of the par
`ticular condition being treated when administered in
`accordance with the teachings of this invention.
`Generally, the compounds of the invention are ad
`ministered in a therapeutically effective amount, i.e., a
`dosage sufficient to effect treatment, which will vary '
`depending on the individual and condition being
`treated. Typically, a therapeutically e?‘ective daily dose
`is from 0.02 to 100 mg/kg of body weight per day of an
`active compound of Formula A. Most conditions re
`spond to treatment comprising a dosage level on the
`order of 0.4 to 30 mg/kg of body weight per day, and
`most preferably about 10 mg/kg/day. Thus, for admin
`istration to a 70 kg person, the dosage range would be
`about 1.4 mg to 7 g per day, preferably about 7.0 to 700
`mg per day.
`
`20
`
`25
`
`EXAMPLE 2
`Morpholinoethyl
`E-6-(l,3-dihydro-4-acetoxy-6-methoxy-7-methyl-3-oxo
`5-isobenzofuranyl)-4-methyl-4-hexenoate
`2A. Formula A where Z is —C(O)CH3
`Morpholinoethyl
`E-6-(l,3-dihydro-4-hydroxy-6
`methoxy-7methyl-3-oxo-5-isobenzofuranyl)-4-methyl
`4-hexenoate (10.0 g) was dissolved in pyridine (50.0 ml)
`followed by the addition of acetic anhydride (10.0 ml).
`The mixture was stirred at room temperature for 90
`minutes, then poured into water and extracted with
`ethyl acetate. The organic solution was dried and evap=
`orated to give morpholinoethyl E-6-(l,3-dihydro-4
`acetoxy-6-methoxy-7-methyl-3-oxo-5-isobenzofuranyl)
`4-methyl-4-hexenoate.
`2B. Formula A where Z is Other Than —C(O)CH3
`Similarly, by following the procedure of part A
`above and substituting for acetic anhydride the follow
`ing materials:
`propionyl chloride,
`Z-methylpropionyl chloride,
`pivaloyl chloride, and
`benzoyl bromide;
`there are obtained the following respective compounds:
`morpholinoethyl E-6-(1,3-dihydro-4-propionyloxy-6
`methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-meth
`yl-4-hexenoate,
`E-6-[l,3-dihydro-4~(2-methylpro
`morpholinoethyl
`pionyloxy)-6-methoxy-7-methyl-3-oxo-5-isoben
`zofuranyl]-4-methyl-4-hexenoate,
`morpholinoethyl
`E-6-(l,3-dihydro-4-pivaloyloxy-6~
`methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-meth
`yl-4-hexenoate, and
`morpholinoethyl
`E-6-(l,3-dihydro-4-benzoyloxy-6
`methoxy-7-methyl-3-oxo~5-isobenzofuranyl)-4-meth
`yl-4-hexenoate.
`‘
`
`EXAMPLE 3
`Morpholinoethyl
`E-6-(l,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl-4-hexenoate
`hydrochloride
`3A. Hydrochloride Salt of Formula A where Z is
`Hydrogen
`E-6-(l,3-dihydro-4-hydroxy-6
`Morpholinoethyl
`methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-methyl
`4-hexenoate (38.0 g) was dissolved in isopropanol (200
`ml) and the solution was added to a solution of hydro
`gen chloride (l0.0 g) in isopropanol (150 ml). The hy
`drochloride salt was collected by ?ltration and dried
`under vacuum (m.p. l54°-l55' C.).
`
`3B. Hydrochloride Salts of Formula A where Z is
`Other Than —C(O)CH3
`Similarly, by following the procedure of part A
`above and substituting for morpholinoethyl E-6-(l,3
`dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxo-5
`isobenzofuranyl)-4-methyl-4-hexenoate the following
`materials (prepared, e.g., as in Example 2B):
`
`EXAMPLES
`The following examples are given to enable those
`skilled in the art to more clearly understand and to
`practice the present invention. They should not be con
`sidered as a limitation on the scope of the invention, but
`merely as being illustrative and representative thereof.
`
`45
`
`EXAMPLE 1
`Morpholinoethyl
`E-6-(l,3-dihydro-4-hydroxy-6
`methoxy-7-methyl-3-oxo-5-isoenzofuranyl-4-methyl-4
`hexenoate
`
`lA. Formula A where Z is Hydrogen
`E6-(l,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3
`oxo-5-isobenzofuranyl)-4-methyl-4-hexenoic acid (my
`cophenolic acid) (32.0 g) was dissolved in dichloro
`methane (250 ), followed by the addition of thionyl
`chloride (25.0 ml) and dimethylformamide (0.3 ml). The
`reaction mixture was stirred at room temperature for 3
`hours, after which the volatile components were re
`moved under vacuum to afford E-6-(l,3-dihydro-4
`hydroxy-6-methoxy-7-methyl-3-oxo-5-isobenzofuranl)
`4-methyl-4-hexenoic acid chloride as an oil.
`A solution of morpholinoethanol (30.5 ml) in dichlo
`romethane (250 ml) was chilled to 4' C. on an ice bath.
`The mycophenolic acid chloride oil was dissolved in
`dichloromethane (50.0 ml) and added to the chilled
`solution. After stirring for 90 minutes (at 4' C.), the
`reaction mixture was washed with water and then with
`
`NOVARTIS EXHIBIT 2026
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`morpholinoethyl E-6-(l,3-dihydro-4-propionyloxy-6
`' the active compound in the preparation of the orally
`methoxy-7-methyl-3oxo-5-isobenzo?rranyl)-4-meth
`administrable formulations of this example.
`yl-4-hexenoate,
`morpholinoethyl
`E-6-[l,3-dihydro-4-(2-